title: Light-controlled astrocytes promote human mesenchymal stem cells toward neuronal differentiation and improve the neurological deficit in stroke rats.
abstract: Astrocytes are key components of the central nervous system (CNS) and release factors to support neural stem cell proliferation, differentiation, and migration. Adenosine 5'-triphosphate (ATP) is one of the key factors released upon activation of astrocytes that regulates the neural stem cell's function. However, it is not clear whether ATP derived from the depolarized astrocytes plays a vital role in promoting the neuronal differentiation of mesenchymal stem cells (MSCs) in vitro and in vivo. Herein, for the first time, we co-cultured MSCs with light-stimulated-channelrhodopsin-2 (ChR2)-astrocytes, and observed that the neuronal differentiation of MSCs was enhanced by expressing more neuronal markers, Tuj1 and NeuN. The ChR2-astrocyte-conditioned medium also stimulated MSCs differentiating into neuronal lineage cells by expressing more Tuj1 and Pax6, which was blocked by the P2X receptor antagonist, TNP-ATP. Then we found that light-depolarization of astrocytes significantly increased ATP accumulation in their bathing medium without impairing the cell membrane. We further found that ATP up-regulated the Tuj1, Pax6, FZD8 and β-catenin mRNA levels of MSCs, which could be reversed by application of TNP-ATP. Together these in vitro data provided convergent evidence that ATP from light-depolarized-astrocytes activated the wnt/β-catenin signaling of MSCs through binding to the P2X receptors, and promoted the neuronal differentiation of MSCs. Finally but importantly, our study also demonstrated in stroke rats that light-controlled astrocytes stimulated endogenous ATP release into the ischemic area to influence the transplanted MSCs, resulting in promoting the MSCs towards neuronal differentiation and improvements of neurological deficit.
authors.affiliation: Shenzhen Key Lab of Neuropsychiatric Modulation, Key Laboratory for Health Informatics at Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, People's Republic of China.
authors.forename: Jie
authors.initials: J
authors.lastname: Tu
authors.forename: Fan
authors.initials: F
authors.lastname: Yang
authors.forename: Jun
authors.initials: J
authors.lastname: Wan
authors.forename: Yunhui
authors.initials: Y
authors.lastname: Liu
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.forename: Bifeng
authors.initials: B
authors.lastname: Wu
authors.forename: Yafeng
authors.initials: Y
authors.lastname: Liu
authors.forename: Shaoqun
authors.initials: S
authors.lastname: Zeng
authors.forename: Liping
authors.initials: L
authors.lastname: Wang
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000502:physiology*; D002454:Cell Differentiation; D049109:Cell Proliferation; D064987:Cell- and Tissue-Based Therapy; D002478:Cells, Cultured; D000075402:Channelrhodopsins; D003342:Corpus Striatum / Q000166:cytology; D004195:Disease Models, Animal; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000502:physiology; D009422:Nervous System Diseases / Q000209:etiology* / Q000628:therapy*; D051381:Rats; D017207:Rats, Sprague-Dawley; D020521:Stroke / Q000150:complications* / Q000628:therapy*; D014161:Transduction, Genetic
chemical_list: D000075402:Channelrhodopsins
keywords: ATP; astrocytes; channelrhodopsin-2 (ChR2); mesenchymal stem cells (MSCs); microdialysis; stroke