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In Vitro Cellular & Developmental Biology - Animal 02/2013; · 1.31 Impact Factor
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Jinhong Shi,
Haoming Li, Guohua Jin,
Peipei Zhu,
Meiling Tian,
Jianbing Qin,
Xuefeng Tan,
Shuqing Zhao,
Fuyu Wang,
Yurong Hua,
Yong Xiao
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ABSTRACT: Lhx8, also named L3, is a recently identified member of the LIM homeobox gene family. Previously, we found acetylcholinesterase (AChE)-positive cells in fimbria-fornix (FF) transected rat hippocampal subgranular zone (SGZ). In the present study, we detected choline acetyltransferase (ChAT)-positive cholinergic cells in hippocampal SGZ after FF transaction, and these ChAT-positive cells were double labeled by Lhx8. Then we overexpressed Lhx8 during neural differentiation of hippocampal neural stem/progenitor cells on adherent conditions using lentivirus Lenti6.3-Lhx8. The result indicated that overexpression of Lhx8 did not affect the proportion of MAP2-positive neurons, but increased the proportion of ChAT-positive cells in vitro. These results suggested that FF-transected hippocampal niche promoted the ChAT/Lhx8-positive cholinergic neurons generation in rodent hippocampus, and Lhx8 was not associated with the MAP2-positive neurons differentiation on adherent conditions, but played a role in the specification of cholinergic neurons derived from hippocampal neural stem/progenitor cells in vitro.
In Vitro Cellular & Developmental Biology - Animal 11/2012; · 1.31 Impact Factor
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ABSTRACT: Neural stem cells (NSCs) tranplantation has great potential for the treatment of neurodegenerative disease such as Parkinson's disease (PD). However, the usage of NSCs is limited because the differentiation of NSCs into specific dopaminergic neurons has proven difficult. We have recently demonstrated that transgenic expression of Nurr1 could induce the differentiation of NSCs into tyrosine hydroxylase (TH) immunoreactive dopaminergic neurons, and forced co-expression of Nurr1 with Brn4 caused a dramatic increase in morphological and phenotypical maturity of these neurons. In this study, we investigated the effect of transplanted NSCs in PD model rats. The results showed that overexpression of Nurr1 promoted NSCs to differentiate into dopaminergic neurons in vivo, increased the level of dopamine (DA) neurotransmitter in the striatum, resulting in behavioral improvement of PD rats. Importantly, co-expression of Nurr1 and Brn4 in NSCs significantly increased the maturity and viability of dopaminergic neurons, further raised the DA amount in the striatum and reversed the behavioral deficit of the PD rats. Our findings provide a new potential and strategy for the use of NSCs in cell replacement therapy for PD.
International journal of developmental neuroscience: the official journal of the International Society for Developmental Neuroscience 10/2012; · 2.03 Impact Factor
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ABSTRACT: We isolated and expanded fibroblast-like cells from the Wharton's jelly of human umbilical cord successfully. Immunocytochemistry showed that they were positive for several markers of mesenchymal stem cells (CD73, CD90, and CD105) and integrin markers (CD29 and CD44), but negative for a hematopoietic cell maker (CD45) and an endothelial cell marker (CD31). Their differentiation into osteocytes and adipocytes under specific conditions indicated that they had multi-lineage differentiation potential. Therefore these results proved that the cells we obtained from Wharton's jelly were human umbilical cord mensenchymal stem cells (hUCMSCs). Using immunocytochemistry and Western blotting analysis, we found that after treatment with neuronal induction medium [NIM; consisting of brain-derived neurotrophic factor (BDNF) and low-serum media] for 14 days, hUCMSCs expressed a neuronal specific marker, microtubule associated protein 2 (MAP2), and extended neurite-like processes. After treatment with NIM, supplemented with hippocampal cholinergic neurostimulating peptide (HCNP) or rat denervated hippocampal extract [rDHE; derived from rat fimbria fornix (FF) transected hippocampus], hUCMSCs expressed choline acetytransferase (ChAT) and this action could be enhanced when cells were cultured with NIM, supplemented with HCNP and rDHE in combination. ELISA showed that these ChAT-positive cells could secrete acetylcholine (ACh). These findings indicate that hUCMSCs possess the potential of differentiation into functional ChAT-positive cells in vitro and provide a new candidate of cells for the cell transplantation to treat Alzheimer's disease (AD).
International journal of developmental neuroscience: the official journal of the International Society for Developmental Neuroscience 06/2012; 30(6):471-7. · 2.03 Impact Factor
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ABSTRACT: Mash1, a member of the basic helix-loop-helix (bHLH) transcription factor family, has previously been considered essential for neuronal differentiation and specification in the nervous system. In this study, we investigated the expression of Mash1 in the hippocampus after fimbria-fornix (FF) transection. Western blot showed that protein of Mash1 increased significantly and peaked at day 7 after FF transection. Immunofluorescence indicated that after FF transection, more newborn cells differentiated into Mash1 positive cells in the deafferented side than that in the normal side, and we investigated that in the neurogenic area, subgranular zone (SGZ), a part of Mash1 positive cells were NeuN positive, and more Mash1/NeuN double positive neurons were identified in the deafferented side than that in the normal side. Additionally, the number of Mash1/NeuN double positive neurons in SGZ increased significantly and peaked at day 7 after FF transection. In vitro, immunofluorescence revealed that extracts of the deafferented hippocampus promoted neuronal differentiation to a greater extent than extracts from normal hippocampus. Deafferented extracts also enhanced Mash1 expression in MAP-2 positive neurons. This study concludes that after FF transection, Mash1 expression in the deafferented hippocampus increased and might play an important role in inducing local progenitors to differentiate into neurons.
Neuroscience Letters 05/2012; 520(1):26-31. · 2.11 Impact Factor
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02/2012; , ISBN: 978-953-307-958-5
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ABSTRACT: During the central nervous system (CNS) development, radial glia cells (RGCs) play at least two essential roles, they contribute to neuronal production and the subsequent guidance of neuronal migration, whereas its precise distribution and contribution to cerebral cortex remains less understood. In this research, we used Vimentin as an astroglial marker and Sox2 as a neural progenitor marker to identify and investigate RGCs in rat cerebral cortex at embryonic day (E) 16.5. We found that the Sox2+ progenitor cells localized in the germinal zone (GZ) of E16.5 cerebral cortex, ~95% Sox2+ cells co-localized with Vimentin+ or Nestin+ radial processes which extended to the pial surface across the cortical plate (CP). In vitro, we obtained RG-like cells from E16.5 cerebral cortex on adherent conditions, these Sox2+ Radial glia (RG)-like cells shared some properties with RGCs in vivo, and these Sox2+ RG-like cells could differentiate into astrocytes, oligodendrocytes and presented the radial glia-neuron lineage differentiation ability. Taken together, we identified and investigated some characterizations and properties of Sox2+ RGCs derived from E16.5 cerebral cortex, we suggested that the embryonic Sox2+ progenitor cells which located in the cortical GZ were mainly composed of Sox2+ RGCs, and the cortex-derived Sox2+ RG-like cells displayed the radial glia-neuron lineage differentiation ability as neuronal progenitors in vitro.
Histochemie 09/2011; 136(5):515-26. · 2.59 Impact Factor
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ABSTRACT: Successful neural stem cells (NSCs) therapies require the controlled differentiation of NSCs into neurons. Porous chitosan scaffold was explored if it promoted neuronal differentiation of NSCs in the presence of nerve growth factor (NGF) in 3-dimensional (3-D) culture.
Chitosan scaffold was made by the freeze-drying technique. NSCs were cultured under four different conditions: on flat cover slips (2-D structure) in media with or without NGF, and on chitosan scaffold (3-D structure) in media with or without NGF. Immunohistochemical staining was used to observe multi-directional differentiation of cultured NSCs. Photomicrographs were taken and analyzed for cell number, soma size, and neuronal process length.
The porosity index of chitosan scaffold was around 90% and the diameter of pores was 50-350 μm. NSCs could differentiate into neurons, astrocytes, and oligodendrocytes under all culture conditions. The rank efficacy for neuronal differentiation was 3-D culture with NGF group > 3-D culture without NGF group > 2-D culture with NGF group > 2-D culture without NGF group.
The results suggest that the combination of chitosan scaffold and NGF exerts a synergistic effect on neuronal differentiation of NSCs, a requirement for successful integration into the damaged central nervous system.
Neuro endocrinology letters 09/2011; 32(5):705-10. · 1.30 Impact Factor
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ABSTRACT: To explore the effects of deafferented hippocampal extracts on the differentiation of radial glial cells (RGCs), hippocampal RGCs of postnatal day 1 rats were isolated under adherent conditions in vitro. Protein extracts of deafferented hippocampus were prepared from adult rats following fimbria fornix lesion. RGCs were exposed to extracts of deafferented or normal hippocampus and the type and extent of proliferation and differentiation were evaluated. We report that extracts of deafferented hippocampus more effectively promoted RGC proliferation than extracts of normal hippocampus. Moreover, although RGC differentiation in vitro primarily generated cells of glial lineages, cells exposed to extracts of deafferented hippocampus, but not of normal hippocampus, showed a significantly increased trend towards the generation of cells of neuronal lineages. We conclude that extracts of deafferented hippocampus promote RGC proliferation and neurogenesis.
Neuroscience Letters 07/2011; 498(1):93-8. · 2.11 Impact Factor
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ABSTRACT: Fetal brain tissue can be used in cell replacement therapy for PD (Parkinson's disease), but there is a poor donor supply of this tissue. NSCs (neural stem cells) may overcome this problem as they can be isolated and expanded in vitro. However, the usage of NSCs is limited because the differentiation of NSCs into specific dopaminergic neurons has proven difficult. In the present study, we investigated the effect of Nurr1 (nuclear receptor related factor 1), a transcription factor specific for the development and maintenance of the midbrain dopaminergic neurons on inducing the differentiation of NSCs into TH (tyrosine hydroxylase) immunoreactive dopaminergic neurons. Nonetheless, these cells exhibited an immature neuronal morphology with small cell bodies and short neurite processes, and they seldom expressed DAT (dopamine transporter), a late marker of mature dopaminergic neurons. However, forced co-expression of Nurr1 with Brn4, a member of the POU domain family of transcription factors, caused immature Nurr1-induced dopaminergic neurons to differentiate into morphologically and phenotypically more mature neurons. Thus the enriched generation of mature dopaminergic neurons by forced expression of Nurr1 with Brn4 may be of future importance in NSC-based cell replacement therapy for PD.
Cell Biology International 06/2011; 35(12):1217-23. · 1.48 Impact Factor
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ABSTRACT: The role of radial glia cells (RGCs) as neural progenitors and as guides for migrating neurons is well established, mouse or human-derived radial glia (RG)-like cells in vitro showed some astroglia and stem/progenitor properties like RGCs in vivo, but different species-derived RG-like cells present some different properties. Here we acquired rat-derived RG-like cells on adherent conditions in vitro and then identified their astroglia and stem/progenitor properties. Similarly to the RGCs, the RG-like cells could be double-labeled by brain lipid-binding protein, glial fibrillary acidic protein, vimentin with nestin and expressed some astroglia and stem/progenitor genes; these cells also presented tripotent differentiation potentialities, albeit the ability of gliogenesis far exceeded the neurogenesis in vitro. Taken together, we acquired and identified some properties of rat-derived RG-like cells from fetal cerebral cortices in vitro.
In Vitro Cellular & Developmental Biology - Animal 05/2011; 47(7):431-7. · 1.31 Impact Factor
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ABSTRACT: The role of radial glia cells (RGCs) as neural progenitors and as guides for migrating neurons is well established, whereas their precise contribution to adult hippocampal neurogenesis remains less understood. To precisely study the properties of hippocampal RGCs under normal conditions in vitro, here we acquired the hippocampal RGCs of postnatal 1 d rats under adherent conditions in vitro, identified their astroglia and stem/progenitor properties. We found that the neonatal rat hippocampal RGCs had longer processes than the RGCs from fetal cerebral cortices, and these cells could be double-labeled by BLBP, GFAP, Vimentin with Nestin and expressed some stem/progenitor genes, these cells also presented multiple differentiation potentialities, albeit the ability of gliogenesis far exceeded the neurogenesis under normal culture conditions in vitro. Taken together, we acquired and identified some properties of the RGCs from neonatal rat hippocampi in vitro.
Neuroscience Letters 03/2011; 490(3):209-14. · 2.11 Impact Factor
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ABSTRACT: Extracts of deafferented hippocampus were previously found to promote neuronal differentiation of neural stem cells (NSCs). To gain insights into the underlying molecular mechanisms we studied the potential involvement of signal transducer and activator of transcription3 (STAT3) activation in the NSCs response to hippocampal extracts. Here we report that phosphorylated STAT3 (p-STAT3) is expressed at different stages in neurons and astrocytes differentiated from rat hippocampus-derived NSCs. Deafferented hippocampal extracts produced sustained upregulation of p-STAT3 levels and promoted NSC differentiation and neurogenesis, whereas extracts of normal hippocampus were without effect. Interleukin-6 (IL-6), an activator of JAK/STAT signaling pathways, had no effect on neurogenesis, whereas the selective STAT3 inhibitor p-ip-STAT3 decreased the number of Microtubule-associated protein-2 (MAP-2)-positive cells generated by NSC differentiation. These findings argue that STAT3-related signaling pathways are likely to play a role in neuronal survival and differentiation during NSC neurogenesis stimulated by extracts of deafferented hippocampus.
Neuroscience Letters 02/2011; 493(1-2):18-23. · 2.11 Impact Factor
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ABSTRACT: Lesions to the fimbria fornix (FiFx) plus cingulate bundle (CB), the principal routes of communication of forebrain cholinergic regions, produce lasting impairment of spatial learning and memory in mice. We report that extensive neurogenesis takes place in the FiFx, CB, and basalis magnocellularis following FiFx plus CB transection. Immunofluorescence revealed that nestin-expressing cells were present in all 3 areas following lesion; the majority of nestin-positive cells were also positive for 5-bromo-2-deoxy-uridine, a marker of DNA synthesis. Nestin-positive proliferative cells were almost entirely absent from unlesioned tissue. Neurospheres cultured in vitro from lesioned FiFx displayed the characteristics of neural stem cells--proliferation, expression of embryonic markers, and multipotential differentiation into neurons, astrocytes, and oligodendrocytes. At early stages after transection, a small number of immature and migrating doublecortin-immunopositive neurons were detected in lesioned FiFx, where neuronal cell bodies are normally absent. At later stages, postlesion immature neurons developed into β-tubulin III-positive mature neurons. Lentivirus labeling assay implied that the injury-induced neurogenesis in FiFx may be from local neurogenic astrocytes but not from dentate gyrus. These results demonstrate that insult to cholinergic tracts can stimulate neural stem cell proliferation and neuronal regeneration not only in innervated regions but also in the projection pathways themselves. Ectopic neurogenesis in cholinergic system-related areas provides an additional mechanism for repair of cholinergic innervation following damage.
Stem cells and development 12/2010; 20(9):1627-38. · 4.15 Impact Factor
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ABSTRACT: Brn-4, a member of the homeobox family of transcription factors, has previously been implicated in the regeneration and repair of denervated striatum. We investigated the effects of Brn-4 on the differentiation and development of neural stem cells (NSCs) from E16 rat hippocampus. Immunocytochemistry revealed that extracts of deafferented hippocampus promoted neuronal differentiation to a greater extent than extracts from normal hippocampus. Deafferented extracts also promoted maturation of newborn neurons as reflected in changes in cell areas and perimeters, and enhanced Brn-4 expression in MAP-2 positive neurons. Suppression or overexpression of Brn-4 in NSCs markedly decreased or increased neuronal differentiation and maturation of newborn neurons, respectively. These results suggest that Brn-4 expression is required both for neuronal differentiation of NSCs and maturation of newborn neurons, and that there may be some regulatory factors in deafferented hippocampus that can regulate Brn-4 expression in neuronal progenitors. Brn-4 is therefore a potential research target for the development of new therapeutics to promote brain repair.
Neuroscience Research 05/2010; 67(1):8-17. · 2.25 Impact Factor
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ABSTRACT: Neurogenesis in the hippocampus continues throughout adult life and can be regulated by the local microenvironment. To determine whether denervation stimulates neurogenesis in hippocampus, proliferation, migration, and differentiation of local neural stem cells (NSCs) in dentate gyrus was investigated after fimbria fornix transection. In the denervated hippocampus, NSCs proliferated markedly and migrated along the subgranular layer, and more newborn cells differentiated into neurons or astrocytes. After denervation, more newborn cells in the deafferented hippocampus expressed Brn-4 and differentiated into beta-Tubulin III positive neurons. It is concluded that the local NSCs in hippocampus may proliferate and migrate into granule cell layer, in which changes in the deafferented hippocampus provided a suitable microenvironment for hippocampal neurogenesis and the increased Brn-4 in denervated hippocampus may be involved in this process.
The International journal of neuroscience 03/2010; 120(3):192-200. · 0.86 Impact Factor
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ABSTRACT: Fimbria-fornix (FF), the septo-hippocampal pathway, was transected to model Alzheimer's disease (AD), which is characterized by loss of cholinergic afferent fibers in hippocampus. Various alternations may happen in the deafferented hippocampus. In this study, we determined the expression of Brn-4 in hippocampus after FF lesion. RT-PCR and Western blot showed that mRNA transcription and protein of Brn-4 increased significantly and reached to the peak at day 14 after FF lesion. Hybridization and immunohistochemistry indicated that Brn-4 signals in hippocampus and dentate gyrus (DG) of the deafferented side were significantly stronger than the normal side. More Brn-4 positive cells were identified in the DG of deafferented hippocampus. In the pyramidal and granular cells, Brn-4 positive cells were all NeuN positive neurons, whereas in the neurogenic area, subgranular zone (SGZ), only a part of Brn-4 positive cells were NeuN positive, and these Brn-4/NeuN double positive neurons in SGZ and hilus of DG increased significantly after the trauma induced by FF lesion. In vitro Brn-4 antibody attenuated the role of extract from deafferented hippocampus in promoting differentiation of hippocampal progenitors into MAP-2 positive neurons. This study demonstrated that after FF lesion, Brn-4 in the deafferented hippocampus was upregulated and might play an important role in inducing local progenitors to differentiate into neurons, which may compensate for the loss of cholinergic afferent fibers or other dysfunctions.
Hippocampus 11/2008; 19(2):176-86. · 5.18 Impact Factor
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ABSTRACT: The fate of neural stem/progenitor cells (NSCs/NPCs) in vivo lies on the local microenvironment. Whether the denervated hippocampus provides a stimulative role on the survival and differentiation of the anterior subventricular zone (SVZa) progenitors was investigated in the present study. In vivo the SVZa progenitors were transplanted into the denervated hippocampus and the contralateral side, and were found migrating along the subgranular layer. More implanted cells were found survived and differentiated into the Neurofilament 200 (NF-200) or beta-Tubulin-III positive neurons in the denervated than in the normal hippocampus at all points studied. In vitro the extracts from the denervated and normal hippocampus were used to induce differentiation of the SVZa progenitors. More progenitors incubated with the denervated hippocampal extract differentiated significantly into the MAP-2 or AChE positive neurons than those incubated with the normal hippocampal extract (P<0.05). We concluded that the deafferented hippocampus provided proper microenvironment for the survival and neuronal differentiation of neural progenitors.
Neuroscience Letters 04/2007; 414(2):115-20. · 2.11 Impact Factor
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ABSTRACT: The expansion of human neural stem cells in vitro might overcome the poor donor supply of human fetal neural tissue in transplantation for Parkinson's disease. However, the differentiation of human neural stem cells into dopaminergic neurons has proven difficult. In the present study, we investigated the effects of cytokines, trophic factors of developmental striatum and Ginkgolide on differentiation of human neural stem cells (hNSCs) into TH-ir neurons. The immunoreactivity to tyrosine hydroxylase (TH), a distinctive marker for dopamine neurons was used to assess dopaminergic neuronal phenotype. We demonstrate that human neural stem cells expanded in vitro can efficiently differentiate into TH-ir neurons by induction. These stem cells might serve as a continuous, on-demand source of cells for therapeutic transplantation in patients with Parkinson's disease.
Neuroscience Letters 10/2005; 386(2):105-10. · 2.11 Impact Factor
