Questions related to Neuroinflammation
Recently, I was introduced to perivascular spaces (PVS) while studying lipopolysaccharide treated models with transmission electron microscopy (TEM).
Erickson MA, Shulyatnikova T, Banks WA, Hayden MR. Ultrastructural Remodeling of the Blood-Brain Barrier and Neurovascular Unit by Lipopolysaccharide-Induced Neuroinflammation. Int J Mol Sci. 2023; 23(24(2). DOI:10.3390/ijms24021640
PVS fascinates and intrigue me. I know that they are normally present in post-capillary venules and that they are a biomarker for small vessel disease (SVD) they are enlarged (EPVS) and EPVS are increased in aging.
I look forward to any input and suggestions for me as I will soon begin studying very old CD-a mice brains and I am in hopes to find some new knowledge regarding EPVS with the use of TEM.
Please make suggestions and submit any papers that you feel might be important in this future study.
Melvin R Hayden
University of Missouri School of Medicine
TEM core at the NexGen Precision Health Research Center
Columbia, Missouri USA
I am trying to find a mouse astrocyte cell line. I was planning to order C8-D1A from ATTC for some neuroinflammation experiments but the website was recently banned in my country (central Europe). I am grateful for any recommendation. Thank you.
The method I am looking for should be non-invasive and within the scope of MRI modalities.
Is lipocalin-2 a good guy during neuroinflammation?
There are contrasting reports regarding this protein. Some say it protects the integrity of the BBB during inflammatory conditions other researchers say otherwise.
What do you think?
Let's share some ideas
I know this method published in PLoS ONE but do not whether I will be able to use it as I do not have matlab. I will try to use ImageJ or FIJI...
Kozlowski C, Weimer RM (2012) An Automated Method to Quantify Microglia Morphology and Application to Monitor Activation State Longitudinally In Vivo. PLoS ONE 7(2): e31814. doi:10.1371/journal.pone.0031814
Thanks in advance for your help.
I recently had a video created from one of my manuscripts in Neuroglia MDPI entitled:
Hypothesis: Neuroglia Activation Due to Increased Peripheral and CNS Proinflammatory Cytokines/Chemokines with Neuroinflammation May Result in Long COVID. Neuroglia 2021, 2(1), 7-35; https://doi.org/10.3390/neuroglia2010004
Here is the video link:
This video runs about 10 minutes and pretty much tells the story of the manuscript in a video highlighted fashion.
In today's fast-moving internet society and the younger population wanting things condensed for them and increasing use of smartphones all around us the utilization of videos regarding scientific manuscripts might become popular. While the manuscript remains available online and most are available for open free access it might be possible that videos might increase the readership of the full manuscript and that the videos will be sort of like an introduction to invite more readers for viewing the author's work? This is open now for discussion.
Melvin R Hayden
Do you have any experience in preparing sample dilutions to measure the concentrations of inflammatory cytokines (TNFα, TGFβ, MCP-1, IL-1α, IL-1β, IL-6, and so forth) for an ELISA when it comes to any rat brain tissue, especially hippocampus. I'd prefer not to perform a pretest, and waste any well-strips beforehand. Based on your experience from different labs, what would be the expected protein concentration ranges of these cytokines in a supernatant saved from tissue homogenate prepared from a healthy rat (Sprague Dawley) brain tissue (hippocampus)?
I would like to ask if we can apply the term "Immunoreactivity" within the spinal cord or within the brain as the alternative keyword to study about Neuroinflammation? And how is the correlation between them?
And is every immunoreactivity tissue sampling method can be included as Neuroinflammation process too?
Thank You in Advanced.
A peer-reviewed journal, Neuroimmunology and Neuroinflammation opened a special issue to publish review articles about recent advances on Parkinson's disease. Please refer the link above to have more information how to submit it. Thanks.
I am currently studying a proteolytically stable peptide that transiently increases blood brain barrier permeability. While the results suggest that it could facilitate drug delivery to the brain, we are interested in assessing the potential downside of such a strategy.
Primarily, we are worried about neuroinflammation. My lab does not have the facilities to properly detect neuroinflammation. Does anyone know of any lab, core facility, or private company that offers services that can determine whether or not the peptide we are testing can lead to neuroinflammation?
I want to ask about CNS in the Neuroinflammation/Neuroimmunology concept.
Is it correct to say that CNS is defined as immune privilege organ due to its unique of immune reaction compare with the general immune system? Is the unique refers to the ability of CNS to induce its own immune reaction, independent from other peripheral immunity, or any other reasons?
And is it still updated to classify CNS as immune privileged system?
Thanks in advance for your answers and comments
I am using an organotypic hippocampal slice culture where we are exposing our slices to alcohol with subsequent alcohol withdrawal.
I want to make sure I am capturing the right time to assess TNFa concentrations. We are currently collecting and assessing 24 hours after the treatment, but I'm concerned that TNFa concentrations may be more transient (i.e. TNFa may act quickly and be subsequently degraded in a window of time that I'm missing at 24 hours compared to something like 30 minutes or an hour after treatment).
Does anyone have any suggestions/can anyone direct me to a reference regarding the optimal window of time for assessing TNFa concentrations?
ELISA details: analytical sensitivity of <4 pg/mL and an assay range of 11.7-750 pg/mL
my lab is in São Paulo, Brazil. There is a postdoc position open for the periodo of 12 month, starting in 2020. The plan is to study neuroinflammation caused by SARS-CoV-2 infection in bioprinted neural tissue.
Besides the damages to the respiratory system, studies indicate that the central nervous system is also affected in severe cases of COVID-19. The main symptoms are headaches, seizures, and consciousness disturbance, and necropsies revealed that infected patients show neural degeneration. Infection can result in severe neurological complications such as viral encephalitis, toxic encephalitis, and acute cerebrovascular diseases. The working plan proposes the production of SARS-CoV-2 in a bioprinted neural tissue, followed by the analysis of neuroinflammatory response.
Experience required in 3D bioprinting or brain organoids or 3D culture or culture and differentiation of h-iPSC in neural cells. Knowledge of molecular biology techniques (qPCR, RNAseq), and cellular analysis (immunocytochemistry, flow cytometry, live-cell microscopy) is desirable.
To apply, email me your CV, a letter of motivation and two reference letters. Pre-selected candidates will be invited to a conference call.
I am trying to isolate lymphocytes and/or myeloid cells for FACS analysis from the cohlea of mice p.n. day 4-16. Considering that there is a lot of hematopoietic bone marrow in the temporal bone itself, even though I microdissect the tissue, I get a lot of hematopoietic cells which, naturally, interferes with my data. Does anyone have any experience with this?
I've been trying various iba1 antibodies with either Alexa 488 or 647 and am getting a lot of background and have never once seen anything close to resembling a microglia. I've tried blocking with various combinations of 0, .3 or 3.0% milk, 0, .3 or 1.0% BSA, with 4% normal donkey serum and nothing has worked.
Any suggestions would be very much appreciated. Perhaps there are better membrane-bound proteins I could stain for?
According to researches, appendix might has connection with Parkinson's Disease but consequences are conflict. One study says, appendix removal reduces risk of PD but another study says that it increases.
Uncovering a Link Between the Appendix and Parkinson Disease Risk
I've been on neuronal cell culture to study neuroinflammation, but there has been a debate on neuronal cells not being relevance or giving significant role in neuroinflammation. It should have been immune cells like microglial cells.
But what if I want to proceed with neuronal cells, what are the markers that can be used other than inflammatory cytokine and chemokine?
how long does the cerebral cognitive impairment following LPS administration last? if a doses of 250ug/kg/day of LPS is used 055:B5
I want to know that whether there is any agent which can suppress the neuroinflammation in brain or not?
In brain, neuroinflammation occurs mainly because of the microglia or astrocytes. So, minocycline can reduce the microglia mediated inflammtion but I want to know something which can reduce the inflammtion cause by both cells i.e. astrocyte and microglia
I want to study the demyelination degree of EAE mice. I know that in the EAE model, lesions are preferentially located in the spinal cord and due to that it is very common to use LFB staining for spinal cord. Nevertheless I´m also interested in analyzing the brain. Has anyone used this staining in brain frozen tissue?
Thank you very much in advice
PTSD is associated with inflammation but the cause is unclear. What do you think?. Does neuroinflammation cause the symptoms of PTSD or does PTSD cause neuroinflammation and systemic inflammation?
I am working on a combined model of chronic immune activation via LPS and chronic mild stress. There is a discripancy in behavioral and cognitive changes between CMS vs.LPS/CMS and I wonder how LPS exposure could modulate response to CMS
Can peripheral blood be used to check neuroinflammation or microglial activation? or Do we have any markers in the peripheral blood which can detected either by ELISA or flow cytometry and give us insight into neuroinflammation or microglial activation? This is for clinical samples.
I am looking for a procedure to damage the bacterial wall and therefore promote the release of lipopolysaccharides in the culture medium, without killing the bacteria. I need to do this for develop a positive control of neuroinflammation by gut microbiota. Could you please suggest me also a E. coli strain or other kind of bacteria useful for this goal?
I have been recently trying to isolate primary microglia from mice pups brain (P0-P1). To prepare mixed microglia culture, I used 3 brains for a 75 cm2 flask. After around 10-12 days, astrocyte confluent layer is formed, and many microglia appears. I isolated those microglia by a shaking method (100-120 rpm/ 2 hrs).
At first harvest, I usually get 3-5 x 105 cells per flask. But, after the first harvest, I do not see the growth of new microglia. I only see the microglia that did not detach during the first harvest. Nevertheless, I tried to isolate for second harvest (after 1-2 weeks from 1st harvest), but end up with very low yield (2 x 104 cells).
Is there any ways to increase the yield?
Medium used: DMEM (10% FBS, 1% penicillin/streptomycin)
I am using SHSY5Y cells to develop neuroinflammation. I wanted to use MAP2 as a marker, but MAP2 only indicates the cells are differentiated. Is it necessary to ensure the cells are mature (eg: have synapse (synaptophysin as marker) to validate my in vitro model of neuroinflammation?
i am trying to characterise my LPS model (rat) with qPCR analysis to check the level of neuroinflammation, but i have a pretty big variance between the LPS treated animals.
excluding the differences between animals and how the may naturally deal with inflammation, do you think that specifically LPS may act so differently into animals and induce so different responses?
I rearching effect of estrojen on neuroinflammation with TNF-a. I will use JAK-STAT singnaling pathway and I will control differantiated RA Tnf(-), tnf(+) and tnf(+)östrojen(+). I will look at Nf- kappa B gene expression but I need to explain it detalied. How can I find detailed?
I've been looking for a suitable model of chronic neuroinflammation, that not involves a Neurodegenerative disease directly. So, I look for LPS model and TBI, wondering which model is better for evaluate chronic microglial activation.
Hello, we are analyzing inflammatory markers (such as TNF-alpha, IL1B, IL6 and INF-gamma) in brain samples (hippocampus and prefrontal cortex) after an intra-peritoneal injection of LPS in C57BL/6 mice.
Despite the high increments of these markers described in the literature using this method, we obtain very low increments or no changes. We suspect that maybe the fact that our animals are housed in a SPF facility could be interfering. Has anyone using SPF animals had the same problem? Could you give us some advice?
Currently, I am studying about neuroinflammation using murine derived microglial cell line. We focuse to reduce the inflammatory cytokines and inflammatory mediators production. In the near future we also hope to learn about amyloid-β downregulation. Is there any one who could give a suggestion regarding the mechanism linking between neuroinflammation and amyloid-β production ?
I am performing a study in which adrenalectomized (ADX) animals are going to be introduced to a chronic stress protocol and then having its corticosterone (CORT) levels analyzed a few days after the stress protocol. However, there is a growing field of literature reporting that ADX already induces severe neurodegeneration and neuroinflammation in these animals after 1-2 weeks (which are fewer days than what I intend to do). Thus, adding exogenous CORT would be an important control in such case.
I would like to have advice or previous experiences of researchers that did such work and their methods of controlling for CORT in this case. Thank you very much.
I am searching for the base of diseases with unkown course. I want to understand the base of low grade inflammation and the molecules involved in it.
I am looking for a way to use fluorescent microscopy in order to measure the levels of hydrogen peroxide in C. Elegans which exhibit Alzheimer’s symptoms like neurodegeneration. It is ok if there are two separate strains which specifically have some sort of marker for either hydrogen peroxide, catalase, or oxidative stress. Also, if you mention any strains, please link the article from which it is from.
Thanks in advance
Please I need your experience in understanding the role of Autophagy in Alzheimer's disease and if it could be a potential target for finding a cure for AD.
I am an MRI physicist, who is trying to understand the role of glutamate in neuro-inflammation. Since I have a limited understanding of biology/biochemistry, I would appreciate if someone can help me understand the role of glutamate in the inflammation.
I found the following overview very informative: (http://neurotransporter.org/glutamate.html ); however, it still fails to explain my basic question:
i) Does excessive extra-cellular glutamate always means that ONLY inflammation related processes are contributing? Could there be other contributing processes?
I would appreciate if someone can help me understand this.
I plan to study about the mechanism of anti neuro inflammation using cell culture method. My senior has conducted a similar experiment using mouse microglia MG6 cell line, but I want to uderstand the mechanism if the culture is changed by BV2 cell line, is there any signifficant differentiation between those cell culture type ?
Among the drugs used for MS, few like IFN-beta, Natalizumab, Glatiramer acetate don't cross BBB while Fingolimod do. So a humble request to the experts for their comments.
Has anyone used the tubes with gradient centrifugation to help separate out PMBCs ? Was the yeild as good or better than using the longer spin and no brake. Is it really that simple, you poor off the top later? Alll seems too good to be try - but expensive. I have some samples to try but would appreciate some prior knowledge
At what condition do I have to treat cells with LPS? In serum free media? For how many hours?
When he gives extract orally for 14 days and challenged by lps once, and performed novel object recognition. When he analysed his data, results was not dose dependent 100 and 200 mg is dose dependent but they declined in 400 mg but toxicity studies says (literature) LD may be up to 2000 mg. So what are all the possible reasons for this case.
Please share your suggestions
Thanks in advance
I injected a drug (subcutaneous in mice) and I noticed that there was an increase in number of microgias in the hippocampus (IBA positive cells). These microglias are more branched and non-reactive to MHC-II.
How should I interpret this increased in the number of branches and increased in the number of not activated microglia caused by a drug compared to control animals (which did not suffer intervention)? It can be interpreted as neuroprotection phenotype even if they have not suffered any inflammatory stimulus? Is it possible that microglia become more "resting" than those of the control animals that do not have inflammatory stimulus?
I've been culturing mixed glia and getting microglia from them for years, but it's always struck me that they look slightly yellow under the microscope. If you take a lysate of microglial cell as well, if you look at it, it is yellow-tinged too! Does anybody know what it is that the microglia are full of that gives them this colour? I've never seen this with any other cell type. Thanks!
I am planning to do some experiments in which I need to determine the levels of neuroinflammatory markers such as IL-6, TNF-alpha and IL-1beta in mice brain.
Hello everyone! I would like to find a lab host to participate to TALENTs grant. It is a ìn european grant that provide salary for 18 months, divided in 12 to spent in an Host institution and 6 to return to my base institution. I'm looking for an host laboratory situated in one of following country: Slovenia*, Croatia, Bosnia and Herzegovina, Serbia, Montenegro, Albania, Greece, Germany, France, Austria, Switzerland, Liechtenstein. The laboratory, given my CV, should ideally work in neuroinflammation / neurodegenerative diseases or I would like to learn CRISPR/CAS9 system...details in the link attached..
thanks for all the possible answers!
To improve data and reduce variability i would like to move away from outbred strain (Sprague Dawley) and use inbred rats. But it is nearly impossibly to work out which strain to use from the available ones: Lewis, Long Evans, Fischer, Kyoto and many more. I know, for example, that Lewis rats are used for some neuroimmunology (EAE model) but this is different to our model (effect of systemic LPS on the brain) and therefore might not be directly relevant. Do people have any experience with this type of work and inbred rats?
We are going to do improvement of cancer chemotherapy drugs-induced cognitive impairment and peripheral neuropathy via inhibition of neuroinflammation and oxidative stress by using natural compound. We will use paclitaxel as a chemo drug. We are searching for doses of paclitaxel in rat which is related to human.Thank you.
Macrophages are known to be an active player in MS. They are involved in active demyelination and myeline uptake, which leads towards the development of foamy macrophages with M2-like properties. But can they migrate out of the brain/lesion before they become necrotic and cause a pro-inflammatory respons (cfr. atherosclerosis)?
I'd like to demonstrate that some medicine could reduce the neuroinflammation in ICH model via in vitro and in vivo study. Could raw 264.7 macrophages be used in vitro study instead of BV2 microglia (because microglia was infected by mycoplasma)? What are the differences between them ?
I am considering using a scholl analysis (using imagej software) but I was wondering if anyone has any better ideas. I only need to determine if the microglia are activated, I don't need to describe their morphology in detail.
If you are aware of the fact that cancer patients treated with standard chemotherapy over the past 20 or so years have cognitive problems with reasoning and motor skills, I'd like your input on why, and why ten to twenty years later. Microglia (brain macrophages, which is a misnomer) can initiate the process of neurodegeneration, which leads to ex-cancer patients, who've survived through chemotherapy, down the path of significant brain damage. There is an initial insult due to systemic inflammation, but I'm thinking that endothealial damage and their ability to secrete inflammatory lymphokines perpetuate or exhasterbate chemotherapy-induced cognitive problems in cancer patients.
What do you think?
Concerning SPIO and macrophages in neuroinflammation.
What is the safe dose and how to assess the toxic effect of SPIO on macrophages in vitro?
I am researching into in vitro models linked to inflammation and depression, not ex-vivo or from primary microglia.
Any help would be appreciated.
I am looking for a protocol to quantify proteins in the mice brain by western blot technique. I want to see the effect of treatment on AD pathology and neuroinflammation markers. I would like to analyze following proteins: Tau, BACE1, COX-2, GFAP, IL-6, IL-1beta, TNF-alpha, iNOS and p38 MAK. Can somebody share with me the wetern blot protocol with all necessary details. Thank you in advance.
Does somebody have experience in inducing the microglial activation by systemic LPS injection in rats? Mice seem to respond well to LPS, but rat do not demonstrate neuroinflammatory changes in response to i.p. LPS injection. The data in PubMed concerning this issue are very sparse, but there is still a possibility that some negative results remained unpublished.
First it was the death of dopaminergic neurons as a result of "oxidative stress". Then the misfolding of α-synuclein. Now there si a suggestion
(http://www.foxnews.com/health/2012/05/16/early-signs-parkinson-might-be-seen-in-colon/#) that α-synuclein is found in the colon in in PD.
I had a 2 week course of prednsiolone (30mg.day) last September for an unrelated condition and am now on an 8 week course of budesonide (9 mg/day) these abolish all joint stiffness and pain and I can now climb stairs easily, turn over in bed without having to plan it and gather strength, leap out of chairs and look over my shoulder. I still walk slowly and still fall over more than I used to and my speech is no better but I'm not mildly incontinent any more!!
I was for a week taking 5-amino salicylic acid and this was, I think, just as effective.
It looks as if some PD symptoms are caused by inflammation and may be easily and safely treatable. I found references to inflammation in PD:
but I cannot see any clear connection to α-synuclein.
Is there a coherent story somewhere or are we dealing with more than one related but separate conditions with different causes?
I am facing problems while counting Iba 1 positive cells in mouse brain section. I cut the brain slices in 20 micron thickness and performed immunohistochemistry. And now I have to perform the quantification of Iba 1 positive cells in the sections. A senior researcher just suggested to count all cells present in the photographs of the both cortex and striatum. But I think this is not the appropriate way of counting. For your convenience, I have attached a photograph here. Could you please let me know how many Iba1 positive cells are present there? Remember the thickness of the slice is 20 micrometer.
Is it possible that rats and mice differ in their susceptibility to LPS (E.coli) effects? We have mentioned serious microglial activation in the brain (increase in the number and cell area of microglial cells) after intraperitoneal LPS injection in mice (2 mg/kg), but rats (Wistar) seem insensitive to LPS under the same conditions (I mean, in the CNS - no activation of microglia measured by cell area and cell number). It was rather surprising to us since we did not suspect such an inter-species difference in the course of inflammatory response. Do you have any assumptions about why it could happen, given that all experimental conditions were similar for rats and mice? Has anyone noted the difference in the timecourse and/or intensity of inflammatory response between these species? I would be grateful for any ideas.
Does neuroinflammation and neurodegeneration induced by continuous infusion (intracerebroventricular) of LPS occur in all CNS neurons or only in specific regions? The literature is rich in research about the hippocampus, but not on other regions. I have special interest in the hypothalamus.
TNF alpha concentration, its duration, and the activated mechanism can produce opposite effects, both protective or noxious. (Montgomery 2012)
Tumor necrosis factor alpha (TNFa) promotes synaptic scaling and gliotransmission at glutamatergic synapses via complementary post and presynaptic actions (Santello 2012).
increasing AMPA receptors lead to more excitation of post synaptic neuron and result of that is exit more K+ from neuron which maybe have effects on astrocyte. But what effects?
A-804598 is an antagonist for P2X7 receptors. We're trying to find out if there's an in vivo data in relation to toxicity for this particular antagonist. Please let me know if there's anything, much appreciated.
- It's an 8 weeks chronic mild stress mice model, interest being the polarization of microglia/macrophages and NLRP3 inflammasome activation in CNS. Administration route is via gavaging syringe. (updated 08/04/2014)
I am attempting to measure brain inflammation in lysates from cortex tissue. Is there any test that is a good "overall" measure of brain inflammation from these lysates?