Cell coupling and Cx43 expression in embryonic mouse neural progenitor cells

Embryonic neural progenitors isolated from the mouse striatal germinal zone grow in vitro as floating cell aggregates called neurospheres, which, upon adhesion, can be induced to differentiate into the three main cell types of the central nervous system (CNS), that is, astrocytes, neurons and oligodendrocytes. To study the possible role of connexins and junctional communication during differentiation of neural progenitors, we assessed cell-to-cell communication by microinjecting Lucifer Yellow into neurospheres at various times after adhesion. Cells located in neurospheres were strongly coupled, regardless of the differentiation time. Microinjections performed on the cell layers formed by differentiated cells migrating out of the neurosphere established that only astrocytes were coupled. These observations suggest the existence of at least three distinct communication compartments:coupled proliferating cells located in the sphere, uncoupled cells undergoing neuronal or oligodendrocytic differentiation and coupled differentiating astrocytes. A blockade of junctional communication by 18-β-glycyrrhetinic acid (βGA) reduced, in a concentration-dependent manner, the viability of undifferentiated neural progenitor cells. This effect appeared to be specific,inasmuch as it was reversible and that cell survival was not affected in the presence of the inactive analog glycyrrhyzic acid. Addition of βGA to adherent neurospheres also decreased cell density and altered the morphology of differentiated cells. Cx43 was strongly expressed in either undifferentiated or differentiated neurospheres, where it was found both within the sphere and in astrocytes, the two cell populations that were dye coupled. Western blot analysis further showed that Cx43 phosphorylation was strongly increased in adherent neurospheres, suggesting a post-translational regulation during differentiation. These results point to a major role of cell-to-cell communication and Cx43 during the differentiation of neural progenitor cells in vitro.

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Temporal and epigenetic regulation of neurodevelopmental plasticity

Philosophical Transactions of the Royal Society B Biological Sciences ◽

10.1098/rstb.2006.2010 ◽

2007 ◽

Vol 363 (1489) ◽

pp. 23-38 ◽

Cited By ~ 21

Author(s):

Nicholas D Allen

Keyword(s):

Stem Cells ◽

Progenitor Cells ◽

Developmental Plasticity ◽

Neural Progenitor Cells ◽

Embryonic Stem ◽

Neural Progenitor ◽

Neural Progenitors ◽

Developmental Potential ◽

Neural Lineage

The anticipated therapeutic uses of neural stem cells depend on their ability to retain a certain level of developmental plasticity. In particular, cells must respond to developmental manipulations designed to specify precise neural fates. Studies in vivo and in vitro have shown that the developmental potential of neural progenitor cells changes and becomes progressively restricted with time. For in vitro cultured neural progenitors, it is those derived from embryonic stem cells that exhibit the greatest developmental potential. It is clear that both extrinsic and intrinsic mechanisms determine the developmental potential of neural progenitors and that epigenetic, or chromatin structural, changes regulate and coordinate hierarchical changes in fate-determining gene expression. Here, we review the temporal changes in developmental plasticity of neural progenitor cells and discuss the epigenetic mechanisms that underpin these changes. We propose that understanding the processes of epigenetic programming within the neural lineage is likely to lead to the development of more rationale strategies for cell reprogramming that may be used to expand the developmental potential of otherwise restricted progenitor populations.

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Corrections to “Light Supports Neurite Outgrowth of Human Neural Progenitor Cells In Vitro: The Role of P2Y Receptors” [Jan/Feb 08 118-125]

IEEE Journal of Selected Topics in Quantum Electronics ◽

10.1109/jstqe.2008.921188 ◽

2008 ◽

Vol 14 (2) ◽

pp. 521-521 ◽

Cited By ~ 1

Author(s):

J. J. Anders ◽

T. B. Romanczyk ◽

I. K. Ilev ◽

H. Moges ◽

L. Longo ◽

...

Keyword(s):

Neurite Outgrowth ◽

Progenitor Cells ◽

Neural Progenitor Cells ◽

P2y Receptors ◽

Neural Progenitor ◽

Human Neural Progenitor Cells

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Wnt Receptor Frizzled-4 as a Marker for Isolation of Enteric Neural Progenitors in Human Children

Cells ◽

10.3390/cells8080792 ◽

2019 ◽

Vol 8 (8) ◽

pp. 792

Author(s):

Peter H. Neckel ◽

Melanie Scharr ◽

Karin Seid ◽

Katharina Nothelfer ◽

Jörg Fuchs ◽

...

Keyword(s):

Nervous System ◽

Progenitor Cells ◽

Cell Sorting ◽

Neural Progenitor Cells ◽

Human Colon ◽

Neural Progenitor ◽

Neural Progenitors ◽

Proliferation Assays ◽

Sodium And Potassium

Identification and isolation of neural progenitor cells from the human enteric nervous system (ENS) is currently hampered by the lack of reliable, specific markers. Here, we define the Wnt-receptor frizzled-4 as a marker for the isolation of enteric neural progenitor cells derived from paediatric gut samples. We show that the Wnt-receptor frizzled-4 is expressed in the human colon and in Tunica muscularis-derived enterospheres. To obtain a purified culture, we carried out fluorescence-activated cell sorting (FACS) using PE-conjugated frizzled-4 antibodies. Frizzled-4positive cells gave rise to neurosphere-like bodies and ultimately differentiated into neurons as revealed by BrdU-proliferation assays and immunocytochemistry, whereas in frizzled-4negative cultures we did not detect any neuronal and glial cells. By using a patch-clamp approach, we also demonstrated the expression of functional sodium and potassium channels in frizzled-4positive cell cultures after differentiation in vitro.

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Myelin basic protein enhances axonal regeneration from neural progenitor cells

Cell & Bioscience ◽

10.1186/s13578-021-00584-7 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Zhengjian Yan ◽

Lei Chu ◽

Xiaojiong Jia ◽

Lu Lin ◽

Si Cheng

Keyword(s):

Myelin Basic Protein ◽

Neurite Outgrowth ◽

Progenitor Cells ◽

Axonal Regeneration ◽

Neural Progenitor Cells ◽

Basic Protein ◽

Neural Progenitor ◽

Dependent Manner ◽

Cord Injury

Abstract Introduction Stem cell therapy using neural progenitor cells (NPCs) shows promise in mitigating the debilitating effects of spinal cord injury (SCI). Notably, myelin stimulates axonal regeneration from mammalian NPCs. This led us to hypothesize that myelin-associated proteins may contribute to axonal regeneration from NPCs. Methods We conducted an R-based bioinformatics analysis to identify key gene(s) that may participate in myelin-associated axonal regeneration from murine NPCs, which identified the serine protease myelin basic protein (Mbp). We employed E12 murine NPCs, E14 rat NPCs, and human iPSC-derived Day 1 NPCs (D1 hNPCs) with or without CRISPR/Cas9-mediated Mbp knockout in combination with rescue L1-70 overexpression, constitutively-active VP16-PPARγ2, or the PPARγ agonist ciglitazone. A murine dorsal column crush model of SCI utilizing porous collagen-based scaffolding (PCS)-seeded murine NPCs with or without stable Mbp overexpression was used to assess locomotive recovery and axonal regeneration in vivo. Results Myelin promotes axonal outgrowth from NPCs in an Mbp-dependent manner and that Mbp’s stimulatory effects on NPC neurite outgrowth are mediated by Mbp’s production of L1-70. Furthermore, we determined that Mbp/L1-70’s stimulatory effects on NPC neurite outgrowth are mediated by PPARγ-based repression of neuron differentiation-associated gene expression and PPARγ-based Erk1/2 activation. In vivo, PCS-seeded murine NPCs stably overexpressing Mbp significantly enhanced locomotive recovery and axonal regeneration in post-SCI mice. Conclusions We discovered that Mbp supports axonal regeneration from mammalian NPCs through the novel Mbp/L1cam/Pparγ signaling pathway. This study suggests that bioengineered, NPC-based interventions can promote axonal regeneration and functional recovery post-SCI.

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