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Overexpressing NeuroD1 reprograms Müller cells into various types of retinal neurons

Neural Regeneration Research 2023年第5期18卷 1124-1131页

作者： Di Xu Li-Ting Zhong Hai-Yang Cheng Zeng-Qiang Wang Xiong-Min Chen Ai-Ying Feng Wei-Yi Chen Gong Chen Ying Xu Guangdong-Hongkong-Macao Institute of CNS Regeneration Key Laboratory of CNS Regeneration(Ministry of Education)Jinan UniversityGuangzhouGuangdong ProvinceChina Co-Innovation Center of Neuroregeneration Nantong UniversityNantongJiangsu ProvinceChina

The onset of retinal degenerative disease is often associated with neuronal loss. Therefore, how to regenerate new neurons to restore vision is an important issue. NeuroD1 is a neural transcription factor with the ability to reprogram brain astrocytes into neurons in vivo. Here, we demonstrate that in adult mice, NeuroD1 can reprogram Müller cells, the principal glial cell type in the retina, to become retinal neurons. Most strikingly, ectopic expression of NeuroD1 using two different viral vectors converted Müller cells into different cell types. Specifically, AAV7 m8 GFAP681::GFP-ND1 converted Müller cells into inner retinal neurons, including amacrine cells and ganglion cells. In contrast, AAV9 GFAP104::ND1-GFP converted Müller cells into outer retinal neurons such as photoreceptors and horizontal cells, with higher conversion efficiency. Furthermore, we demonstrate that Müller cell conversion induced by AAV9 GFAP104::ND1-GFP displayed clear dose-and time-dependence. These results indicate that Müller cells in adult mice are highly plastic and can be reprogrammed into various subtypes of retinal neurons.

关键词： amacrine cell ganglion cell horizontal cell in vivo reprogramming Müller cell NeuroD1 photoreceptor regeneration retina retinal degeneration

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Modification of retinal horizontal cell morphology during photoreceptor degeneration in the RCS rats

Modification of retinal horizontal cell morphology during ph...

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中国神经科学学会第四次会员代表大会暨第八届全国学术会议

作者： Qiang ZENG~1,Hai-Tao ZHANG~2,Zhi-Ping LIU~1,Yi WANG~2,Zheng-Qin YIN~(2*),Shi-Gang HE~(1,*) 1 State Key Laboratory of Brain and Cognitive Sciences,Institute of Biophysics,Chinese Academy of Sciences,Beijing 100101,China 2 Southwest Eye Hospital,Chongqing 400038,China

＜正＞Objective The Royal College of Surgeons（RCS） rats undergo a sustained degeneration of photoreceptors from the retinal pigment epithelial cell defect.As one kind of intemeurons,horizontal cells connect to cone and rod photoreceptors directly in the outer plexiform layer,which may be influenced by the loss of photoreceptors.In order to examine the remodeling of the surviving secondary neurons,here we investigated the lamination and morphology of horizontal cells （HCs） during photoreceptor degeneration in the RCS rats.Methods RCS rats were sampled at different degeneration stages and compared with age-matched control data.Both immunostaining of the cryosections and DiI labeling of the wholemount retinas were carried out to examine soma,dendritic and axonal morphology.Images were acquired by confocol microscopy and analyzed with ImageJ.Results The staining of photoreceptors shows that at P35,the beginning of the photoreceptor degeneration,the loss of photoreceptors is not significant and the morphology of the HCs is not changed. At P45 the loss of photoreceptors is obvious in the RCS rats.But the lamination and morphology of the HCs are not significantly changed.At 2 months,with 20%or less photoreceptors left,the area of dendrites and axon terminals are both reduced significantly and dendritic arbors become much simpler.But the lamination of the HCs is still normal at this stage. At 3 months,nearly all the photoreceptors are degenerated.The dendritic arbor area is reduced and some of the dendrites tangle with each other;the axon terminals show a very simple morphology with only some main branches.Some axon terminals show ectropic branches and the lamination is disrupted.Conclusion The morphology of HCs changes with the loss of the photoreceptors in the RCS rats.Both the dendrites and axon terminals become simpler and smaller,but the modification is slower than the loss of photoreceptors.The lamination of the HC neurites is normal in the outer plexiform layer even at 2 months;the extropic neurites into the outer nu

关键词： RCS rat horizontal cell DiI labeling

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Morphology change of secondary neuron under fast and slow photoreceptor degeneration

Morphology change of secondary neuron under fast and slow ph...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Qiang ZENG~1,Hai-Tao ZHANG~2,Yan YIN~1,Zhi-Ping LIU~1,Yi WANG~2,Zheng-Qin YIN~(2,*),Shi-Gang HE~(1,*) 1 State Key Laboratory of Brain and Cognitive Sciences,Institute of Biophysics,Chinese Academy of Sciences,15 Da- tun Road,Beijing 100101,China 2 Southwest Eye Hospital,Chongqing 400038,China

＜正＞Objective To investigate the influence of photoreceptor degeneration on the morphology of secondary neurons（bipolar and horizontal cells） in the retina of Royal College of Surgeons（RCS） rat and light damaged（LD） albino mouse.Methods RCS rats and LD mice retinas were sampled at different degeneration stages and compared with wild type（wt） retinas;Both immunohistochemistry and Dil labeling methods were used to examine lamination and morphology of the secondary neurons.Images were acquired by confocal microscopy and analyzed with ImageJ. Results Gradual loss of photoreceptors and ribbon synapses was detected from postnatal day 30（P30） to P90 in the RCS rat retina.Rod bipolar cells and horizontal cells kept the neurite lamination in outer plexiform layer（OPL） at P60 when 20%photoreceptors left;horizontal cells maintained their morphology at P45 when 50%photoreceptors left.LD induced fast loss of photoreceptors and ribbon synapses in albino mouse retina.Lamination and morphology disruption of the bipolar cells happened at 1 day after light damage（DALD）.Morphology change of horizontal cells appeared at 5 DALD.Conclusion Structure alternation of secondary neurons in RCS rat retina appears later than photoreceptor loss during the slow degeneration process.But in LD mouse retina the alternation appears early following fast photoreceptor loss,indicating the morphology change could happen fast and the residual photoreceptors maintain the structure of secondary neurons.

关键词： retina Royal College of Surgeons light damage horizontal cell bipolar cell

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Modulation of inward-rectifying K+ current by activation of NMDA receptors on carp retinal horizontal cells

Modulation of inward-rectifying K+ current by activation of ...

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中国神经科学学会第四次会员代表大会暨第八届全国学术会议

作者： Yan SUN,Xiao-Dong JIANG,Lei XIAO,Hai-Qing GONG,Pei-Ji LIANG~* School of Life Sciences and Biotechnology,Shanghai Jiao Tong University,Shanghai 200240,China

Objective Recently,NMDA receptors are reported to be expressed in carp retinal horizontal cells,but the function of these NMDA receptors has yet to be further determined.Activation of NMDA receptors results in modulation of many voltage-gated ion channels,therefore,we wish to ask whether activation of these NMDA receptors could alter the inward-rectifying K current(Ikir),which is also identified in the retinal horizontal cells.Methods H1 horizontal cells were isolated from carp retina and currents were recorded by patch-clamp.All the drugs were briefly delivered by the superfusion system(DAD-12,ALA Scientific Instruments,USA).Tetrodotoxin(TTX,10μM) and nifedipine(100μM) were used to block voltage-gated Na channel and L-type Ca channel respectively in all the experiments.Results(1) IKir was measured with step commands to potentials more hyperpolarized than -80 mV on H1 horizontal cells and this inward current could be blocked by 1 mM Cs+.(2) Furthermore,expose to NMDA was associated with an observable reduction of IKir on H1 horizontal cells.Conclusion Activation of NMDA receptors inhibits the IKir on carp retinal horizontal cells.

关键词： inward-rectifying K+ current NMDA receptors horizontal cell patch-clamp

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