Journal of Molecular Neuroscience

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Online ISSN: 1559-1166
Print ISSN: 0895-8696
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  • Yuxiang LiaoYuxiang Liao
  • Bo LiuBo Liu
  • Chen JinChen Jin
  • [...]
  • Zhiping ZhangZhiping Zhang
In this study, we intend to identify key immune-related genes (IRGs) in gliomas using the TCGA and GTEx databases. Following collection of the RNA-seq data of lower-grade glioma (LGG) and glioblastoma (GBM) patients from the TCGA and GTEx databases, the differentially expressed IRGs (DE-IRGs) were screened. The ESTIMATE algorithm was utilized to evaluate StromalScore and ImmuneScore of LGG and GBM samples and a multifactorial Cox risk model was constructed to identify the related risk genes. The core IRGs of LGG and GBM were screened through a PPI network, followed by exploration of their correlation with glioma prognosis. The relationship between IRGs and immune cells in LGG and GBM was detected. In vitro assays were performed to detect the effect of CXCL9 on glioma cell development. We screened 403 and 492 DE-IRGs in LGG and GBM. StromalScore and ImmuneScore were related to overall survival in LGG, but not in GBM. CXCL9 was identified as a core gene in LGG and GBM and shared association with the prognosis of gliomas. Furthermore, a correlation was found between CXCL9 and immune infiltration of LGG and GBM. Glioma cell proliferation and invasion could be inhibited by silencing of CXCL9. Overall, CXCL9 is correlated to the prognosis of glioma patients and may accelerate glioma development via immune regulation.
Neuroblastoma is a childhood malignancy with high morbidity and mortality. We identified key biomarkers associated with neuroblastoma risk and prognosis. The gene modules most associated with neuroblastoma risk were derived by WGCNA. Modular genes were intersected with differentially expressed genes between patients with high-risk (HR) and non-high-risk (NHR) to obtain risk genes, and enrichment analysis was performed. After incorporating risk genes into Cox regression analysis, LASSO algorithm, and K-M survival analysis, key genes were identified and introduced into four external datasets for validation. We performed short time-series expression miner analysis and single-sample genome enrichment analysis. Finally, we evaluated the difference in DNA methylation levels to identify meaningful methylation marks. We identified 5 key genes (ANO6, CPNE2, DST, PLXNC1, SCN3A) for neuroblastoma risk and prognosis, which correlated closely with known neuroblastoma biomarkers. All key genes showed a progressive downregulation trend with increasing risk levels of neuroblastoma. The immune infiltration of 14 immune cells was significantly different between HR-NB and NHR-NB, and most immune cells were negatively correlated with key genes. Furthermore, the expression of ANO6, CPNE2, DST, and PLXNC1 was modified by DNA methylation. This study identified 5 key genes for neuroblastoma risk and prognosis that were potential biomarkers.
Focal cortical infarction leads to secondary degeneration of the ipsilateral hippocampus, which is associated with poststroke cognitive impairment. VX-765 is a potent small-molecule caspase-1 inhibitor that protects against central nervous system diseases. The present study aimed to determine the protective effects of VX-765 on β-amyloid (Aβ) deposition and secondary degeneration in the hippocampus as well as cognitive decline after cortical infarction. Sprague–Dawley rats were used to establish a distal middle cerebral artery occlusion (dMCAO) model and randomly divided into the vehicle and VX-765 groups. Rats in the vehicle and VX-765 groups, respectively, were subcutaneously injected with VX-765 (50 mg/kg/d) and an isopycnic vehicle once a day for 28 days, starting 1 h after dMCAO. At the end of this 28-day period, cognitive impairment was evaluated with the Morris water maze, and secondary hippocampal damage was evaluated with Nissl staining and immunostaining methods. Neuronal damage and pyroptosis were detected by TUNEL and immunoblotting. The results revealed that VX-765 treatment ameliorated poststroke cognitive dysfunction after ischemia. VX-765 reduced Aβ deposition, neuronal loss, and glial activation compared with the vehicle control. In addition, VX-765 treatment increased BDNF levels and normalized synaptophysin protein levels in the hippocampus after cortical infarction. Notably, VX-765 treatment significantly reduced the expression of the pyroptosis-related molecules caspase-1, NLRP3, apoptosis-associated speck-like protein (ASC), gasdermin D, IL-1β, and IL-18. Additionally, VX-765 significantly decreased the numbers of TUNEL-positive cells and the levels of Bax and cleaved caspase-3 (cC3) and enhanced the levels of Bcl-2 and Bcl-xl after ischemia. Inflammatory pathways, such as the NF-κB and mitogen-activated protein kinase (MAPK) pathways, were inhibited by VX-765 treatment after ischemia. These findings revealed that VX-765 reduced Aβ deposition, pyroptosis, and apoptosis in the ipsilateral hippocampus, which may be associated with reduced secondary degeneration and cognitive decline following focal cortical infarction.
Salvia miltiorrhiza (SAL) and Panax notoginseng (PNS) are widely used in treating of ischemic stroke. However, it is unknown which components of SAL and PNS protect brain microvascular pericytes after an ischemic stroke. We evaluated the protective effects and mechanisms of SAL and PNS components in pericytes subjected to oxygen-glucose deprivation/reoxygenation (OGD/R). Pericytes were subjected to OGD/R. Cell Counting Kit-8 (CCK-8) was used to evaluate cell viability. ROS and SOD kits were used to detect oxidative stress. Flow cytometry was performed to analyze cell apoptosis. To evaluate cell migration, a scratch assay was performed. Expression of cleaved caspase-3, Bcl-2, Bax, VEGF, Ang-1, PDGFR-β, PI3K/AKT/mTOR, and JNK/ERK/P38 signaling pathways were identified using western blot. The results revealed that salvianolic acid B (Sal B), salvianolic acid D (Sal D), notoginsenoside R1 (R1), ginsenoside Rb1 (Rb1), and ginsenoside Rg1 (Rg1) increased the cell viability of pericytes subjected to OGD/R, reduced the level of ROS, and increased the expression of SOD. The components reduced cell apoptosis, increased the protein level of Bcl-2/Bax, reduced the level of cleaved caspase-3/caspase-3, increased cell migration, and enhanced the levels of Ang-1, PDGFR-β, and VEGF. The components could activate PI3K/AKT/mTOR pathway while inhibiting the JNK/ERK/P38 pathway. Studies found that Sal B, Sal D, R1, Rb1, and Rg1 inhibited oxidative stress and apoptosis while increasing the release of pro-angiogenic regulators of pericytes related to the PI3K/AKT/mTOR and JNK/ERK/P38 signaling pathways. This provides a potential foundation for developing monomeric drugs for treating ischemic stroke.
Diagrammatic representation of proposed database correlating lnRNA, PM2.5, and glioblastoma progressions
Diagrammatic representation of pipeline of work for proposed database
Graphical illustration representing PM-mediated carcinogenic changes in glial cells potentiating glioblastoma onset and progression
Graphical summary of the study
With a reported rise in global air pollution, more than 50% of the population remains exposed to toxic air pollutants in the form of particulate matters (PMs). PMs, from various sources and of varying sizes, have a significant impact on health as long-time exposure to them has seen a correlation with various health hazards and have also been determined to be carcinogenic. In addition to disrupting known cellular pathways, PMs have also been associated with lncRNA dysregulation—a factor that increases predisposition towards the onset or progression of cancer. lncRNA dysregulation is further seen to mediate glioblastoma multiforme (GBM) progression. The vast array of information regarding cancer types including GBM and its various precursors can easily be obtained via innovative in silico approaches in the form of databases such as GEO and TCGA; however, a need to obtain selective and specific information correlating anthropogenic factors and disease progression—in the case of GBM—can serve as a critical tool to filter down and target specific PMs and lncRNAs responsible for regulating key cancer hallmarks in glioblastoma. The current review article proposes an in silico approach in the form of a database that reviews current updates on correlation of PMs with lncRNA dysregulation leading to GBM progression.
Glutamate, one of the most important excitatory neurotransmitters, acts as a signal transducer in peripheral tissues and endocrine cells. Excessive glutamate secretion has been shown to cause excitotoxicity and neurodegenerative disease. Cerebrolysin is a mixture of enzymatically treated peptides derived from pig brain including neurotrophic factors, like brain-derived neurotrophic factor (BDNF), glial cell line-derived neurotrophic factor (GDNF), nerve growth factor (NGF), and ciliary neurotrophic factor (CNTF). The present study investigated the protective effects of cerebrolysin on glutamate transporters (EAAT 1, EAAT 2) and cytokines (IL-1β and IL-10) activity in glutamate-mediated neurotoxicity. Primary cortex neuron culture was exposed to glutamate and successively treated with various cerebrolysin concentrations for 24 and 48 h. Our data showed that cerebrolysin primarily protects neurons by decreasing glutamate concentration in the synaptic cleft. In addition, Cerebrolysin can decrease oxidative stress and neuron cell damage by increasing antioxidant activity and decreasing inflammation cytokine levels.
Hypoxia is a prevalent hallmark of many malignant neoplasms. The aim was to assess the serum hypoxia biomarkers HIF-1α, VEGF, osteopontin, erythropoietin, caveolin-1, GLUT-1, and LDH pre- and post-radiotherapy in patients with brain tumors. The study was conducted on 120 subjects were divided into two groups: group I: 40 healthy volunteers as control group. Group II: 80 brain tumor patients were subdivided into glioblastoma subgroup: 40 glioblastoma patients, meningioma subgroup: 40 malignant meningioma patients. Two venous blood samples were collected from every patient prior to and following RT and one sample from controls. Biomarkers were assayed by ELISA. In glioblastoma subgroup, HIF-1α, VEGF, and LDH were significantly increased after RT. On the contrary, these biomarkers were significantly decreased after RT in malignant meningioma subgroup. Osteopontin was significantly increased after RT in both subgroups. Regarding erythropoietin, it was significantly decreased in both subgroups when compared to before RT. Caveolin-1 showed a significant increase in glioblastoma subgroup after RT comparing to before RT. GLUT-1 was significantly increased after RT in both subgroups comparing to before RT. Association of significant elevation of hypoxia biomarkers either pre- or post-RT with aggressive tumor such as glioblastoma indicates that, they are markers of malignancy and may have a role in tumor development and progression.
Coronavirus disease 2019 (COVID-19) has emerged since December 2019 and was later characterized as a pandemic by WHO, imposing a major public health threat globally. Our study aimed to identify common signatures from different biological levels to enlighten the current unclear association between COVID-19 and Parkinson's disease (PD) as a number of possible links, and hypotheses were reported in the literature. We have analyzed transcriptome data from peripheral blood mononuclear cells (PBMCs) of both COVID-19 and PD patients, resulting in a total of 81 common differentially expressed genes (DEGs). The functional enrichment analysis of common DEGs are mostly involved in the complement system, type II interferon gamma (IFNG) signaling pathway, oxidative damage, microglia pathogen phagocytosis pathway, and GABAergic synapse. The protein-protein interaction network (PPIN) construction was carried out followed by hub detection, revealing 10 hub genes (MX1, IFI27, C1QC, C1QA, IFI6, NFIX, C1S, XAF1, IFI35, and ELANE). Some of the hub genes were associated with molecular mechanisms such as Lewy bodies-induced inflammation, microglia activation, and cytokine storm. We investigated regulatory elements of hub genes at transcription factor and miRNA levels. The major transcription factors regulating hub genes are SOX2, XAF1, RUNX1, MITF, and SPI1. We propose that these events may have important roles in the onset or progression of PD. To sum up, our analysis describes possible mechanisms linking COVID-19 and PD, elucidating some unknown clues in between.
A Family pedigree. B and C Phenotypic manifestations of the proband. D, E, F, and G Shows no skin manifestations or nail dystrophy. H Umbilical hernia. I and J MRI findings. K Intraoral photo shows anterior deep overbite and normal intact healthy gingiva. L Enamel hypocalcification in upper anterior teeth, cusp tips of deciduous molars (blue arrows), and normal deciduous teeth spacing (green arrows)
Segregation analysis and localization of the detected variant in LSS protein. A and B Sanger sequencing chromatographs of the homozygous variant c.1609G > T detected in the affected female and the heterozygous carrier state of the parents. C The schematic of LSS protein structure and the positions of the novel p.Val537Leu mutation (blue) and those previously reported in the LSS gene (black) causing APMR4 syndrome
Interaction prediction analysis for p.Val537Leu using different in-silico tools. A and B The interatomic interaction of V537 with the surrounding residue in WT and of L537 in V537L mutant using Premps tool. C and D Interactions in WT and mutant residue by using DynaMut tool are colored in light-green and are also represented as sticks alongside with the surrounding residues which are involved on any type of interactions, whereas V and L represent the valine to leucine substitution at 537 position. E and F SWISS-MODEL predicted 3D for WT and mutant structures based on the X-ray structure (PDB code: 1W6K) X-ray diffraction, 2.10 Å, which contains 9 ligands: 8 × octyl beta-D-glucopyranoside and 1 × lanosterol
Secondary structure model predicted for mutant sequence by the SOPMA server, showing the secondary structural model components: alpha helix, beta turn, extended strands, and random coil. The small rectangular box shows the mutant residue
Alopecia intellectual disability syndromes 4 (APMR4) is a very rare autosomal recessive condition caused by a mutation in the LSS gene present on chromosome 21. This syndrome has a clinical heterogeneity mainly exhibited with variable degrees of intellectual disability (ID) and congenital alopecia, as well. Eight families with 13 cases have been previously reported. Herein, we provide a report on an Egyptian family with two affected siblings and one affected fetus who was diagnosed prenatally. Whole-exome sequencing (WES) revealed a novel pathogenic missense variant (c.1609G > T; p.Val537Leu) in the lanosterol synthase gene (LSS) related to the examined patients. The detected variant was confirmed by Sanger sequencing. Segregation analyses confirmed that the parents were heterozygous. Our patient was presented with typical clinical manifestations of the disease in addition to new phenotypic features which included some dysmorphic facies as frontal bossing and bilateral large ears, as well as bilateral hyperextensibility of the fingers and wrist joints, short stature, umbilical hernia, and teeth mineralization defect. This study is the first study in Egypt and the 9th molecularly proven family to date. The aim is to expand the clinical and mutational spectrum of the syndrome. Moreover, the report gives a hint on the importance of prenatal testing and the proper genetic counseling to help the parents to take their own decision based on their beliefs.
Insulin signaling disruption and caspase-3 cleavage play a pathologic role in Alzheimer’s disease (AD). Evidence suggested that cinnamaldehyde (Cin), the major component of cinnamon, has the ability to act as a neuroprotective agent. However, little evidence is available to demonstrate its effectiveness in regulating the insulin and caspase-3 signaling pathways and underlying molecular mechanisms. Therefore, the present study was conducted to correlate the molecular mechanisms of these signaling pathways and Cin treatment on animal behavioral performance in an intracerebroventricular (ICV)-streptozotocin (STZ, 3 mg/kg) model. The sporadic AD rat model was treated with Cin (10 and 100 mg/kg; intraperitoneal, i.p) daily for 2 weeks. Novel object recognition (NOR), Morris water maze (MWM), and elevated plus maze (EPM) tests were performed to assess recognition/spatial memory and anxiety-like behavior, respectively. Hippocampal Aβ aggregation was assessed using Congo red staining. The activity of hippocampal caspase-3 and IRS-1/Akt/GSK-3β signaling pathways were analyzed using the Western blot technique. The results revealed that Cin (100 mg/kg, effective dose) improved recognition/spatial memory deficits and anxiety-like behavior. In addition, Cin negated the effects of STZ on Aβ aggregation and caspase-3 cleavage in the hippocampus. Furthermore, the Western blot method showed that hippocampal IRS-1/AKT/GSK-3β phosphorylation was altered in ICV-STZ animal model, while Cin modulated this signaling pathway through decreasing Phospho.IRS-1Ser307/Total.IRS-1 ratio and also increasing Phospho.AktSer473/Total.Akt and Phospho.GSK-3βSer9/Total.GSK-3β ratios. These findings suggest that Cin is involved in the regulation of hippocampal IRS-1/AKT/GSK-3β and caspase-3 pathways in a sporadic AD model, and modulation of these signaling pathways also influences the animal behavioral performance. Graphical Abstract
Flowchart (Page et al. 2021) illustrates evaluation process of studies included in this study
Forest plots for the associations of statistically significant (p < 0.05) models with sporadic AVM risk. a allelic fixed-effect model of rs1333040; b recessive fixed-effect model of rs11169953; c recessive fixed-effect model of rs2071219
An arteriovenous malformation (AVM) is an abnormal nidus of blood vessels that is characterized by a direct connection between arteries and veins without intervening in the capillary network. The exact underlying cause of sporadic AVMs is unknown, but many studies have reported genetic associations between genes that contribute to angiogenesis, vasculogenesis, and inflammation. Eleven studies retrieved from Medline Complete, PubMed, and Google Scholar up to February 2022 were included. Heterogeneity was assessed using I² and Q-tests. Publication bias was also assessed for the shortlisted CDKN2B-AS1 rs1333040 (T > C), ACVRL1 rs2071219 (A > G), and rs11169953 (C > T) polymorphisms. The rs1333040 polymorphism showed a lower association with sporadic brain AVM for T versus C in an allelic model (OR = 0.59, 95% confidence interval [CI] = 0.41–0.84). In the recessive model, rs2071219 for AA + AG vs. GG was OR = 0.62, 95% CI = 0.43–0.9. In the recessive model, rs11169953 CC + CT vs. TT was OR = 0.56, 95% CI = 0.33–0.95. In summary, the results of this study support the association between CDKN2B-AS1 and ACVRL1 polymorphisms and sporadic brain arteriovenous malformations. This study summarized the existing information and showed the need for more replication studies on the genetic basis of sporadic AVM. In the future, more genome-wide studies should be conducted to validate and fill existing gaps in knowledge about the mechanisms of sporadic AVM development.
Cerebral small vessel disease is a common neurological disease, and its incidence is increasing year by year worldwide. In recent years, research on cerebral small vessel disease has gained more and more attention. Our research aims to visualize publications to identify the hotspots and frontiers of cerebral small vessel disease research, and to provide reference and guidance for further research. Publications related to cerebral small vessel disease were searched from the Web of Science Core Collection and screened according to inclusion criteria. CiteSpace 5.8.R3 was used to evaluate and visualize results, including generating web maps and analyzing annual publications, countries, institutions, bibliographic and co-cited references, and keywords; in this article, we use CiteSpace and VOSviewer for the 2012 Cerebral small vessel disease and bibliometric analysis from January 1, 2022 to April 30, 2022. A total of 3037 papers related to cerebral small vessel disease were retrieved, and the number of published papers showed a steady upward trend. Among them, Neuroimaging standards for research into small vessel disease and its contribution to ageing and neurodegeneration, the most symbolic references in the field of cerebral small vessel disease have been cited a total of 438 times. Stroke is the most active journal (227 articles) and USA publishes up to 800 articles. Harvard Med SchUniv Edinburgh (133 papers) and Charidimou (85 papers) are the institutions and authors who have made the most contributions in this field, respectively. Among the keywords, most of them are related to the pathogenesis of cerebral small vessel disease. After 2018, gut-brain axis and cortex are the keywords with the strongest number of cited outbreaks. There is increasing evidence that cerebral small vessel disease is a research frontier and may remain a research hotspot in the future.
Neurexin1 gene is essential for formulating synaptic cell adhesion to establish synapses. In a previous work, 38 SNPs in Neurexin1 recoded in mental disorder patients have been collected. Five computational prediction tools have been used to predict the effect of SNPs on protein function and stability. Only four SNPs in Neurexin1α have deleterious prediction results from at least four tools. The current work aims to use molecular dynamic simulation (MD) to study the effects of the four mutations on Neurexin1α both on the whole protein as well as identifying affected domains by mutations. A protein model that consists of five domains out of six domains in the real protein was used; missing residues were added, and model was tested for quality. The MD experiment has last for 1.5 μs where four parameters have been used for studying the whole protein in addition to three more parameters for the domain analysis. The whole protein study has shown that two mutations E427I for Autism and R525C for non-syndromic intellectual disability (NSID) have distinctive behavior across the four used parameters. Domain study has confirmed the previous results where the five domains of R525C have acted differently from wild type (WT), while E427I has acted differently for four domains from wild type. The other two mutations D104H and G379E have three domains that only acted differently from wild type. The fourth domain of all mutations has an obvious distinctive behavior from wild type. Further study of E427I and R525C mutations can lead to better understanding of autism and NSID.
Amyotrophic lateral sclerosis (ALS) is a fatal disease of motor neurons that mainly affects the motor cortex, brainstem, and spinal cord. Under disease conditions, microglia could possess two distinct profiles, M1 (toxic) and M2 (protective), with the M2 profile observed at disease onset. SOD1 (superoxide dismutase 1) gene mutations account for up to 20% of familial ALS cases. Comparative gene expression differences in M2-protective (early) stage SOD1G93A microglia and age-matched SOD1G93A motor neurons are poorly understood. We evaluated the differential gene expression profiles in SOD1G93A microglia and SOD1G93A motor neurons utilizing publicly available transcriptomics data and bioinformatics analyses, constructed biomolecular networks around them, and identified gene clusters as potential drug targets. Following a drug repositioning strategy, 5 small compounds (belinostat, auranofin, BRD-K78930611, AZD-8055, and COT-10b) were repositioned as potential ALS therapeutic candidates that mimic the protective state of microglia and reverse the toxic state of motor neurons. We anticipate that this study will provide new insights into the ALS pathophysiology linking the M2 state of microglia and drug repositioning.
Overexpression of Prosaposin (PSAP) lowers levels of α-synuclein. (a) Representative Western Blot for anti-GFP (Left) and anti-PSAP (Right) in EGFP-Overexpressing (EGFP-OE) cells and PSAP-Overexpressing (PSAP-OE) cells showing PSAP-GFP at 105 kDa, endogenous PSAP at 70 kDa, and control EGFP at 32 kDa. (b) Representative live-cell images of PSAP-OE cells for PSAP-GFP co-localized with the lysosome (Green: PSAP-GFP, Magenta: LysoTracker, scale bar 5 µm). (c) Representative Western Blot for anti-α-synuclein and anti-β-actin in EGFP-OE cells and PSAP-OE cells. (d) Quantification of Western Blot analysis showing levels of α-synuclein in EGFP-OE cells (n = 10) and PSAP-OE cells (n = 8). (e) Real-time PCR analysis showing mRNA levels of SNCA in EGFP-OE cells (n = 6) and PSAP-OE cells (n = 6). (f) Quantification of α-synuclein levels in cell culture media of EGFP-OE cells (n = 8) and PSAP-OE cells (n = 8) measured by enzyme-linked immunosorbent assay. (g) Representative Western blot for anti-Glucocerebrosidase (GCase) and anti-GAPDH in EGFP-OE cells and PSAP-OE cells. (h) Quantification of Western blot analysis showing levels of GCase in EGFP-OE cells (n = 8) and PSAP-OE cells (n = 8). (i) Non-specific GBA activity and specific GCase activity analysis in EGFP-OE cells (n = 8) and PSAP-OE cells (n = 8) treated with or without AMP-deoxynojirimycin. Data are expressed as mean ± SEM. n.s: non-significant, p > 0.05; *p < 0.05; **p < 0.01; ***p < 0.001, Mann–Whitney U test (d, e, f, h,) and Two-way ANOVA with Tukey’s multiple comparison test (i)
The decrease in levels of α-synuclein is GCase activity independent. (a) Non-specific GBA activity assay in PSAP-OE cells with or without CBE treatment (n = 8). (b) Quantification of Western blot analysis showing levels of GCase in PSAP-OE cells with or without CBE treatment (n = 8). (c) Quantification of Western Blot analysis showing levels of α-synuclein in PSAP-OE cells with or without CBE treatment (n = 8). (d) Representative Western blot for anti-GCase, anti-α-synuclein and anti-GAPDH in PSAP-OE cells with or without CBE treatment. Data are expressed as mean ± SEM. n.s: non-significant, ***p < 0.001, Mann–Whitney U test (a–c)
Overexpression of PSAP affects autophagy. (a) Representative Western blot for anti-LC3b and anti-β-actin in 100 nM Bafilomycin A1 or dimethyl sulfoxide (DMSO) treated EGFP-OE or PSAP-OE cells. (b) Quantification of Western blot analysis showing levels of LC3-II in 100 nM Bafilomycin A1 or DMSO treated EGFP-OE or PSAP-OE cells (n = 5). (c) Quantification of Western blot analysis showing LC3-II/LC3-I ratio in 100 nM Bafilomycin A1 or DMSO treated EGFP-OE or PSAP-OE cells (n = 5). (d) Representative Western blot for anti-α-synuclein and anti-β-actin in 100 nM Bafilomycin A1 or DMSO treated EGFP-OE or PSAP-OE cells. (e) Quantification of Western blot analysis showing levels of α-synuclein in 100 nM Bafilomycin A1 or DMSO treated EGFP-OE cells (n = 4). (f) Quantification of Western blot analysis showing levels of α-synuclein in 100 nM Bafilomycin A1 or DMSO treated PSAP-OE cells (n = 4). (g) Representative Western blot for anti-α-synuclein and anti-GAPDH in 10 µM MG132 or DMSO treated EGFP-OE or PSAP-OE cells. (h) Quantification of Western blot analysis showing levels of α-synuclein in 10 µM MG132 or DMSO treated EGFP-OE cells (n = 4). (i) Quantification of Western blot analysis showing levels of α-synuclein in 10 µM MG132 or DMSO treated PSAP-OE cells (n = 4). Data are expressed as mean ± SEM. *p < 0.05; **p < 0.01, Two-way ANOVA with Tukey’s multiple comparison test (b, c) and unpaired t-test (e, f, h, i)
siRNA knockdown of PSAP increases levels of α-synuclein. (a) Representative Western blot for anti-PSAP and anti-β-actin in 10 nM siCTL or siPSAP treated WT SH-SY5Y cells. Endogenous PSAP is detected as an intracellular form of 68 kDa and deglycosylated form of ~ 50 kDa (Kishimoto et al. 1992). (b) Quantification of Western blot analysis showing levels of PSAP in siCTL or siPSAP treated WT SH-SY5Y cells (n = 8). (c) Real-time PCR analysis showing mRNA levels of PSAP in siCTL or siPSAP treated WT SH-SY5Y cells (n = 8). (d) Representative Western blot for anti-α-synuclein and anti-β-actin in siCTL or siPSAP treated WT SH-SY5Y cells. (e) Quantification of Western blot analysis showing levels of α-synuclein in siCTL or siPSAP treated WT SH-SY5Y cells (n = 8). (f) Real-time PCR analysis showing mRNA levels of SNCA in siCTL or siPSAP treated WT SH-SY5Y cells (n = 8). Data are expressed as mean ± SEM. n.s: non-significant, p > 0.05; *p < 0.05; **p < 0.01; ***p < 0.001, Mann–Whitney U test (b, c, e, f)
Saposin C can detach α-synuclein from glucosylceramide-enriched lipid vesicles. (a, b) Representative Western blot for anti-α-synuclein in following treatment of the vesicles; incubated with recombinant α-synuclein and control peptide (Control) or short synthetic saposin C peptide (TX14(a)) or recombinant saposin C (Saposin C) in either pH 5.4 or 7.4 buffer. After incubation, vesicles were separated into retained and flow-through fractions by centrifugation. Left blots show α-synuclein retained with vesicles and right blots show dislodged or unbound α-synuclein detected in flow-through. (c–e) Quantification of Western blot analysis from 3 independent experiments showing the amount of α-synuclein in retained fraction (c), flow-through fraction (d) and the ratio of flow-through over-retained α-synuclein (e) at lysosomal pH (pH 5.4). (f–h) Quantification of Western blot analysis from 3 independent experiments showing the amount of α-synuclein in retained fraction (f), flow-through fraction (g) and the ratio of flow-through over-retained α-synuclein (h) at cytoplasmic pH (pH 7.4). Data are expressed as mean ± SEM. n.s: non-significant, p > 0.05; *p < 0.05, n = 3 in each group, One-way ANOVA with Dunnett's multiple comparison test (c–h)
Parkinson’s disease (PD) is the second most common progressive neurodegenerative disorder affecting over 1% of the 65 + age population. Saposin C, a lysosomal protein required for the normal activity of glucocerebrosidase (GCase), may serve as a disease modifier in PD. Saposin C is cleaved from its precursor, Prosaposin (PSAP), which is secreted as an uncleaved protein and exerts neuroprotective effects. In this study, we aim to elucidate the neuroprotective roles of PSAP and saposin C in PD by evaluating their effects on α-synuclein accumulation in human neuroblastoma cells. Stable overexpression of PSAP reduced monomeric α-synuclein levels in SH-SY5Y cells, while PSAP knockdown by small interfering RNA led to the opposite effect, and those effects were independent of GCase activity. Autophagy flux was decreased by stable PSAP overexpression. Furthermore, a flow-through assay revealed that recombinant saposin C was able to detach α-synuclein from artificial glucosylceramide-enriched lipid membranes at the lysosomal pH. Taken together, our findings provide further evidence that PSAP and saposin C as key proteins involved in α-synuclein clearance by dislodging it from lipid membranes.
Oxidative stress is considered a possible mechanism in Parkinson’s disease (PD) progression. Bilirubin has been recognized as a powerful antioxidant that increases due to heme-oxygenase activity. We aimed to investigate the association of total bilirubin (TB) with motor signs and asymmetry in different stages of early PD. A case–control study was performed to investigate the differences in TB levels in PD patients and healthy controls (HC) both carrying LRRK2 variants. We compared TB levels in HC and Hoehn and Yahr (HY) I and II cohorts separately, followed by multiple linear regression analysis to evaluate the association between TB and motor dysfunction in each stage. We used Movement Disorder Society-Sponsored Revision of the Unified Parkinson’s Disease Rating Scale (UPDRS) part III scores and asymmetry scores to address motor disability. Asymmetry scores were calculated from the corresponding UPDRS III tasks. TB was significantly increased in HY II compared to HC (P < 0.001). Positive correlations with TB were found for UPDRS III total score (ρ = 0.303, P = 0.034) and asymmetry score (ρ = 0.418, P = 0.003) in HY I. Multiple linear regression found a significant relationship between TB and asymmetry scores in HY I (R2 = 0.261, P = 0.037), but no relationship was achieved with UPDRS III total scores. Increased TB serves as an important diagnostic marker in earlier stages of PD. A significant relationship was found between TB and motor asymmetry in HY I patients. According to our findings, bilirubin mainly exhibits its protective effects in HY I population.
Ischemic stroke (IS) poses a heavy burden on the healthcare system, and revascularization is the most effective treatment. However, ischemia/reperfusion (I/R) injury, one main cause of revascularization complications, significantly hinders IS recovery. Unfortunately, none of the neuroprotectants tested to date has been successfully translated clinically for post-revascularization I/R injury therapy. In multiple pathophysiological processes, apoptosis antagonizing transcription factor (AATF) serves as a cell protector, but its role in neuronal I/R injury is unknown. Therefore, we firstly demonstrated the expression profiles of AATF in a distal middle cerebral artery occlusion/reperfusion (dMCAO/R) model and found that AATF expression was increased in cortical neuron after dMCAO/R. Over-expressing AATF reduced infarct volume, alleviated neuronal death, and promoted neurological functions. Next, we used an oxygen–glucose deprivation/reoxygenation (OGD/R) model to investigate the mechanism of AATF. Results indicated that AATF alleviated OGD/R-induced large-scale DNA fragmentation, which suggested that the protective effect of AATF may be attributed to parthanatos inhibition. After that, we examined the regulatory mechanism of AATF. We found that AATF did not affect poly (ADP-ribose) accumulation and apoptosis-inducing factor (AIF) nucleus translocation. AATF competitively interacted with nuclear AIF, which inhibited AIF from binding DNA. At last, we verified the effect and mechanism of AATF in dMCAO/R model. The present study, for the first time, demonstrates the expression, function, and mechanism of AATF in the context of neuronal I/R injury via dMCAO/R and OGD/R model, which provides new evidence in this area and may facilitate exploring new therapeutic targets.
The effect of prenatal METH (2 and 5 mg/kg) exposure on expression of the Bax, Bcl-2, procaspase-3, and cleaved caspase-3 in the prefrontal cortex (A, B, and C) and amygdala (D, E, and F) of adult rats’ brain. Values are presented as mean ± SEM. (N = 4; *, #, and ^ different from the control, METH 2 mg/kg and sex; *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001)
The effect of prenatal METH (2 and 5 mg/kg) exposure on expression of the Beclin-1, LC3I, and LC3I in the prefrontal cortex (A, B, and C) and amygdala (D, E, and F) of adult rats’ brain. Values are presented as mean ± SEM. (N = 4; *, #, and ^ different from the control, METH 2 mg/kg and sex; *P < 0.05, ***P < 0.001, and ****P < 0.0001)
Experimental procedure of study and molecular pathways leading to apoptosis and autophagy in the brain of prenatal METH (2 and 5 mg/kg) exposed offspring in adulthood
In the last decade, there has been a great increase in methamphetamine hydrochloride (METH) abuse by pregnant women that exposes fetus and human offspring to a wide variety of developmental impairments that may be the underlying causes of future psychosocial issues. Herein, we investigated whether prenatal METH exposure with different doses (2 and 5 mg/kg) could influence neuronal cell death and antioxidant level in the different brain regions of adult male and female offspring. Adult male and female Wistar rats prenatally exposed to METH (2 or 5 mg/kg) and/or saline was used in this study. At week 12, adult rats’ offspring were decapitated to collect different brain region tissues including amygdala (AMY) and prefrontal cortices (PFC). Western blot analysis was performed to evaluate the apoptosis- and autophagy-related markers, and enzymatic assay was used to measure the level of catalase and also reduced glutathione (GSH). Our results showed that METH exposure during pregnancy increased the level of apoptosis (BAX/Bcl-2 and Caspase-3) and autophagy (Beclin-1 and LC3II/LC3I) in the PFC and AMY areas of both male and female offspring’s brain. Also, we found an elevation in the GSH content of all both mentioned brain areas and catalase activity of PFC in the offspring’s brain. These changes were more significant in female offspring. Being prenatally exposed to METH increased cell death at least partly via apoptosis and autophagy in AMY and PFC of male and female offspring’s brain, while the antioxidant system tried to protect cells in these regions. Graphical Abstract
Intracerebral hemorrhage causes high mortality and morbidity, but its therapy methods are limited. In the present study, pulsed electromagnetic field (PEMF) was demonstrated to have beneficial effects on an intracerebral hemorrhage (ICH) model. This study explored the effects and underlying mechanisms of PEMF in a mouse model of ICH and cultured BV2 cells. PEMF was applied 4 hours after collagenase-induced ICH at day 0 and 4 hours per day for seven consecutive days. The expression levels of proinflammatory factors were assessed by ELISA kits and western blotting. Hematoma volume was measured by histological analysis. The effects of PEMF on phagocytosis of the erythrocytes were observed in cultured BV2 cells and ICH mouse models. Seven days after ICH, the hematoma volume was significantly reduced in PEMF-treated animals compared to nontreated mice. We found that PEMF decreased the hematoma volume and the expression levels of proinflammatory factors after ICH. Moreover, PEMF enhanced the erythrophagocytosis of microglia via CD36. Furthermore, we found that downregulation CD36 with Genistein blocked the effects of PEMF-induced hematoma clearance and anti-inflammations effects. Thus, the PEMF-mediated promotion of neurological functions may at least partly involve anti-inflammatory processes and hematoma clearance. These results suggest that PEMF treatment promoted the hematoma clearance and alleviated the inflammation after ICH.
Given the complexity of pathophysiological processes of brain tumors, ineffective therapies, and high mortality rate, new therapeutic options with less toxicity are necessary. Hyssopus officinalis (hyssop) is an aromatic plant with important biological activities. The aim of this study is to assess the anti-cancer effect of hyssop extract on damages of glioblastoma multiforme. In this study, total flavonoids, phenolic content, and quantification of phenolic compound of hyssop extracts were analyzed. In vitro antioxidant properties of hyssop extract were also examined. In addition, cell viability, apoptosis, and cell cycle were evaluated in C6 glioma cell culture. In vivo, the rats were divided randomly into four main groups: intact, control, vehicle, and treatment groups. 1 × 10⁶ C6 rat glioma cells were implanted into the right caudate nucleus of the rat’s brain. The treatment group received the methanol extract of hyssop (100 mg/kg) for 7 days. Evolution of locomotor activity, tumor volume, survival rate, activities of antioxidant enzymes (superoxide dismutase (SOD) and catalase (CAT)), vascular endothelial growth factor (VEGF) expression, TUNEL-positive cells, p53 and p21 mRNA expression, and histological alterations were performed. The results showed that the methanol extract of hyssop increased the apoptosis and reduced the cell division of C6 glioma cells in cell culture. Moreover, methanol extract decreased the tumor volume and prolonged survival. Also, the activity of SOD and CAT enzymes was reduced in tumor tissue and enhanced in surrounding tissue. TUNEL-positive cells were increased in methanol extract of hyssop group. The expression of p53 and p21 mRNA was upregulated in the treatment group. Moreover, the histological analysis indicated a considerable decrease in invasion of tumor cells and inflammation in the hyssop-treated rats. According to the achieved results, it can be stated that hyssop has sufficient potential to inhibit damage of brain tumors, at least in part, by affecting the oxidative stress and cell proliferation pathways.
SET and MYND domain-containing protein 2 (SMYD2) is an important epigenetic regulator that methylates histone and non-histone proteins. The study aimed to investigate the oncogenic role of SMYD2 in gliomas and explore its degradation mechanism induced by cisplatin. Tumor tissue microarray of 441 patients with glioma was collected for SMYD2 immunohistochemical staining. Kaplan–Meier survival curves were constructed using the overall survival values. mRNA-sequencing analysis was performed for understanding the downstream mechanisms mediated by SMYD2. The half-inhibitory concentrations (IC50) of temozolomide and cisplatin in AZ505-treated and control cells were calculated. The potential E3 ubiquitin ligase of SMYD2 was predicted in UbiBrowser and confirmed by a knockdown test. The effect of SMYD2 and its E3 ligase on apoptosis and migration of glioma cells was determined via cell-function assays. High SMYD2 expression correlated with a high WHO stage (P = 0.004) and a low survival probability (P = 0.012). The inhibition of SMYD2 suppressed the process of epithelial to mesenchymal transition (EMT) by downregulating the expression of Collagen 1A1 (COL1A1). AZ505 treatment significantly increased the drug sensitivity of glioma cells. SMYD2 expression was markedly reduced by cisplatin treatment via STIP1 Homology And U-Box Containing Protein 1 (STUB1)-mediated degradation. The knockdown of STUB1 could partly reverse the cell function impairment induced by cisplatin. Our findings suggested that SMYD2 could be a potential drug target for the treatment of gliomas, and STUB1-mediated degradation of SMYD2 plays an important role in reversing chemotherapy resistance in patients with gliomas.
Workflow of this study
HE staining assessment of neuronal morphology in the hippocampal CA1 area. A The sham group showed an orderly arrangement of neurons with complete cell structure; B the I/R 12 h group showed a reduced number of neurons, but their structure was complete, and there were no typical apoptotic cells; C the I/R 48 h group showed a disorderly arrangement of cells, and the cytoplasm and nucleoli were stained deeply, indicating nuclear pyknosis. Scale bars = 50 µm
Functional analysis of differentially expressed proteins at 12 h and 48 h after I/R. A Biological process category; B molecular function category; C cellular component category
STRING PPI network analysis of the 164 differentially expressed proteins in I/R rats. The average node degree was 3.73, the average local clustering coefficient was 0.466, and the PPI enrichment p-value was less than 1.0E − 16
Abundance of candidate urine biomarkers in I/R rats by PRM quantification. The x‐axis represents different groups, and the y‐axis represents the area of intensity based on PRM quantification. * p < 0.05
Cerebral ischemia–reperfusion (I/R) injury is the leading cause of death in severe hypotension caused by cardiac arrest, drowning, and excessive blood loss. Urine can sensitively reflect pathophysiological changes in the brain even at an early stage. In this study, a rat model of global cerebral I/R injury was established via Pulsinelli’s four-vessel occlusion (4-VO) method. Overall, 164 urinary proteins significantly changed in the 4-VO rat urine samples compared to the control samples by data-independent acquisition (DIA) proteomics technique (1.5-fold change, p < 0.05). Gene Ontology annotation showed that the acute-phase response, the ERK1 and ERK2 cascade, endopeptidase activity, blood coagulation, and angiogenesis were overrepresented. After parallel reaction monitoring (PRM) validation, 15 differential proteins having human orthologs were verified as the potential urinary markers associated with cerebral I/R injury. Of these potential biomarkers, 8 proteins were reported to be closely associated with cerebral I/R injury. Nine differential proteins changed even when there were no clinical manifestations or histopathological cerebral damage, including FGG, COMP, TFF2, HG2A, KNG1, CATZ, PTGDS, PRVA, and HEPC. These 9 proteins are potential biomarkers for early screening of cerebral I/R injury to prevent the development of cerebral injury. KNG1, CATZ, PTGDS, PRVA, and HEPC showed an overall trend of upregulation or downregulation at 12 and 48 h after I/R injury, reflecting the progression of cerebral I/R injury. These 5 proteins may serve as potential biomarkers for prognostic evaluation of cerebral I/R injury. These findings provide important clues to inform the monitoring of cerebral I/R injury and further the current understanding of its molecular biological mechanisms.
Kaplan–Meier (KM) overall survival (OS) analysis representing case IDs with BCR recombination read recoveries from primary tumor RNA-seq files. A Data represent the single value physicochemical parameter, omega, for IGK; median survival of top 50th percentile (n = 71) (grey line) of omega values, 51 months; median survival of bottom 50th percentile (n = 71) (black line) of omega values, 80 months. Log-rank p value, 0.0463. B Data represent the single value physicochemical parameter, instability index, for IGK; median survival of top 50th percentile (n = 71) (grey line) of instability index values, 125 months; median survival of bottom percentile (n = 71) (black line) of instability index values, 51 months. Log-rank p value, 0.0418. C Data represent the single value physicochemical parameter, omega, for IGL; median survival of top 50th percentile (n = 73) (grey line) of omega values, 51 months; median survival of bottom 50th percentile (n = 73) (black line) of omega values, 125 months. Log-rank p value, 0.0102. D Data represent the single value physicochemical parameter, PPII propensity, for IGH; median survival of top 50th percentile (n = 74) (grey line) of PPII values, 37 months; median survival of bottom 50th percentile (n = 73) (black line) of PPII values, 125 months. Log-rank p value, 0.0008
KM OS analysis representing case IDs with BCR light chain CDR3 AA chemical sequence motifs from primary tumor RNA-seq files. Case IDs representing IGK CDR3s containing the AA chemical sequence motifs (A), NXHXHA (black line) and (B), ANXHXHA (black line) (n = 28) represent a higher OS when compared to AR (grey line) (n = 114); log-rank p value, 0.0258. Note, that the (A) motif is contained within the (B) and the two motifs represent identical case IDs. Case IDs representing IGL CDR3s containing the AA chemical sequence motifs (C), XAHXXAA (black line) (n = 27), compared to AR (grey line) (n = 119), and (D), XNXXXHN (black line) (n = 20), compared to AR (grey line) (n = 126). In both cases, the indicated chemical sequence motif represents a higher OS when compared to AR (grey); log-rank p values, 0.0209, 0.0150, respectively
KM OS analysis representing case IDs representing combo homology groups (Methods) for the BCR CDR3s. A For the first IGH CDR3 homology analysis, log-rank p value, 0.0380; median survival for out-group-1 (n = 69, grey line), 79 months; in-group-1 (n = 17, black line), 34 months. B For the second IGH CDR3 homology analysis, log-rank p value, 0.0415; median survival for out-group-2 (n = 32, grey line), undefined; median survival for in-group-2 (n = 54, black line), 46 months. C For the first IGK CDR3 homology analysis log-rank p value, 0.0466; median survival for out-group-1 (n = 54, grey line), 46 months; median survival for in-group-1 (n = 19, black line), 125 months. D For the second IGK CDR3 homology analysis log-rank p value, 0.0401; median survival for out-group-2 (n = 57, grey line), 46 months; median survival for in-group-2 (n = 16, black line), 125 months. E For the IGL CDR3 homology analysis, log-rank p value, log-rank p value, 0.0361; median survival for out-group (n = 54, grey line), undefined; median survival for in-group (n = 46, black line), 46 months
Pediatric neuroblastoma (NBL) is one of the most common pediatric cancers, and it can often be aggressive. Genetic and demographic factors can correlate with the severity of NBL, but the variations in the B-cell receptors (BCRs) or immunoglobulin proteins present in the NBL tumors, and their relationships to survival, are not well understood. BCRs contain variations in their complementary determining region-3 (CDR3s) amino acid sequences, due to variable recombinations of the V- and J-gene segments. Accordingly, these variations in CDR3s may represent different antigen interactions and thereby different survival probabilities. Thus, we mined the TARGET project, NBL tumor RNAseq files for BCR recombination reads. Evaluations of the physicochemical properties of IGK, IGL, and IGH CDR3s from these tumors pointed to properties of IGK and IGL in particular as associated with survival distinctions, based on several independent bioinformatics approaches, including a novel homology grouping approach facilitated by a recently developed web tool, In conclusion, tumor resident BCR chemical features are likely useful for better risk stratification and for guiding therapy, and the availability of a user-friendly web tool will likely facilitate using BCR chemical features to meet those goals.
PNU-282987, a selective alpha7 nicotinic acetylcholine receptor agonist, has previously been shown to have both neurogenic and broad regenerative effects in the adult murine retina. The objective of this study was to assay the molecular mechanism by which PNU-282987 promotes the production of Muller-derived progenitor cells through signaling via the resident retinal pigment epithelium. These Muller-derived progenitor cells generate a myriad of differentiated neurons throughout the retina that have previously been characterized by morphology. Herein, we demonstrate that topical application of PNU-282987 stimulates production of functional neurons as measured by electroretinograms. Further, we examine the mechanism of how this phenomenon occurs through activation of this atypical receptor using a transcriptomic approach isolated retinal pigment epithelium activated by PNU-282987 and in whole retina. We provide evidence that PNU-282987 causes a bi-modal signaling event in which early activation primes the retina with an inflammatory response and developmental signaling cues, followed by an inhibition of gliotic mechanisms and a decrease in the immune response, ending with upregulation of genes associated with specific retinal neuron generation. Taken together, these data provide evidence that PNU-282987 activates the retinal pigment epithelium to signal to Muller glia to produce Muller-derived progenitor cells, which can differentiate into new, functional neurons in adult mice. These data not only increase our understanding of how adult mammalian retinal regeneration can occur, but also provide therapeutic promise for treating functional vision loss.
Mild cognitive impairment (MCI) is a pathological stage between normal cognitive aging and dementia. The blood–brain barrier (BBB) breakdown is emerging as an early biomarker of MCI. This study aimed to visually analyze the literatures related to MCI caused by BBB dysfunction in recent 20 years, systematically identify collaboration networks, track research trends, highlight current hot spots, and predict future frontiers in this field. The related literatures published from 2000 to 2021 were obtained from the Web of Science with the search term “mild cognitive impairment and (blood–brain barrier or neurovascular unit or neurovascular coupling)”. VOSviewer software was used to present knowledge map, CiteSpace software was used to extract literature information and make tables, including the top most influential countries, authors, institutions, periodicals, keywords, and references. A total of 333 literatures were used for visual analysis. After 2013, the literatures in the field of BBB dysfunction-induced MCI showed an increased trend in terms of year of publication and quantity of material, with more than 40 publications published each year. USA, England, China, and Sweden cooperated closely. In terms of institutions, Harvard Med Sch ranked first in the number of papers published, followed by Mstricht Univ and Univ Washington. In terms of journals, three of the top five co-cited journals belonged to USA, the other two journals were Neurobiol Aging and J Alzheimers DIS, which were from England and Netherlands respectively. A total of 1752 authors were identified, with Abhay P Sagare the most published and Zlokovic BV the most cited. Keyword emergence detection analysis showed that inflammation, oxidative stress, and memory were new research hot spot in this field. Overall, the research on BBB dysfunction-induced MCI is booming. In the future, cooperation and communication between different countries and institutions should be strengthened.
Altered amine transporter function, phosphorylation, and association with interacting proteins are evident in animals with a history of psychostimulant exposure. Our previous studies have shown that the Thr258/Ser259 motif in the norepinephrine transporter (NET) is involved in amphetamine (AMPH)-mediated NET regulation and behavior. However, the neurobiological consequences of in vivo Thr258/Ser259-dependent NET regulation in an intact animal model are unclear. Therefore, we generated a viable construct-valid NET-Thr258Ala/Ser259Ala (NET-T258A/S259A) mouse model using CRISPR/Cas9 technology by replacing Thr258/Ser259 motif with Ala258/Ala259 motif. NET-T258A/S259A mice have a birth rate consistent with Mendelian inheritance ratios. Both male and female homozygous NET-T258A/S259A mice are viable, display normal growth and general health, and exhibit normal body weight (sex-dependent) and total activity in the open field similar to their wild-type (WT) littermates. NET-T258A/S259A mice showed reduced NET function in the prefrontal cortex (PFC) compared to WT mice while NET function in the nucleus accumbens (NAc) remained unchanged. Compared to respective WT counterparts, NET-T258A/S259A males but not females showed significantly reduced locomotor activation in response to acute AMPH administration and significantly reduced AMPH-induced conditioned place preference (CPP). When tested in the males only, acute AMPH administration inhibited NET function and surface expression in the WT NAc but not in the NET-T258A/S259A NAc while AMPH administration inhibited DAT function and surface expression in the NAc of both WT and NET-T258A/S259A mice. Collectively, our findings reveal that the mice carrying the T258A/S259A mutation in NET gene display brain region-specific differences in NET functional expression and blunted response to AMPH.
Schematic representation of chemokines and their receptors
Changes in expression of chemokines and chemokine receptors in humans
Effects of chemokine and chemokine receptor manipulations in rodent models of AD
Alzheimer’s disease (AD) is a progressive neurogenerative disorder manifested by gradual memory loss and cognitive decline due to profound damage of cholinergic neurons. The neuropathological hallmarks of AD are intracellular deposits of neurofibrillary tangles (NFTs) and extracellular aggregates of amyloid β (Aβ). Mounting evidence indicates that intensified neuroinflammatory processes play a pivotal role in the pathogenesis of AD. Chemokines serve as signaling molecules in immune cells but also in nerve cells. Under normal conditions, neuroinflammation plays a neuroprotective role against various harmful factors. However, overexpression of chemokines initiates disruption of the integrity of the blood–brain barrier, facilitating immune cells infiltration into the brain. Then activated adjacent glial cells–astrocytes and microglia, release massive amounts of chemokines. Prolonged inflammation loses its protective role and drives an increase in Aβ production and aggregation, impairment of its clearance, or enhancement of tau hyperphosphorylation, contributing to neuronal loss and exacerbation of AD. Moreover, chemokines can be further released in response to growing deposits of toxic forms of Aβ. On the other hand, chemokines seem to exert multidimensional effects on brain functioning, including regulation of neurogenesis and synaptic plasticity in regions responsible for memory and cognitive abilities. Therefore, underexpression or complete genetic ablation of some chemokines can worsen the course of AD. This review covers the current state of knowledge on the role of particular chemokines and their receptors in the development and progression of AD. Special emphasis is given to their impact on forming Aβ and NFTs in humans and in transgenic murine models of AD.
Multiple sclerosis (MS) is a common chronic autoimmune disorder of the central nervous system that predominantly affects young adults. Mounting evidence indicates that deregulation of microRNAs (miRNAs) in cerebrospinal fluid (CSF) has been implicated in MS as a potential biomarker. However, comprehensive assessments of CSF miRNAs and their target genes are lacking. Here, aberrantly expressed CSF miRNAs of MS patients were obtained from numerous studies by manual search. With detailed information on these miRNAs, we utilized online databases to screen out immune-related target genes and further performed Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. To identify MS high-risk pathways and pivotal genes, pathway crosstalk and pathway-gene networks were constructed, followed by the establishment of a protein-protein interaction (PPI) network. The datasets collected from ArrayExpress were used to assess pivotal genes. Overall, 21 MS-related CSF miRNAs were included in this study. Subsequently, we identified 469 MS-related genes and 14 high-risk pathways. In the pathway-gene network, 27 critical MS-related genes participated in at least half of the high-risk pathways, and these genes were used to identify pivotal genes. Finally, miR-150, miR-328, and miR-34c-5p were determined to be risk miRNAs via the regulation of the pivotal risk genes MAPK1, AKT1, and VEGFA. Among them, VEGFA was validated to be significantly decreased in the CSF cells of MS patients by transcriptomic datasets. These findings may provide potential biomarkers or therapeutic targets and help elucidate the molecular mechanisms underlying the pathogenesis of MS.
Ketamine is a noncompetitive antagonist of N-methyl-D-aspartate (NMDA) receptors. Many experimental studies have shown that ketamine can induce cognitive impairments and schizophrenia-like symptoms. While much data have demonstrated that glial cells are associated with the pathophysiology of psychiatric disorders, including schizophrenia, the response of glial cells to ketamine and its significance to schizophrenia are not clear. The present study was intended to explore whether chronic ketamine treatment would induce behavioral and glial changes in mice. First, ketamine was used to stimulate behavioral abnormalities similar to schizophrenia evaluated by the open field test, elevated plus-maze test, Y maze test, novel object recognition test, and tail suspension test. Secondly, histopathology and Nissl staining were performed. Meanwhile, immunofluorescence was used to evaluate the expression levels of IBA-1 (a microglial marker) and GFAP (an astrocyte marker) in the mouse hippocampus for any change. Then, ELISA was used to analyze proinflammatory cytokine levels for any change. Our results showed that ketamine (25 mg/kg, i.p., qid, 12 days) induced anxiety, recognition deficits, and neuronal injury in the hippocampus. Moreover, chronic ketamine treatment enhanced GFAP expression in CA1 and DG regions of the hippocampus but did not influence the expression of IBA-1. Ketamine also increased the levels of IL-1β, IL-6, and TNF-α in the mouse hippocampus. Our study created a new procedure for ketamine administration, which successfully induce negative symptoms and cognitive-behavioral defects in schizophrenia by chronic ketamine. This study further revealed that an increase in astrocytosis, but not microglia, is associated with the mouse model of schizophrenia caused by ketamine. In summary, hippocampal astrocytes may be involved in the pathophysiology of ketamine-induced schizophrenia-like phenotypes through reactive transformation and regulation of neuroinflammation.
Postoperative cognitive dysfunction (POCD) is a cognitive deterioration and dementia that arise after a surgical procedure, affecting up to 40% of surgery patients over the age of 60. The precise etiology and molecular mechanisms underlying POCD remain uncovered. These reasons led us to employ integrative bioinformatics and machine learning methodologies to identify several biological signaling pathways involved and molecular signatures to better understand the pathophysiology of POCD. A total of 223 differentially expressed genes (DEGs) comprising 156 upregulated and 67 downregulated genes were identified from the circRNA microarray dataset by comparing POCD and non-POCD samples. Gene ontology (GO) analyses of DEGs were significantly involved in neurogenesis, autophagy regulation, translation in the postsynapse, modulating synaptic transmission, regulation of the cellular catabolic process, macromolecule modification, and chromatin remodeling. Pathway enrichment analysis indicated some key molecular pathways, including mTOR signaling pathway, AKT phosphorylation of cytosolic targets, MAPK and NF-κB signaling pathway, PI3K/AKT signaling pathway, nitric oxide signaling pathway, chaperones that modulate interferon signaling pathway, apoptosis signaling pathway, VEGF signaling pathway, cellular senescence, RANKL/RARK signaling pathway, and AGE/RAGE pathway. Furthermore, seven hub genes were identified from the PPI network and also determined transcription factors and protein kinases. Finally, we identified a new predictive drug for the treatment of SCZ using the LINCS L1000, GCP, and P100 databases. Together, our results bring a new era of the pathogenesis of a deeper understanding of POCD, identified novel therapeutic targets, and predicted drug inhibitors in POCD.
KLS-13019, a novel devised cannabinoid-like compound, was explored for anti-inflammatory actions in dorsal root ganglion cultures relevant to chemotherapy-induced peripheral neuropathy (CIPN). Time course studies with 3 µM paclitaxel indicated > 1.9-fold increases in immunoreactive (IR) area for cell body GPR55 after 30 min as determined by high content imaging. To test for reversibility of paclitaxel-induced increases in GPR55, cultures were treated for 8 h with paclitaxel alone and then a dose response to KLS-13019 added for another 16 h. This “reversal” paradigm indicated established increases in cell body GPR55 IR areas were decreased back to control levels. Because GPR55 had previously reported inflammatory actions, IL-1β and NLRP3 (inflammasome-3 marker) were also measured in the “reversal” paradigm. Significant increases in all inflammatory markers were produced after 8 h of paclitaxel treatment alone that were reversed to control levels with KLS-13019 treatment. Accompanying studies using alamar blue indicated that decreased cellular viability produced by paclitaxel treatment was reverted back to control levels by KLS-13019. Similar studies conducted with lysophosphatidylinositol (GPR55 agonist) in DRG or hippocampal cultures demonstrated significant increases in neuritic GPR55, NLRP3 and IL-1β areas that were reversed to control levels with KLS-13019 treatment. Studies with a human GPR55-β-arrestin assay in Discover X cells indicated that KLS-13019 was an antagonist without agonist activity. These studies indicated that KLS-13019 has anti-inflammatory properties mediated through GPR55 antagonist actions. Together with previous studies, KLS-13019 is a potent neuroprotective, anti-inflammatory cannabinoid with therapeutic potential for high efficacy treatment of neuropathic pain.
Ceramide accumulation has been associated with ischemic stroke. Myriocin is an effective serine palmitoyltransferase (SPT) inhibitor that reduces ceramide levels by inhibiting the de novo synthesis pathway. However, the role of myriocin in cerebral ischemia/reperfusion (I/R) injury and its underlying mechanism remain unknown. The present study established an experimental rat model of middle cerebral artery occlusion (MCAO). We employed ultra-performance liquid chromatograph quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS)–based lipidomic analysis to identify the disordered lipid metabolites and the effects of myriocin in cerebral cortical tissues of rats. In this study, we found 15 characterized lipid metabolites involved in sphingolipid and glycerophospholipid metabolism in cerebral I/R-injured rats, and these alterations were significantly alleviated by myriocin. Specifically, the mRNA expression of metabolism-related enzyme genes was detected by real-time quantitative polymerase chain reaction (RT-qPCR). We demonstrated that myriocin could regulate the mRNA expression of ASMase, NSMase, SGMS1, SGMS2, ASAH1, ACER2, and ACER3, which are involved in sphingolipid metabolism and PLA2, which is involved in glycerophospholipid metabolism. Moreover, TUNEL and Western blot assays showed that myriocin plays a key role in regulating neuronal cell apoptosis. In summary, the present work provides a new perspective for the systematic study of metabolic changes in ischemic stroke and the therapeutic applications of myriocin.
Paralytic ileus is common in patients with septic shock, causing high morbidity and mortality. Enteric neurons and enteric glial cells (EGCs) regulate intestinal motility. However, little is known about their interaction in endotoxemia. This study aimed to investigate whether reactive EGCs had harmful effects on enteric neurons and participated in intestinal motility disorder in mice during endotoxemia. Endotoxemia was induced by the intraperitoneal injection of lipopolysaccharide (LPS) in mice. Fluorocitrate (FC) was administered before LPS injection to inhibit the reactive EGCs. The effects of reactive EGCs on intestinal motility were analyzed by motility assays in vivo and colonic migrating motor complexes ex vivo. The number of enteric neurons was evaluated by immunofluorescent staining of HuCD, nNOS, and ChAT in vivo. In addition, we stimulated EGCs with IL-1β and TNF-α in vitro and cultured the primary enteric neurons in the conditioned medium, detecting the apoptosis and morphology of neurons through staining TUNEL, cleaved caspase-3 protein, and anti-β-III tubulin. Intestinal motility and peristaltic reflex were improved by inhibiting reactive EGCs in vivo. The density of the neuronal population in the colonic myenteric plexus increased significantly, while the reactive EGCs were inhibited, especially the nitrergic neurons. In vitro, the enteric neurons cultured in the conditioned medium of reactive EGCs had a considerably higher apoptotic rate, less dendritic complexity, and fewer primary neurites. Reactive enteric glial cells probably participated in paralytic ileus by damaging enteric neurons during endotoxemia. They might provide a novel therapeutic strategy for intestinal motility disorders during endotoxemia or sepsis.
BDNF levels in COVID-19 patients and healthy individuals. A significant difference was observed in serum BDNF levels between patients and healthy individuals. *p < 0.05
BDNF levels in patients with neurological manifestations. A A significant difference was observed in serum BDNF levels between patients with neurological manifestations and healthy individuals. B BDNF serum levels in patients with and without CNS or PNS manifestations. A significant difference was observed in serum BDNF levels between patients with and without CNS manifestations. ns not significant, *p < 0.05
BDNF serum levels in patients with different requirement of supplemental oxygen. A significant difference was observed in serum BDNF levels between patients, who required supplemental oxygen compare to other patients (A). BDNF serum levels in patients with CNS manifestations (B), dyspnea (C), and fever (D) with different requirement of supplemental oxygen. Significant differences were observed in serum BDNF levels between patients with CNS manifestations and dyspnea, who required supplemental oxygen compared to who do not required supplemental oxygen. ns not significant, *p < 0.05
COVID-19 is a systematic disease that frequently implies neurological and non-neurological manifestations, predominantly by inducing hypoxia. Brain-derived neurotrophic factor (BDNF) is a key factor in regulating functions of nervous and respiratory systems and has been strongly related to hypoxia. Therefore, this study planned to investigate BDNF association with the COVID-19 manifestations especially neurological impairments and the infection-induced hypoxia. We enrolled sixty-four COVID-19 patients and twenty-four healthy individuals in this study. Patients were divided into two groups, with and without neurological manifestations, and their serum BDNF levels were measured by enzyme-linked immunosorbent assay (ELISA). COVID-19 patients had significantly lower BDNF levels than healthy individuals (p = 0.023). BDNF levels were significantly lower in patients with neurological manifestations compared to healthy individuals (p = 0.010). However, we did not observe a statistically significant difference in BDNF levels between patients with and without neurological manifestations (p = 0.175). BDNF’s levels were significantly lower in patients with CNS manifestations (p = 0.039) and higher in patients with fever (p = 0.03) and dyspnea (p = 0.006). Secondly, BDNF levels have a significant negative association with oxygen therapy requirement (p = 0.015). These results strongly suggest the critical association between dysregulated BDNF and hypoxia in promoting COVID-19 manifestations, particularly neurological impairments.
Parkinson’s disease (PD) is the second most common neurodegenerative disorder. Immune mechanisms play an important role in the development of PD. The purpose of this study was to identify potential differentially expressed immune-related genes (IRGs), signaling pathways, and drugs in PD, which may provide new diagnostic markers and therapeutic targets for PD. Differentially expressed genes (DEGs) and IRGs were respectively obtained from the Gene Expression Omnibus (GEO) dataset and the ImmPort database. Weighted gene co-expression network analysis (WGCNA) was utilized to further identify hub IRGs. Core IRGs were obtained by intersection of DEGs and hub genes in the module of WGCNA, followed by construction of diagnostic models and regulation network establishment of long non-coding RNAs (lncRNAs)-miRNAs-diagnostic IRGs. Analysis of functional enrichment and protein–protein interaction (PPI) network and identification of related drugs of DEGs was performed. LILRB3 and CSF3R were identified as potential diagnostic markers for PD. Two regulatory pairs were identified based on LILRB3 and CSF3R, including XIST-hsa-miR-214-3p/hsa-miR-761-LILRB3 and XIST-hsa-miR-485-5p/hsa-miR-654-5p-CSF3R. LEP and IL1A were drug targets of Olanzapine. MMP9 and HSP90AB1 were drug targets of Bevacizumab. In addition, LEP and MMP9 were respectively drug targets of Lovastatin and Celecoxib. Herpes simplex infection (involved TNFRSF1A) and cytokine-cytokine receptor interaction (involved CSF3R, LEP, and IL1A) were the most remarkably enriched signaling pathways of DEGs. Identified IRGs and related signaling pathways may play critical roles in the development of PD. Additionally, LILRB3 and CSF3R can be considered as potential immune-related diagnostic markers for PD. LEP, IL1A, MMP9, and HSP90AB1 may be regarded as immune-related therapeutic targets for PD.
Propofol prevents erastin-induced cell injury: CCK-8 results: A HT-22 cells were treated with 0, 0.1, 0.25, 0.5, or 1 μM erastin for 8 h; B HT-22 cells were treated with 0, 2.5, 5, 10, 25, or 50 μM propofol in the presence of 0.5 erastin; or C HT-22 cells were treated with 25, 50, or 75 μM propofol. Cell viability was measured by a microplate reader after CCK-8 treatment. D, E Analysis of HT-22 cells on control, erastin (Era, 0.5 μM), Era + propofol (PPF, 50 μM), or Era + ferrostain-1 (Fer-1, 1 μM). D Microscopic examination. After the cells were treated with erastin, propofol, or ferrostain-1, the cells were observed by microscopy. Scale bar = 100 μm. E PI/Hoechst 33342 staining analysis. After erastin, propofol, or ferrostain-1 treatments, cells were stained with PI and Hoechst 33342 to identify the necrosis rate. *p < 0.05, **p < 0.01
Propofol prevents erastin-induced cell ferroptosis. Analysis of HT-22 cells treated with control, erastin (Era, 0.5 μM), Era + propofol (PPF, 50 μM), or Era + ferrostain-1 (Fer-1, 1 μM). A DHE-staining images and analysis. Cells were stained with DHE for 15 min and observed by a fluorescence microscope. Scale bar = 100 μm. B IF-staining images and analysis. Cells were fixed and immunostained with an anti-4-hydroxy-2- nonenal (4-HNE) antibody. Fluorescence signals were visualized by microscopy with constant fluorescence parameters. Scale bar = 100 μm. C Cells were stained with BODIPY C11 for 30 min to identify the lipid ROS level. The data were measured by a multifunction microplate reader. D FeRhoNox-staining images and analysis. Living cells were stained with FeRhoNox to identify the Fe²⁺ level, and cells were observed by fluorescence microscopy. Scale bar = 100 μm. E Representative EM image of HT-22 cells and quantification of injured mitochondria counts (red arrow indicates the injured mitochondria). Scale bar = 3 μm. F qRT-PCR was used to identify the PTGS2 gene fold in HT-22 cells. G, H Western blot results showing the protein (COX2, GPX4, xCT) levels in HT-22 cells. β-actin was used as an internal reference in whole-cell lysates. Each experiment was repeated 3 times. Data are expressed as the mean ± SEM. *p < 0.05, **p < 0.01
Propofol reduces p-ALOX5 to attenuate erastin-induced ferroptosis. Analysis of HT-22 cells on control, erastin (Era, 0.5 μM), or Era + propofol (PPF, 50 μM). A Western blot results showing the protein (ALOX5, p-ALOX5 (Ser 271, Ser 523, Ser 663)) level in HT-22 cells. β-actin was used as an internal reference in whole-cell lysates. B IF-staining images and analysis. Cells were fixed and immunostained with anti-p-ALOX5 (Ser 663) antibody. Fluorescence signals were visualized by microscopy with constant fluorescence parameters. Scale bar = 20 μm. C, D Western blot results showing the protein (MK2, p-MK2, PKA, p-PKA, ERK, p-ERK) levels in HT-22 cells. E Molecular docking showed propofol physically bound to the amino acid residue of ALOX5. β-actin was used as an internal reference in whole-cell lysates. Each experiment was repeated 3 times. Data are expressed as the mean ± SEM. *p < 0.05, **p < 0.01
Propofol protecting erastin-induced neuron injury is suppressed in Ser663Ala overexpressed HT-22 cells. Analysis of HT-22 cells on Ser663A OE + erastin (0.5 μM), Ser663A OE + erastin + propofol (50 μM), ALOX5 OE + erastin, or ALOX5 OE + erastin + propofol. A, B Western blot results showing the protein (ALOX5, p-ALOX5 (Ser 663)) level in HT-22 cells. β-actin was used as an internal reference in whole-cell lysates. C CCK-8 results: Cell viability was measured by a microplate reader after CCK-8 treatment. D FeRhoNox staining and analysis. Living cells were stained with FeRhoNox to identify the Fe²⁺ level, and cells were measured by a multifunction microplate reader. E Cells were stained with BODIPY C11 for 30 min to identify the lipid ROS level. The data were measured by a multifunction microplate reader. Each experiment was repeated 3 times. Data are expressed as the mean ± SEM. *p < 0.05, **p < 0.01
Propofol is a short-acting intravenous anesthetic that is widely used in clinical treatment. Previous articles have indicated that propofol is a therapeutic target for anti-apoptosis, anti-inflammation, anti-lipid peroxidation, and anti-reactive oxygen species (ROS). Moreover, cell ferroptosis is strongly correlated with cellular ROS, inflammatory responses, and lipid peroxidation. However, the mechanisms by which propofol attenuates neuronal injury by reducing ferroptosis remain unknown. Hence, we hypothesized that propofol could protect neurons by reducing ferroptosis. To test this hypothesis, HT-22 cells were treated with a specific ferroptosis activator (erastin) in the presence of propofol (50 μM). We found that propofol reduced erastin-induced high Fe²⁺ concentrations, lipid peroxides, and excess ROS. Western blotting results also suggested that propofol could rescue erastin-induced low expression of GXP4 and system Xc⁻. Further experiments indicated that propofol attenuated p-ALOX5 expression at Ser663 independent of ERK. In addition, we built two transient transfection cell lines, ALOX5 OE and Ser663Ala-ALOX5 OE, to confirm the target of propofol. We found that the Ser663 point is the critical role of propofol in rescuing erastin-induced cell injury/lipid peroxidation. In conclusion, propofol may help attenuate ferroptosis, which may provide a new therapeutic method to treat neuronal injury or the brain inflammatory response.
An increasing number of evidences have shown that the carcinogenic effect of DRAXIN plays an important role in the malignant process of tumors, but the mechanism of its involvement in glioma has not yet been revealed. The main aim of this study is to explore the relationship between DRAXIN and the prognosis and pathogenesis of glioma through a large quality of data analysis. Firstly, thousands of tissue samples with clinical information were collected based on various public databases. Then, a series of bioinformatics analyses were performed to mine data from information of glioma samples extracted from several reputable databases to reveal the key role of DRAXIN in glioma development and progression, with the confirmation of basic experiments. Our results showed that high expression of the oncogene DRAXIN in tumor tissue and cells could be used as an independent risk factor for poor prognosis in glioma patients and was strongly associated with clinical risk features. The reverse transcription-quantitative PCR technique was then utilized to validate the DRAXIN expression results we obtained. In addition, co-expression analysis identified, respectively, top 10 genes that were closely associated with DRAXIN positively or negatively. Finally, in vitro experiments demonstrated that knockdown of DRAXIN significantly inhibited proliferation and invasion of glioma cell. To sum up, this is the first report of DRAXIN being highly expressed in gliomas and leading to poor prognosis of glioma patients. DRAXIN may not only benefit to explore the pathogenesis of gliomas, but also serve as a novel biological target for the treatment of glioma.
The transporter hypothesis is one of the most popular hypotheses of drug-resistant epilepsy (DRE). P-glycoprotein (P-gp), a channel protein at the blood–brain barrier (BBB), plays an important role in the transport of some anti-seizure drugs from brain tissue into vessels, which reduces drug concentrations and diminishes the effects of drug treatment. We performed this study to test whether P-gp is overexpressed in DRE and identify ways to prevent and reverse DRE. In this study, we established a phenytoin (PHT)-resistant mouse model and revealed that P-gp was overexpressed at the BBB in PHT-resistant mice. The P-gp inhibitor nimodipine decreased the resistance of phenytoin. Antioxidative preventive treatment with N-acetylcysteine (NAC) prevented the mice from entering a PHT-resistant state, and NAC therapy tended to reverse PHT resistance into sensitivity. We were also able to induce PHT resistance by activating the Nrf2/P-gp pathway, which indicates that oxidative stress plays an important role in drug resistance. Taken together, these findings suggest that antioxidative therapy may be a promising strategy for overcoming DRE.
Brain regions with recognition of positive and negative music stimulation in participants with MDD
Brain regions with recognition of positive and negative emotional stimuli in ND subject. Combination means the intersection of the feature selection results of the first two algorithms
Brain regions with recognition of positive and negative music stimulation in participants with MDD (A) Mean FC was averaged across 15 × 14/2 = 105 connections. 15 ROIs were the ERBRs under positive and negative stimulation in MDD. (B–C) Mean FC matrix diagram of depression participants stimulated by pEMS (B) and nEMS (C). (D) FC differences under pEMS and nEMS in MDD. The positive FC matrix subtracted the difference values from the negative FC matrix. Color bars correspond to the FC values. (E) The FC between Cingulum_Mid_L and Temporal_Sup_R showed a significant difference between positive verse negative stimuli. FC_i-j symbolizes FC between ROI_i and ROI_j. The numerical labelling of the brain area corresponds to the description document of the AAL template
(A–B) Mean FC matrix diagram of ND under pEMS (A) and nEMS (B). 15 ROIs were the ERBRs under positive and negative stimulation in MDD. (C) FC differences under pEMS and nEMS in ND. The positive FC matrix subtracted the difference values from the negative FC matrix. Color bars correspond to the FC values. (D) FC network diagram with significant difference between positive and negative stimuli in ND group. (E) The box plot of FC distribution with a significant difference. FC_i-j symbolizes FC between ROI_i and ROI_j. The numerical labelling of the brain area corresponds to the description document of the AAL template
(A) Mean FC was averaged across 14 × 13/2 = 91 connections. 14 ROIs were the ERBRs under positive and negative stimulation in ND. (B) The FCs with significant difference between positive verse negative stimuli for ND participants. (C and E) FC network diagrams with significant differences between NDs and MDDs under pEMS (C) and nEMS (E). (D and F) The box plot of FC distribution with a significant difference between MDDs and NDs under pEMS (D) and nEMS (F)
Depression is characterized by poor emotion regulation that makes it difficult to escape the effects of emotional pain, but the neuromodulation behind these symptoms is still unclear. This study investigated the neural mechanism of emotional state-related responses during music stimuli in participants with major depressive disorder (MDD) compared to never-depressed (ND) controls. A novel two-level feature selection method, integrating recursive feature elimination based on support vector machine (SVM-RFE) and random forest algorithm (RF), was proposed to screen emotional recognition brain regions (ERBRs). On this basis, the differences of functional connectivity (FC) were systematically analyzed by two-sample t-test. The results demonstrate that ND participants show eight pairs of FCs with a significant difference between positive emotional music stimuli (pEMS) versus negative emotional music stimuli (nEMS) in 15 ERBRs of MDD, but the participants with MDD show one pair of significant difference in FC. The decreased number reflects the fuzzy response to positive and negative emotions in MDD, which appears to arise from obstacle to emotional cognition and regulation. Furthermore, there was no significant difference in FC between MDDs and NDs under pEMS, but a significant difference was detected between the two groups under nEMS (p < 0.01), revealing a ‘bias’ against the negative state in MDD. The current study may help to better comprehend the abnormal evolution from normal to depression and inform the utilization of pEMS in formal treatment for depression.
Previous reports have confirmed the significance of CD93 in the progression of multiple tumors; however, there are few studies examining its immune properties for gliomas. Here, we methodically investigated the pathophysiological characteristics and clinical manifestations of gliomas. Six hundred ninety-nine glioma patients in TCGA along with 325 glioma patients in CGGA were correspondingly collected for training and validating. We analyzed and visualized total statistics using RStudio. One-way ANOVA and Student’s t -test were used to assess groups’ differences. All differences were considered statistically significant at the level of P < 0.05. CD93 markedly upregulated among HGG, MGMT promoter unmethylated subforms, IDH wild forms, 1p19q non-codeletion subforms, and mesenchyme type gliomas. ROC analysis illustrated the favorable applicability of CD93 in estimating mesenchyme subform. Kaplan–Meier curves together with multivariable Cox analyses upon survivance identified high-expression CD93 as a distinct prognostic variable for glioma patients. GO analysis of CD93 documented its predominant part in glioma-related immunobiological processes and inflammation responses. We examined the associations of CD93 with immune-related meta-genes, and CD93 positively correlated with HCK, LCK, MHC I, MHC II, STAT1 and IFN, while adverse with IgG. Association analyses between CD93 and gliomas-infiltrating immunocytes indicated that the infiltrating degrees of most immunocytes exhibited positive correlations with CD93, particularly these immunosuppressive subsets such as TAM, Treg, and MDSCs. CD93 is markedly associated with adverse pathology types, unfavorable survival, and immunosuppressive immunocytes infiltration among gliomas, thus identifying CD93 as a practicable marker and a promising target for glioma-based precise diagnosis and therapeutic strategies.
In multiple sclerosis patients, long-term inflammation makes the oligodendrocyte progenitor cells (OPCs) exhausted; therefore, a new therapy that makes them responsive to insults to participate in remyelination is highly in demand. Here, we investigated the effect of ursolic acid (UA) on myelin repair after mid-term and long-term demyelination periods induced by 6 or 12 weeks of cuprizone treatment followed by 2 weeks of recovery with or without UA. Immunohistochemistry studies and myelin genes expression assessment were used to evaluate the myelination status of mouse corpora callosa and the cellular mechanisms of myelin repair. Results showed that UA significantly promoted recovery from myelin loss after discontinuing 6 or 12 weeks of cuprizone feeding, as measured by luxol fast blue (LFB), fluoroMyelin (FM), anti-myelin basic protein (MBP) staining, and oligodendrocyte progenitor cell counts. It led to reduced inflammation and gliosis as evaluated by glial fibrillary acidic protein (GFAP), Iba1, or other marker gene transcripts. Following long-term demyelination, gliosis and TNF-α were observed as potential players in lesion pathology, which were restored by UA. An increased IL-10 may contribute to UA anti-inflammatory effect and making responsive the exhausted OPCs. UA increased the number of new oligodendrocyte lineage cells and myelination. Our findings indicated that UA can enhance myelin repair after cuprizone challenge through the prevention of gliosis and increasing the newly generated myelin.
We recently showed that membrane progesterone receptor α (mPRα/PAQR7) promotes pro-regenerative effects in Schwann cell-like adipose stem cells (SCL-ASC), an alternative model to Schwann cells for the promotion of peripheral nerve regeneration. In this study, we investigated how mPRα activation with the mPR-specific agonist Org OD 02–0 in SCL-ASC affected regenerative parameters in two neuronal cell lines, IMR-32 and SH-SY-5Y. In a series of conditioned medium experiments, we found that mPR activation of SCL-ASC led to increased neurite outgrowth, protection from cell death and increased expression of peripheral nerve regeneration markers (CREB3, ATF3, GAP43) in neuronal cell lines. These effects were stronger than the ones observed with the conditioned medium from untreated SCL-ASC. The addition of Org OD 02–0 to the untreated cell medium mimicked the effects of mPR activation of SCL-ASC on cell death, but not on neurite outgrowth. Therefore, the effect of Org OD 02–0 on neurite outgrowth is SCL-ASC-dependent, while its effect on cell survivability is likely due to the direct activation of mPRs on neuronal cells. SCL-ASC transfection with mPRα siRNA showed that this isoform is responsible for the beneficial effect on neurite outgrowth. Further experiments showed that SCL-ASC-dependent outcomes likely involved the release of BDNF and IGF-2 from these cells. The beneficial mPRα effect on neurite outgrowth was confirmed in co-culture conditions. These findings strengthen the hypothesis that mPRα could play a pro-regenerative role in SCL-ASC and be a therapeutic target for the promotion of peripheral nerve regeneration.
Average standard scores (SS). This figure shows the average standard scores (SS) of the 4 tested patients. All results were below the 1 percentile rank of the age-matched general population, except for motor skills for subject B at the age 6.6 (6 years and 6 months) when he was at a 2 percentile rank of the general population. Please note, the age numbers before the decimal points are given in years and after the decimal point the numbers are given in months
Age equivalent (AE) scores indicating the age level at which the subject's raw score is found at a norm sample median. The figure compares AE of Subject A to B during development as well as for adults (C and D) as in Fig. 1. Please note that the different subdomain trait measures cannot be compared. Furthermore, when the AE depicted is a round number such as 2 or 3 years, the actual measurement was actually small than 2 or 3 years. Lastly, as in Fig. 1, the numbers before the decimal points are given in years and after the decimal point the number are given in months
Growth scale values (GSV) comparing Subject A to B during development. GSV quantities change for subject A shows mild improvements especially in interpersonal, play and leisure, and gross motor subdomains. For subject B the graph shows minor improvements in the various subdomains
Activity-dependent neuroprotective protein (ADNP) is one of the lead genes in autism spectrum disorder/intellectual disability. Heterozygous, de novo ADNP mutations cause the ADNP syndrome. Here, to evaluate natural history of the syndrome, mothers of two ADNP syndrome boys aged 6 and a half and two adults aged 27 years (man and woman) were subjected to Vineland III questionnaire assessing adaptive behavior. The boys were assessed again about 2 years after the first measurements. The skill measures, presented as standard scores (SS) included domains of communication, daily living, socialization, motor skills and a sum of adaptive behavior composite. The age equivalent (AE) and growth scale values (GSV) encompassing 11 subdomains assess the age level at which the subject’s raw score is found at a norm sample median and the individual temporal progression, respectively. The norm referenced standard scores age-matched, mean 100 ± 15 of the two children showed the lowest outcome in communication (SS: 20–30). Daily living skills presented SS of 50–60, with a possible potential loss of some activities as the child ages, especially in interpersonal relationships with people outside of the immediate family (boy A). In contrast, in socialization, both children were at the SS of 38, with some positive increase to SS of ~ 45 (interpersonal relations with family members and coping skills, depending on the particular individual), 2 years later. Interestingly, there was an apparent large difference in motor skills (gross and fine) at the young age, with subject B showing a relatively higher level of skills (SS: 70), decreasing to subject A level (SS: 40) 2 years later. Together, the adaptive behavior composite suggested a level of SS: 39–48 with B showing a potential increase (SS: 41–44) and A, a substantial decrease (SS: 48–39), suggesting a strong impact of daily living skills. Adults were at SS: 20, which is the lowest possible score. AE showed minor improvements for subject A and B, with all AE values being below 3 years. GSVs for subject A showed some improvement with age, especially in interpersonal, play and leisure, and gross motor subdomains. GSV for subject B showed minor improvements in the various subdomains. Notably, all subjects showed a percentile rank < 1 compared with age-matched norms except for subject B as to motor domain (2nd percentile) at the age of 6 years. In summary, the results, especially comparing SS and AEs between childhood and adulthood, implied a continuous deterioration of activities compared to the general population, encompassing a slower developmental process coupled to possible neurodegeneration, strongly supporting a great need for disease modifying medicinal procedures.
Different interneuron classes have distinct laminar distribution patterns which contribute to the layer-specific organization of cortical microcircuits. However, laminar differences within the same interneuron classes are not well recognized. Despite systematic efforts towards neuron cell-type taxonomy in the neocortex by single-cell transcriptomics, less attention has been driven towards laminar differences in interneurons compared to projection neurons. VIP⁺ interneurons are the major interneuron class that mostly populate superficial layers and mediate disinhibition. A few reports noted the morphological and electrophysiological differences between VIP⁺ interneurons residing in layers I–III (upper layer) and layers IV–VI (deeper layer), but little is known about their molecular differences. Here, we delineated the laminar difference in their transcriptome employing single-cell RNA sequencing (scRNAseq) data from public databases. Analysis of 1175 high-quality VIP⁺ interneurons in the primary visual cortex (VISp) showed that the upper layer and deeper layer VIP⁺ interneurons are transcriptionally distinct distinguished by genes implicated in synapse organization and regulation of membrane potential. Similar differences are also observed in the anterior lateral motor cortex (ALM) and primary motor cortex (MOp). Cross-comparing between the top 10 differentially expressed genes (DEGs) with Allen Mouse Brain in situ hybridization database, we identified Tac2 and CxCl14 as potential marker genes of upper layer VIP⁺ interneurons across most cortical regions. Importantly, such expression patterns are conserved in the human brain. Together, we revealed significant laminar differences in transcriptomic profiles within VIP⁺ interneurons, which provided new insight into their molecular heterogeneity that may contribute to their functional diversity.
The P2X4 receptor (P2X4R) can be upregulated after nerve injury, and its mediated spinal microglial activation makes a critical contribution to pathologically enhanced pain processing in the dorsal horn. Although some studies have partly clarified the mechanism underlying altered P2X4R expression, the specific mechanism is not well understood. MicroRNAs (miRNAs) are small noncoding RNAs which control gene expression by binding with their target mRNAs. Thus, in the present study, we investigated whether miRNA is involved in the pathogenesis of neuropathic pain by regulating P2X4R. Our results showed that P2X4R was upregulated in the spinal dorsal horn of mice following spared nerve injury (SNI), and 69 miRNAs (46 upregulated and 23 downregulated miRNAs) were differentially expressed (fold change > 2.0, P < 0.05). P2X4R was found to be a major target of miR-106b-5p (one of the downregulated miRNAs) using bioinformatics technology; quantitative real-time PCR analysis confirmed the change in expression of miR-106b-5p, and dual-luciferase reporter assays confirmed the correlation between them. Fluorescence in situ hybridization was used to show cell co-localization of P2X4R and miR-106b-5p in the spinal dorsal horn. Transfection with miR-106b-5p mimic into BV2 cells reversed the upregulation of P2X4R induced by lipopolysaccharide (LPS). Moreover, miR-106b-5p overexpression significantly attenuated neuropathic pain induced by SNI, with decreased expression of P2X4R mRNA and protein in the spinal dorsal horn; intrathecal miR-106b-5p antagomir induced pain behaviors, and increased expression of P2X4R in the spinal dorsal horn of naïve mice. These data suggest that miR-106b-5p can serve as an important regulator of neuropathic pain development by targeting P2X4R.
Imaging genetics using imaging technology is regarded as a neuroanatomical phenotype to evaluate gene single nucleotide polymorphisms and their effects on the structure and function of different brain regions. It plays a vital role in bridging the initial understanding of the genetic basis of brain structure and dysfunction. Sparse canonical correlation analysis (SCCA) has become a widespread technique in this field because of its powerful ability to identify bivariate relationships and feature selection. Since most traditional SCCA algorithms assume that the input features are independent, this method obviously cannot be used to analyze genetic image data. The MT-SCCA model is unsupervised and cannot identify the genotype–phenotype associations for diagnostic guidance. Meanwhile, a single biological clinical index cannot fully reflect the physiological process of a comprehensive disease. Therefore, it is necessary to find biomarkers that can reflect Alzheimer’s disease and physiological functions that can more comprehensively reflect the development of the disease. This article uses a multi-task sparse canonical correlation analysis and regression (MT-SCCAR) model to combine the annual depression level total score (GDSCALE), clinical dementia assessment scale (GLOBAL CDR), functional activity questionnaire (FAQ), and neuropsychiatric Symptom Questionnaire (NPI-Q) in this paper. These four clinical data are used as compensation information and embedded in the algorithm in a linear regression manner. It also reflects its superiority and robustness compared to traditional correlation analysis methods on actual and simulated data. Meanwhile, compared with MT-SCCA, the model utilized in this paper obtains a higher gene-ROI weight and identifies clearer biomarkers, which provides a practical basis for the study of complex human disease pathology.
Spinal cord injury (SCI) can cause permanent neurological deficits. Circular RNA Ncam2 (circ-Ncam2 also termed mmu_circ_0006413) has been reported to be overexpressed in SCI mouse models. However, the function of circ-Ncam2 in SCI has not been validated. Lipopolysaccharide (LPS) was used to activate mouse microglia (BV2 cells). Expression levels of circ-Ncam2 were determined by RT-qPCR. Relative protein levels were evaluated by western blotting. Cytokines were determined by ELISA. The regulatory mechanism of circ-Ncam2 was validated by dual-luciferase reporter and RNA pull-down assays. Effects of LPS-induced BV2 cells on mouse neuronal (HT22 cells) viability, proliferation, and apoptosis were analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide (MTT), 5-ethynyl-2′-deoxyuridine (EdU), and flow cytometry assays. LPS stimulation promoted circ-Ncam2 expression in BV2 cells. Inhibition of circ-Ncam2 mitigated LPS-induced BV2 cell activation and inflammation. Mechanically, circ-Ncam2 adsorbed miR-544-3p to regulate TLR4 expression. Also, either miR-544-3p inhibition or TLR4 overexpression weakened circ-Ncam2 silencing-mediated effects on LPS-induced BV2 cell activation and inflammation. Furthermore, LPS-induced BV2 cells suppressed HT22 cell proliferation and promoted HT22 cell apoptosis through the circ-Ncam2/miR-544-3p axis. Importantly, circ-Ncam2 activated the NF-κB signaling via the miR-544-3p/TLR4axis. circ-Ncam2 silencing lowered LPS-induced microglia activation and neuronal apoptosis via blocking the TLR4/NF-κB pathway through acting as a miR-544-3p sponge, suggesting that circ-Ncam2 may be involved in secondary SCI.
Flowchart of the study design. The figure depicts the outline of the study model
Kaplan–Meier curve illustrating cholesterol parameters in three PD age categories which depicts the probability (P < 0.05) of metabolites at risk after PD diagnosis. The curve represents the significance of LO and EO groups at P < 0.05 who are at survival risk with cholesterol metabolites. LO, late onset; EO, early onset; J, juvenile
Kaplan–Meier curve illustrating kynurenine parameters in three PD age categories which depicts the probability (P < 0.05) of metabolites at risk after PD diagnosis. The curve represents the significance of LO and EO groups at P < 0.05 who are at survival risk with kynurenine metabolites. LO, late onset; EO, early onset; J, juvenile
Parkinson’s disease (PD) is an ageing disorder caused by dopaminergic neuron depletion with age. Growing research in the field of metabolomics is expected to play a major role in PD diagnosis, prognosis and therapeutic development. In this study, we looked at how SNCA and GBA1 gene mutations, as well as metabolomic abnormalities of kynurenine and cholesterol metabolites, were linked to alpha-synuclein (α-syn) and clinical characteristics in three different PD age groups. In all three age groups, a metabolomics analysis revealed an increased amount of 27-hydroxycholesterol (27-OHC) and a lower level of kynurenic acid (KYNA). The effect of 27-OHC on SNCA and GBA1 modifications was shown to be significant (P < 0.05) only in the A53T variant of the SNCA gene in late-onset and early-onset PD groups, whereas GBA1 variants were not. Based on the findings, we observed that the increase in 27-OHC would have elevated α-syn expression, which triggered the changes in the SNCA gene but not in the GBA1 gene. Missense variations in the SNCA and GBA1 genes were investigated using the sequencing technique. SNCA mutation A53T has been linked to increased PD symptoms, but there is no phenotypic link between GBA1 and PD. As a result of the data, we hypothesise that cholesterol and kynurenine metabolites play an important role in PD, with the metabolite 27-OHC potentially serving as a PD biomarker. These findings will aid in the investigation of pathogenic causes as well as the development of therapeutic and preventative measures for PD.
Unique SETX variants in two families with ataxia-oculomotor apraxia 2 (AOA2). A Pedigree of Family 1, showing segregation of the in-frame deletion, and duplication variants; B Pedigree of Family 2, showing segregation of the nonsense, and intronic variants; C MRI T1 proband II-4 Family 1, sagittal, and showing atrophy of vermis; D MRI T1 proband II-4 Family 1, para sagittal, and showing cerebellar hemispherical atrophy; E MRI flair proband II-3 Family 2, sagittal, and showing atrophy of vermis; F MRI flair proband II-3 Family 2, para sagittal, and showing mild cerebellar upper hemispherical atrophy; G Schematic representation of the senataxin protein and its active domains. The locations of variants observed in both families are marked
Functional and structural analyses of the SETX variants in family 1. A Induced chromosomal aberrations (ICA) scores per cell after exposure to different concentrations of mitomycin-C (MMC) in patients compared to control revealed that AOA2 patients show higher chromosomal breakage rates at baseline and 200 nM MMC; B Cell viability at 72 h post MMC treatment at different concentrations reveals that AOA2 patient LCLs show significantly higher sensitivity (i.e. lower viability) to 15 µM MMC compared to control LCLs (p = 0.037), which reaches a plateau despite exposure to a higher MMC concentration (30 µM); C 3D structure of the senataxin helicase domain (blue) shows that Val2337 (red) is buried and forms hydrogen bonds (pink) with Lys2331 and Arg2334, while His2457 (red) is exposed to the solvent and located right outside the helicase domain
RNA analysis of intronic variant in Family 2. A Gel electrophoresis reveals that the paternally-inherited c.5549-107A > G variant leads to abnormal alternative splicing and produces an allele with higher molecular weight cDNA product than the WT allele; B–C Sanger sequencing using mis-spliced allele-specific primers reveals that c.5549-107A > G induces activation of cryptic exon 12a, leading to an insertion of 73 bp from intron 12 which contain a premature stop codon; D RT-qPCR analysis reveals that while in WT allele state, there is naturally very low transcription of cryptic exon 12a (mother and control), the A-to-G substitution leads to 20–30 times higher transcription rates of the mis-spliced mRNA (patient 2 and father); E RT-qPCR analysis of total SETX mRNA reveals that the combination of the paternal intronic variant with the maternal nonsense variant leads to lower SETX mRNA levels and disease manifestations in the patient
AOA2 is a rare progressive adolescent-onset disease characterised by cerebellar vermis atrophy, peripheral neuropathy and elevated serum alpha-fetoprotein (AFP) caused by pathogenic bi-allelic variants in SETX, encoding senataxin, involved in DNA repair and RNA maturation. Sanger sequencing of genomic DNA, co-segregation and oxidative stress functional studies were performed in Family 1. Trio whole-exome sequencing (WES), followed by SETX RNA and qRT-PCR analysis, were performed in Family 2. Sanger sequencing in Family 1 revealed two novel in-frame SETX deletion and duplication variants in trans (c.7009_7011del; p.Val2337del and c.7369_7371dup; p.His2457dup). Patients had increased induced chromosomal aberrations at baseline and following exposure to higher mitomycin-C concentration and increased sensitivity to oxidative stress at the lower mitomycin-C concentration in cell viability test. Trio WES in Family 2 revealed two novel SETX variants in trans, a nonsense variant (c.568C > T; p.Gln190*), and a deep intronic variant (c.5549-107A > G). Intronic variant analysis and SETX mRNA expression revealed activation of a cryptic exon introducing a premature stop codon (p.Met1850Lysfs*18) and resulting in aberrant splicing, as shown by qRT-PCR analysis, thus leading to higher levels of cryptic exon activation. Along with a second deleterious allele, this variant leads to low levels of SETX mRNA and disease manifestations. Our report expands the phenotypic spectrum of AOA2. Results provide initial support for the hypomorphic nature of the novel in-frame deletion and duplication variants in Family 1. Deep-intronic variant analysis of Family 2 variants potentially reveals a previously undescribed poison exon in the SETX gene, which may contribute to tailored therapy development.
LRP1 expression in epilepsy patients and normal people. A The count of LRP1 gene showed no difference between normal people (control) and epilepsy patients in GSE139914 database (Brodmann area 38). B The LRP1 mean count of epilepsy patients is lower than that of normal people in GSE134697 database (neocortex)
Knockdown of LRP1 induces seizure-like behavior in Drosophila. A Seizure behavior in flies (Black arrow). B Seizures occurred at a higher rate in Lrp1 knockdown flies (tub-Gal4 > Lrp1-RNAi) than in WT flies (Canton-S) and other control groups (tub-Gal4, Lrp1-RNAi). The tub-Gal4 > Scn1a-RNAi positive controls had a higher rate of seizures than the tub-Gal4 > Lrp1-RNAi group. C Ketone body diet largely inhibited the seizure rate in Lrp1 global knockdown flies. High sucrose could increase the seizure rate in WT flies. Data were given as mean ± S.D. *P < 0.05, ***P < 0.005, and ****P < 0.001 in comparison between each two groups in Tukey’s post-hoc test and in comparison among multiple samples in One-way ANOVA
LRP1 defect induced partial loss of central complex. A–C Serial images of the brain of WT flies from anterior to posterior show the structure of the central complex including ellipsoidal body (A), fan-shaped body (surf) (B), and fan-shaped body (deep) (C). D–F Serial images of the brain of a Lrp1 knockdown fly from anterior to posterior showing the structure of the ellipsoidal body (D), fan-shaped body (surf) (E), and fan-shaped body (deep) (F). Scale bar 50 µm. The hollow arrows indicated the relevant brain structures. The solid arrows indicated the abnormal structure in the central complex
Pan-glial knockdown of LRP1 sufficiently induced seizure behavior. A Seizures occurred at a higher rate in Lrp1 global knockdown flies tub-Gal4 > Lrp1-RNAi than in Lrp1 pan-glial knockdown flies repo-Gal4 > Lrp1-RNAi. The repo-Gal4 > Lrp1-RNAi flies had a higher rate of seizures than of the pan-neural knockdown flies elva-Gal4 > Lrp1-RNAi group. B The pan-neural Lrp1 knockdown flies elav-Gal4 > Lrp1-RNAi fed with high sucrose food had a higher rate of seizures than that feeding standard food. Ketone body diet completely inhibited the seizure of those flies. C Seizures occurred at a higher rate in glial knockdown flies repo-Gal4 > Lrp1-RNAi fed with standard food than that feeding for ketone body diet. Data were given as mean ± S.D. *P < 0.05, **P < 0.01, ***P < 0.005, and ****P < 0.001 in comparison between each two groups in Tukey’s post-hoc test and in comparison among multiple samples in One-way ANOVA
Ketone body diet rescued LRP1 deficiency by downregulating glutamate transmission. A Knockdown of glial glutamate transport (repo > Eaat1-RNAi) induces serious seizure behavior, and ketone body diet could not inhibit the seizure rate in glial glutamate transporter knockdown flies. B Over-expression of Eaat1 could inhibit seizure caused by LRP1 deficiency. ST is short for the standard diet; KB is short for the ketone body diet. Data were given as mean ± S.D. *P < 0.05, **P < 0.01, and ***P = 0.005 in comparison between each two groups in Tukey’s post-hoc test and in comparison among multiple samples in One-way ANOVA
LRP1, the low-density lipoprotein receptor 1, would be a novel candidate gene of epilepsy according to our bioinformatic results and the animal study. In this study, we explored the role of LRP1 in epilepsy and whether beta-hydroxybutyrate, the principal ketone body of the ketogenic diet, can treat epilepsy caused by LRP1 deficiency in drosophila. UAS/GAL4 system was used to establish different genotype models. Flies were given standard, high-sucrose, and ketone body food randomly. The bang-sensitive test was performed on flies and seizure-like behavior was assessed. In morphologic experiments, we found that LRP1 deficiency caused partial loss of the ellipsoidal body and partial destruction of the fan-shaped body. Whole-body and glia LRP1 defect flies had a higher seizure rate compared to the control group. Ketone body decreased the seizure rate in behavior test in all LRP1 defect flies, compared to standard and high sucrose diet. Overexpression of glutamate transporter gene Eaat1 could mimic the ketone body effect on LRP1 deficiency flies. This study demonstrated that LRP1 defect globally or in glial cells or neurons could induce epilepsy in drosophila. The ketone body efficaciously rescued epilepsy caused by LRP1 knockdown. The results support screening for LRP1 mutations as discriminating conduct for individuals who require clinical attention and further clarify the mechanism of the ketogenic diet in epilepsy, which could help epilepsy patients make a precise treatment case by case.
Top-cited authors
Dora Reglodi
  • University of Pécs
Daniel Offen
  • Tel Aviv University
Eldad Melamed
  • Tel Aviv University
Andrea Tamas
  • University of Pécs
Yunhui Liu