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retinal regeneration requires dynamic Notch signaling

Neural regeneration Research 2022年第6期17卷 1199-1209页

作者： Leah J.Campbell Jaclyn L.Levendusky Shannon A.Steines David R.Hyde Department of Biological Sciences Center for Zebrafish ResearchCenter for Stem Cells and Regenerative MedicineUniversity of Notre DameNotre DameINUSA

retinal damage in the adult zebrafish induces Müller glia reprogramming to produce neuronal progenitor cells that proliferate and differentiate into retinal *** signaling,which is a fundamental mechanism known to drive cell-cell communication,is required to maintain Müller glia in a quiescent state in the undamaged retina,and repression of Notch signaling is necessary for Müller glia to reenter the cell *** dynamic regulation of Notch signaling following retinal damage also directs proliferation and neurogenesis of the Müller glia-derived progenitor cells in a robust regeneration *** contrast,mammalian Müller glia respond to retinal damage by entering a prolonged gliotic state that leads to additional neuronal death and permanent vision *** the dynamic regulation of Notch signaling in the zebrafish retina may aid efforts to stimulate Müller glia reprogramming for regeneration of the diseased human *** findings identified DeltaB and Notch3 as the ligand-receptor pair that serves as the principal regulators of zebrafish Müller glia *** addition,multi-omics datasets and functional studies indicate that additional Notch receptors,ligands,and target genes regulate cell proliferation and neurogenesis during the regeneration time ***,our understanding of Notch signaling during retinal regeneration is *** fully appreciate the complex regulation of Notch signaling that is required for successful retinal regeneration,investigation of additional aspects of the pathway,such as post-translational modification of the receptors,ligand endocytosis,and interactions with other fundamental pathways is *** we review various modes of Notch signaling regulation in the context of the vertebrate retina to put recent research in perspective and to identify open areas of inquiry.

关键词： differentiation gliosis Müller glia neuronal progenitor cell Notch signaling proliferation quiescence retinal development retinal regeneration zebrafish

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Live-cell imaging:new avenues to investigate retinal regeneration

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Neural regeneration Research 2017年第8期12卷 1210-1219页

作者： Manuela Lahne David R.Hyde Department of Biological Sciences University of Notre Dame Notre Dame IN USA

Sensing and responding to our environment requires functional neurons that act in concert. Neuronal cell loss resulting from degenerative diseases cannot be replaced in humans, causing a functional impairment to integrate and/or respond to sensory cues. In contrast, zebrafish（Danio rerio） possess an endogenous capacity to regenerate lost neurons. Here, we will focus on the processes that lead to neuronal regeneration in the zebrafish retina. Dying retinal neurons release a damage signal, tumor necrosis factor α, which induces the resident radial glia, the Müller glia, to reprogram and re-enter the cell cycle. The Müller glia divide asymmetrically to produce a Müller glia that exits the cell cycle and a neuronal progenitor cell. The arising neuronal progenitor cells undergo several rounds of cell divisions before they migrate to the site of damage to differentiate into the neuronal cell types that were lost. Molecular and immunohistochemical studies have predominantly provided insight into the mechanisms that regulate retinal regeneration. However, many processes during retinal regeneration are dynamic and require live-cell imaging to fully discern the underlying mechanisms. Recently, a multiphoton imaging approach of adult zebrafish retinal cultures was developed. We will discuss the use of live-cell imaging, the currently available tools and those that need to be developed to advance our knowledge on major open questions in the field of retinal regeneration.

关键词： multiphoton microscopy live-cell imaging zebrafish interkinetic nuclear migration tissue culture retinal regeneration Miiller glia neuronal progenitor cell differentiation phagocytosis

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Induction of Differentiation of Mesenchymal Stem Cells into retinal Pigment Epithelial Cells for retinal regeneration by Using Ciliary Neurotrophic Factor in Diabetic Rats

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Current Medical Science 2021年第1期41卷 145-152页

作者： Qing HUANG Yi DING Ji-guo YU Jing LI Yi XIANG Na TAO Department of Ophthalmology The Central Hospital of WuhanTongji Medical CollegeHuazhong University of Science and TechnologyWuhan430014China

Diabetic retinopathy(DR)is a common cause of blindness all over the *** marrow mesenchymal stem cells(BMSCs)have been considered as a promising strategy for retinal regeneration in the treatment of ***,the poor viability and low levels of BMSCs engraftment limit the therapeutic potential of *** present study aimed to examine the direct induction of BMSCs differentiation into the cell types related to retinal regeneration by using soluble cytokine ciliary neurotrophic factor(CNTF).We observed remarkably increased expression of cellular retinaldehyde-binding protein(CRALBP)and retinoid isomerohydrolase(RPE65)in BMSCs treated with CNTF in vitro,indicating the directional differentiation of BMSCs into the retinal pigment epithelium(RPE)cells,which are crucial for retinal *** vivo,the diabetic rat model was established by use of streptozotocin(STZ),and animals treated with BMSCs+CNTF exhibited better viability and higher delivery efficiency of the transplanted cells than those treated with BMSCs injection *** to the in-vitro result,treatment with BMSCs and CNTF combined led to the differentiation of BMSCs into beneficial cells(RPE cells),and accelerated retinal healing characterized by the activation of rod photoreceptor cells and phagocytosis function of RPE *** conclusion,CNTF contributes to the differentiation of BMSCs into RPE cells,which may help overcome the current stem cell therapy limitations in the field of retinal regeneration.

关键词： mesenchymal stem cells differentiation ciliary neurotrophic factor retinal regeneration retinal pigment epithelium(RPE)

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Absence of ephrin-A2/A3 increases retinal regenerative potential for Müller cells in Rhodopsin knockout mice

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Neural regeneration Research 2021年第7期16卷 1317-1322页

作者： Rui-Lin Zhu Yuan Fang Hong-Hua Yu Dong FChen Liu Yang Kin-Sang Cho Department of Ophthalmology Peking University First HospitalBeijingChina Schepens Eye Research Institute Massachusetts Eye and EarHarvard Medical SchoolBostonMAUSA Department of Ophthalmology and Visual Science Eye&ENT HospitalShanghai Medical CollegeFudan UniversityShanghaiChina Guangdong Eye Institute Department of OphthalmologyGuangdong Provincial People’s HospitalGuangdong Academy of Medical SciencesGuangzhouGuangdong ProvinceChina

Müller cells(MC) are considered dormant retinal progenitor cells in *** studies demonstrated ephrin-As act as negative regulators of neural progenitor cells in the retina and *** remains unclear whether the lack of ephrin-A2/A3 is sufficient to promote the neurogenic potential of *** we investigated whether the MC is the primary retinal cell type expressing ephrin-A2/A3 and their role on the neurogenic potential of Müller *** this study, we showed that ephrin-A2/A3 and their receptor EphA4 were expressed in retina and especially enriched in *** level of ephrin As/EphA4 expression increased as the retina matured that is correlated with the reduced proliferative and progenitor cell potential of ***, we investigated the proliferation in primary MC cultures isolated from wild-type and A2~(–/–) A3~(–/–) mice by 5-ethynyl-2′-deoxyuridine(EdU) *** detected a significant increase of EdU~+ cells in MC derived from A2~(–/–) A3~(–/–) ***, we investigated the role of ephrin-A2/A3 in mice undergoing photoreceptor degeneration such as Rhodopsin knockout(Rho~(–/–)) *** further evaluate the role of ephrin-A2/A3 in MC proliferation in vivo, EdU was injected intraperitoneally to adult wild-type, A2~(–/–) A3~(–/–), Rho~(–/–) and Rho~(–/–) A2~(–/–) A3~(–/–) mice and the numbers of EdU~+ cells distributed among different layers of the *** As/EphA4 expression was upregulated in the retina of Rho~(–/–) mice compared to the wild-type *** addition, cultured MC derived from ephrin-A2~(–/–) A3~(–/–) mice also expressed higher levels of progenitor cell markers and exhibited higher proliferation potential than those from wild-type ***, we detected a significant increase of EdU~+ cells in the retinas of adult ephrin-A2~(–/–) A3~(–/–) mice mainly in the inner nuclear layer;and these EdU~+ cells were co-localized with MC marker, cellular retinaldehyde-binding protein, suggesting some proliferating cells are from

关键词： endogenous stem cell EphA4 ephrin-A2 ephrin-A3 ephrins Müller cell photoreceptor cell regeneration retinal degeneration retinal regeneration retinal stem cell

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The role of Toll-like receptors in retinal ischemic diseases

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International Journal of Ophthalmology(English edition) 2016年第9期9卷 1343-1351页

作者： Wen-Qin Xu Yu-Sheng Wang Department of Ophthalmology Eye Institute of Chinese PLAXijing HospitalFourth Military Medical University

Toll-like receptors（TLRs） are commonly referred to a series of evolutionary conserved receptors which recognize and respond to various microbes and endogenous *** evidence has demonstrated that the expression of TLRs in the retina is regulated during retinal ischemic diseases,including ischemia-reperfusion injury,glaucoma,diabetic retinopathy（DR） and retinopathy of prematurity（ROP）.TLRs can be expressed in multiple cells in the retina,such as glial cells,retinal pigment epithelium（RPE）,as well as photoreceptor cells and endothelium *** of TLRs in retina could initiate a complex signal transduction cascade,induce the production of inflammatory cytokines and regulate the level of costimulatory molecules,which play prominent roles in the pathogenesis of retinal ischemic *** this review,we summarized current studies about the relationship between TLRs and ischemic retinopathy.A greater understanding of the effect of TLRs on ischemic injuries may contribute to the development of specific TLR targeted therapeutic strategies in these conditions.

关键词： Toll-like receptors retinopathy retinal ischemic diseases retinal regeneration

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Correlation between photoreceptor injury-regeneration and behavior in a zebrafish model

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Neural regeneration Research 2017年第5期12卷 795-803页

作者： Ya-jie Wang Shi-jiao Cai Jian-lin Cui Yang Chen Xin Tang Yu-hao Li Key Laboratory of Tumor Microenviroment and Neurovascular Regulation Nankai University School of Medicine Cataract Center Tianjin Eye Hospital

Direct exposure to intensive visible light can lead to solar retinopathy, including macular injury. The signs and symptoms include central scotoma, metamorphopsia, and decreased vision. However, there have been few studies examining retinal injury due to intensive light stimulation at the cellular level. Neural network arrangements and gene expression patterns in zebrafish photoreceptors are similar to those observed in humans, and photoreceptor injury in zebrafish can induce stem cell-based cellular regeneration. Therefore, the zebrafish retina is considered a useful model for studying photoreceptor injury in humans. In the current study, the central retinal photoreceptors of zebrafish were selectively ablated by stimulation with high-intensity light. retinal injury, cell proliferation and regeneration of cones and rods were assessed at 1, 3 and 7 days post lesion with immunohistochemistry and in situ hybridization. Additionally, a light/dark box test was used to assess zebrafish behavior. The results revealed that photoreceptors were regenerated by 7 days after the light-induced injury. However, the regenerated cells showed a disrupted arrangement at the lesion site. During the injury-regeneration process, the zebrafish exhibited reduced locomotor capacity, weakened phototaxis and increased movement angular velocity. These behaviors matched the morphological changes of retinal injury and regeneration in a number of ways. This study demonstrates that the zebrafish retina has a robust capacity for regeneration. Visual impairment and stress responses following high-intensity light stimulation appear to contribute to the alteration of behaviors.

关键词： nerve regeneration optic nerve injury light injury photoreceptor cell cell proliferation retinal regeneration light/dark test behavior zebrafish phototaxis immunohistochemistry in situ hybridization neural regeneration

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Ephrin-A2 and-A3 are negative regulators of the regenerative potential of Müller cells

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Chinese Medical Journal 2014年第19期127卷 3438-3442页

作者： Zhu Ruilin Cho Kin-Sang Chen Dong Feng Yang Liu Department of Ophthalmology Peking University First Hospital KeyLaboratory of Vision Loss and Restoration Ministry of EducationBeijing China Schepens Eye Research Institute Massachusetts Eye and EarHarvard Medical School Boston MA USA

Background In a previous study, we demonstrated that ephrin-A2 and -A3 negatively regulate the growth of neural progenitor cells in the central nervous system. Adult mice deficient in ephrin-A2 and -A3 （A2＋A3＋） displayed active ongoing neurogenesis throughout the brain, and mice deficient in ephrin-A3 alone showed increased proliferation of ciliary epithelium derived retinal stem cells. This study aimed to detect that the increase in proliferation and neurogenic potential of MOiler cells is influenced by the absence of ephrin-A2 and -A3. Methods We assessed the retinal and MOiler cell expression of ephrin-As and their receptor and neural progenitor cell markers by immunostaining and real-time PCR. We cultured purified primary MOiler cells derived from wild-type and A2＋A3＋ mice in a defined culture medium that enables trans-differentiation of Mu11er cells into retinal neurons. To evaluate proliferating MOiler cells in vivo, we injected 5＇-ethylnyl-2-deoxiuridine （EdU） intraperitoneally to adult mice. Results Expression of ephrin-A2/A3 and their receptor EphA4 were detected in the retinas of adult mice, with EphA4 expression particularly enriched in MOiler cells. MOiler cells of A2＋A3＋ mice exhibited significantly elevated expression of retinal progenitor cell markers, Pax6 and Chx10, when compared with those from wild-type mice. Moreover, a higher percentage of Mu11er cells of A2＋A3＋ mice trans-differentiated and became recoverin＋ and β-Ⅲ-tublin＋ in the culture than those from wild type mice. Strikingly, an increased number of EdU＋ retinal cells was detected in the retinas of adult A2＋A3＋ mice as compared with wild-type mice. Conclusions Ephrin-A2 and -A3 are negative regulators of the proliferative and neurogenic potentials of Mu11er cells. Manipulating ephrin-A signaling may thus represent a novel strategy for stimulating neuroregeneration from endogenous progenitors to participate in retinal repair in case of disease or damage.

关键词： Muller cell stem cell retinal regeneration

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