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Selective deletion of zinc transporter 3 in amacrine cells promotes retinal ganglion cell survival and optic nerve regeneration after injury

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Neural Regeneration Research 2023年第12期18卷 2773-2780页

作者： Zhe Liu Jingfei Xue Canying Liu Jiahui Tang Siting Wu Jicheng Lin Jiaxu Han Qi Zhang Caiqing Wu Haishun Huang Ling Zhao Yehong Zhuo Yiqing Li State Key Laboratory of Ophthalmology Zhongshan Ophthalmic CenterSun Yat-sen UniversityGuangdong Provincial Key Laboratory of Ophthalmology and Visual ScienceGuangzhouGuangdong ProvinceChina

Vision depends on accurate signal conduction from the retina to the brain through the optic nerve,an important part of the central nervous system that consists of bundles of axons originating from retinal ganglion cells.The mammalian optic nerve,an important part of the central nervous system,cannot regenerate once it is injured,leading to permanent vision loss.To date,there is no clinical treatment that can regenerate the optic nerve and restore vision.Our previous study found that the mobile zinc(Zn^(2+))level increased rapidly after optic nerve injury in the retina,specifically in the vesicles of the inner plexiform layer.Furthermore,chelating Zn^(2+)significantly promoted axonal regeneration with a long-term effect.In this study,we conditionally knocked out zinc transporter 3(ZnT3)in amacrine cells or retinal ganglion cells to construct two transgenic mouse lines(VGAT^(Cre)ZnT3^(fl/fl)and VGLUT2^(Cre)ZnT3^(fl/fl),respectively).We obtained direct evidence that the rapidly increased mobile Zn^(2+)in response to injury was from amacrine cells.We also found that selective deletion of ZnT3 in amacrine cells promoted retinal ganglion cell survival and axonal regeneration after optic nerve crush injury,improved retinal ganglion cell function,and promoted vision recovery.Sequencing analysis of reginal ganglion cells revealed that inhibiting the release of presynaptic Zn^(2+)affected the transcription of key genes related to the survival of retinal ganglion cells in postsynaptic neurons,regulated the synaptic connection between amacrine cells and retinal ganglion cells,and affected the fate of retinal ganglion cells.These results suggest that amacrine cells release Zn^(2+)to trigger transcriptomic changes related to neuronal growth and survival in reginal ganglion cells,thereby influencing the synaptic plasticity of retinal networks.These results make the theory of zinc-dependent retinal ganglion cell death more accurate and complete and provide new insights into the complex interactions between retinal cell networks.

关键词： axonal regeneration conditional knockout neurotransmitter optic nerve injury presynaptic neuron retinal network synaptic connection synaptic vesicles visual acuity zinc transporter 3

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Adult neurogenesis in the primate hippocampus

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Zoological Research 2023年第2期44卷 315-322页

作者： Ye Li Na-Na Xu Zhao-Zhe Hao Sheng Liu State Key Laboratory of Ophthalmology Zhongshan Ophthalmic CenterSun Yat-sen UniversityGuangdong Provincial Key Laboratory of Ophthalmology and Visual ScienceGuangzhouGuangdong 510060China Guangdong Province key laboratory of Brain Function and Disease GuangzhouGuangdong 510080China

Adult hippocampal neurogenesis(AHN) is crucial for learning,memory,and emotion.Deficits of AHN may lead to reduced cognitive abilities and neurodegenerative disorders,such as Alzheimer’s disease.Extensive studies on rodent AHN have clarified the developmental and maturation processes of adult neural stem/progenitor cells.However,to what extent these findings apply to primates remains controversial.Recent advances in next-generation sequencing technologies have enabled in-depth investigation of the transcriptome of AHN-related populations at single-cell resolution.Here,we summarize studies of AHN in primates.Results suggest that neurogenesis is largely shared across species,but substantial differences also exist.Marker gene expression patterns in primates differ from those of rodents.Compared with rodents,the primate hippocampus has a higher proportion of immature dentate granule cells and a longer maturation period of newly generated granule cells.Future research on species divergence may deepen our understanding of the mechanisms underlying adult neurogenesis in primates.

关键词： Neural precursor cells Immature granule cells Sc-RNA-seq

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Protective effect of ginsenoside Rg1 on 661W cells exposed to oxygen-glucose deprivation/reperfusion via keap1/nrf2 pathway

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International Journal of ophthalmology(English edition) 2023年第7期16卷 1026-1033页

作者： Ming Zhou Xin-Qi Ma Yi-Yu Xie Jia-Bei Zhou Xie-Lan Kuang Huang-Xuan Shen Chong-De Long State Key Laboratory of Ophthalmology Zhongshan Ophthalmic CenterSun Yat-sen UniversityGuangdong Provincial Key Laboratory of Ophthalmology and Visual ScienceGuangzhou 510060China

AIM:To construct an in vitro model of oxygen-glucose deprivation/reperfusion(OGD/R)induced injury to the optic nerve and to study the oxidative damage mechanism of ischemia-reperfusion(I/R)injury in 661W cells and the protective effect of ginsenoside Rg1.METHODS:The 661W cells were treated with different concentrations of Na2S2O4 to establish OGD/R model in vitro.Apoptosis,intracellular reactive oxygen species(ROS)levels and superoxide dismutase(SOD)levels were measured at different time points during the reperfusion injury process.The injury model was pretreated with graded concentrations of ginsenoside Rg1.Real-time polymerase chain reaction(PCR)was used to measure the expression levels of cytochrome C(cyt C)/B-cell lymphoma-2(Bcl2)/Bcl2 associated protein X(Bax),heme oxygenase-1(HO-1),caspase9,nuclear factor erythroid 2-related factor 2(nrf2),kelch-like ECH-associated protein 1(keap1)and other genes.Western blot was used to detect the expression of nrf2,phosphorylated nrf2(pnrf2)and keap1 protein levels.RESULTS:Compared to the untreated group,the cell activity of 661W cells treated with Na2S2O4 for 6 and 8h decreased(P<0.01).Additionally,the ROS content increased and SOD levels decreased significantly(P<0.01).In contrast,treatment with ginsenoside Rg1 reversed the cell viability and SOD levels in comparison to the Na_(2)S_(2)O_(4)treated group(P<0.01).Moreover,Rg1 reduced the levels of caspase3,caspase9,and cyt C,while increasing the Bcl2/Bax level.These differences were all statistically significant(P<0.05).Western blot analysis showed no significant difference in the protein expression levels of keap1 and nrf2 with Rg1 treatment,however,Rg1 significantly increased the ratio of pnrf2/nrf2 protein expression compared to the Na_(2)S_(2)O_(4)treated group(P<0.001).CONCLUSION:The OGD/R process is induced in 661W cells using Na_(2)S_(2)O_(4).Rg1 inhibits OGD/R-induced oxidative damage and alleviates the extent of apoptosis in 661W cells through the keap1/nrf2 pathway.These results suggest a potential protective ef

关键词： oxygen-glucose deprivation/reoxygenation ginsenoside Rg1 oxidative stress phosphorylated nrf2

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多孔复合水凝胶负载角膜缘干细胞重建眼表（英文）

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Science China Materials 2024年

作者：王付燕许悦荷石德鹏程钧赵龙祁霞周庆军谢立信 Shandong Provincial key laboratory of ophthalmology Eye Institute of Shandong First Medical University state key laboratory Cultivation Base School of ophthalmology

干细胞疗法是一种用于重建角膜缘干细胞缺乏症(LSCD)患者眼表的新技术.然而,目前报道的干细胞疗法治疗LSCD仅局限于二维(2D)培养体系,不能模拟干细胞在体内生存的原生微环境.本研究基于多孔明胶甲基丙烯酰(GelMA)/丝素蛋白甲基丙烯酸缩...

干细胞疗法是一种用于重建角膜缘干细胞缺乏症(LSCD)患者眼表的新技术.然而,目前报道的干细胞疗法治疗LSCD仅局限于二维(2D)培养体系,不能模拟干细胞在体内生存的原生微环境.本研究基于多孔明胶甲基丙烯酰(GelMA)/丝素蛋白甲基丙烯酸缩水甘油酯(SilM A),制备了一种可注射的光交联水凝胶用于负载角膜缘干细胞(LSCs)以重建眼表.多孔GelMA/SilMA水凝胶具有优异的透明度和良好的粘附性.更重要的是,该水凝胶保持了兔LSCs的活力,支持了兔LSCs的粘附,并促进了它们在体外3D培养环境中的迁移.蛋白质组学分析证实,多孔GelMA/SilMA水凝胶3D细胞培养体系能够保持LSCs的自我更新和分化能力.此外,组织学分析表明,负载LSCs的多孔GelMA/SilMA水凝胶能够有效的促进LSCD再上皮化.多孔水凝胶能够简化连续细胞培养的相关操作,以构建具有细胞均匀分布的眼表.这些发现为干细胞疗法和再生医学用于眼表重建提供了有价值的见解.

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Structural insights into the CRISPR-Cas-associated ribonuclease activity of Staphylococcus epidermidis Csm3 and Csm6: a step for the development of novel genome editing tool

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Science Bulletin 2018年第11期63卷 672-674页

作者： Feng Gu School of ophthalmology and Optometry Eye Hospital Wenzhou Medical University State Key Laboratory Cultivation Base and Key Laboratory of Vision Science Ministry of Health and Zhejiang Provincial Key Laboratory of Ophthalmology and Optometry

CRISPR-Cas systems has been developed as genome editing tools,which are originally discovered as sequence-specific defense system utilized by bacteria and archaea to fight with the invasion by foreign nucleic acids such as phages or plasmids.According to the current classification groups of CRISPR-Cas system,Type I,Type III and Type IV comprise the Class 1 systems for their function through multisubunit Cas protein complex to interference the targeted nucleic acids.In contrast,Class 2 systems,consisting of

关键词：编辑工具染色体葡萄球菌结构防卫系统噬菌体

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Entrainment of Astrocytic and Neuronal Ca^(2+) Population Dynamics During Information Processing of Working Memory in Mice

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Neuroscience Bulletin 2022年第5期38卷 474-488页

作者： Zhu Lin Feng You Ting Li Yijia Feng Xinyue Zhao Jingjing Yang Zhimo Yao Ying Gao Jiang-Fan Chen The Molecular Neuropharmacology laboratory and the EyeBrain Research Center State Key Laboratory of OphthalmologyOptometry and Visual ScienceSchool of Optometry and Ophthalmology and Eye HospitalWenzhou Medical UniversityWenzhou 325035China

Astrocytes are increasingly recognized to play an active role in learning and memory,but whether neural inputs can trigger event-specific astrocytic Ca^(2+)dynamics in real time to participate in working memory remains unclear due to the difficulties in directly monitoring astrocytic Ca^(2+)dynamics in animals performing tasks.Here,using fiber photometry,we showed that population astrocytic Ca^(2+)dynamics in the hippocampus were gated by sensory inputs(centered at the turning point of the T-maze)and modified by the reward delivery during the encoding and retrieval phases.Notably,there was a strong inter-locked and antagonistic relationship between the astrocytic and neuronal Ca^(2+)dynamics with a 3-s phase difference.Furthermore,there was a robust synchronization of astrocytic Ca^(2+)at the population level among the hippocampus,medial prefrontal cortex,and striatum.The inter-locked,bidirectional communication between astrocytes and neurons at the population level may contribute to the modulation of information processing in working memory.

关键词： Working memory Ca^(2+)dynamics Astrocyte Neuron Fiber photometry Hippocampus

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Natural polymer-derived photocurable bioadhesive hydrogels for sutureless keratoplasty

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Bioactive Materials 2022年第2期7卷 196-209页

作者： Xuan Zhao Saiqun Li Xinyue Du Weihua Li Qian Wang Dalian He Jin Yuan State Key Laboratory of Ophthalmology Zhongshan Ophthalmic CenterSun Yat-sen UniversityGuangzhou510623China

Keratoplasty is the gold standard treatment for visual impairment caused by corneal damage.The use of suturing as the bonding method is the source of many complications following keratoplasty.Currently available corneal adhesives do not have both adequate adhesive strength and acceptable biocompatibility.Herein,we developed a photocurable bioadhesive hydrogel which was composed of gelatin methacryloyl and oxidized dextran for sutureless keratoplasty.The bioadhesive hydrogel exhibited high light transmittance,resistance to enzymatic degradation and excellent biocompatibility.It also had higher adhesive strength than commercial adhesives(fibrin glue).In a rabbit model of lamellar keratoplasty,donor corneal grafts could be closely bonded to the recipient corneal bed and remained attached for 56 days by using of this in situ photopolymerized bioadhesive hydrogel.The operated cornea maintained transparent and noninflamed.Sutureless keratoplasty using bioadhesive hydrogel allowed rapid graft re-epithelialization,typically within 7 days.In vivo confocal microscopic and histological evaluation of the operated cornea did not show any apparent abnormalities in terms of corneal cells and ultrastructure.Thus,this bioadhesive hydrogel is exhibited to be an appealing alternative to sutures for keratoplasty and other corneal surgeries.

关键词： Bioadhesive hydrogels Photocurable Cornea repair Sutureless keratoplasty

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Inhibition of Foxp4 Disrupts Cadherin-based Adhesion of Radial Glial Cells,Leading to Abnormal Differentiation and Migration of Cortical Neurons in Mice

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Neuroscience Bulletin 2023年第7期39卷 1131-1145页

作者： Xue Li Shimin Zou Xiaomeng Tu Shishuai Hao Tian Jiang Jie-Guang Chen School of ophthalmology and Optometry and Eye Hospital Wenzhou Medical UniversityWenzhou 325027China state key laboratory of Optometry Ophthalmology and Vision Scienceand Zhejiang Provincial Key Laboratory of Optometry and OphthalmologyWenzhou 325027China Research Center for Translational Medicine the Afliated Wenling Hospital of Wenzhou Medical UniversityWenling 317500China

Heterozygous loss-of-function variants of FOXP4 are associated with neurodevelopmental disorders(NDDs)that exhibit delayed speech development,intellectual disability,and congenital abnormalities.The etiology of NDDs is unclear.Here we found that FOXP4 and N-cadherin are expressed in the nuclei and apical end-feet of radial glial cells(RGCs),respectively,in the mouse neocortex during early gestation.Knockdown or dominant-negative inhibition of Foxp4 abolishes the apical condensation of N-cadherin in RGCs and the integrity of neuroepithelium in the ventricular zone(VZ).Inhibition of Foxp4 leads to impeded radial migration of cortical neurons and ectopic neurogenesis from the proliferating VZ.The ectopic differentiation and deficient migration disappear when N-cadherin is over-expressed in RGCs.The data indicate that Foxp4 is essential for N-cadherin-based adherens junctions,the loss of which leads to periventricular heterotopias.We hypothesize that FOXP4 variant-associated NDDs may be caused by disruption of the adherens junctions and malformation of the cerebral cortex.

关键词： Neurogenesis Radial migration Radial glial cell Ventricular zone N-cadherin

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Comparisons of ganglion cell-inner plexiform layer loss patterns and its diagnostic performance between normal tension glaucoma and primary open angle glaucoma: a detailed, severity-based study

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International Journal of ophthalmology(English edition) 2020年第1期13卷 71-78页

作者： Xiao-Yu Xu Kun-Bei Lai Hui Xiao Yi-Quan Lin Xin-Xing Guo Xing Liu State Key Laboratory of Ophthalmology Zhongshan Ophthalmic Center Sun Yat-sen University

AIM: To evaluate the patterns of macular ganglion cell-inner plexiform layer(GCIPL) loss in normal tension glaucoma(NTG) and primary open angle glaucoma(POAG) in a detailed, disease severity-matched way;and to assess the diagnostic capabilities of GCIPL thickness parameters in discriminating NTG or POAG from normal subjects.METHODS: A total of 157 eyes of 157 subjects, including 57 normal eyes, 51 eyes with POAG and 49 eyes with NTG were enrolled and strictly matched in age, refraction, and disease severity between POAG and NTG groups. The average, minimum, superotemporal, superior, superonasal, inferonasal, inferior, and inferotemporal GCIPL thickness, and the average, superior, temporal, inferior, and nasal retinal nerve fiber layer(RNFL) thickness were obtained by Cirrus optical coherence tomography(OCT). The diagnostic capabilities of OCT parameters were assessed by area under receiver operating characteristic(AUROC) curves. RESULTS: Among all the OCT thickness parameters, no statistical significant difference between NTG group and POAG group was found(all P>0.05). In discriminating NTG or POAG from normal subjects, the average and inferior RNFL thickness, and the minimum GCIPL thickness had better diagnostic capabilities. There was no significant difference in AUROC curve between the best GCIPL thickness parameter(minimum GCIPL) and the best RNFL thickness parameter in discriminating NTG(inferior RNFL;P=0.076) and indiscriminating POAG(average RNFL;P=0.913) from normal eyes.CONCLUSION: Localized GCIPL loss, especially in the inferior and inferotemporal sectors, is more common in NTG than in POAG. Among all the GCIPL thickness parameters, the minimum GCIPL thickness has the best diagnostic performance in differentiating NTG or POAG from normal subjects, which is comparable to that of the average and inferior RNFL thickness.

关键词： normal tension glaucoma primary open angle glaucoma spectral domain optical coherence tomography ganglion cell-inner plexiform layer thickness pattern

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Rare variants in novel and known genes associated with primary angle closure glaucoma based on whole exome sequencing of 549 probands

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Journal of Genetics and Genomics 2019年第7期46卷 353-357页

作者： Wenmin Sun Xueshan Xiao Shiqiang Li Jiamin Ouyang Xueqing Li Xiaoyun Jia Xing Liu Qingjiong Zhang State Key Laboratory of Ophthalmology Zhongshan OphthalmicCenter Sun Yat-sen University

Primary angle closure glaucoma(PACG)is one of the most common causes of irreversible blindness in Asia and about 80%of PACG are present in Asia(Song et al.,2017).Genetics plays an important role in the development of PACG(Gramer et al.,2014).Recently,eight common SNPs were found to be strongly associated with PACG(Vithana et al.,2012;Khor et al.,2016)and mutations in MYRF cause high hyperopia accompanied with PACG(Xiao et al.,2019).

关键词： irreversible blindness hyperopia

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