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Use of a tissue clearing technique combined with retrograde trans-synaptic viral tracing to evaluate changes in mouse retinorecipient brain regions following optic nerve crush

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Neural Regeneration Research 2023年第4期18卷 913-921页

作者： Zong-Yi Zhan Yi-Ru Huang Lu-Wei Zhao Ya-Dan Quan Zi-Jing Li Di-Fang Sun Ya-Li Wu Hao-Yuan Wu Zi-Tian Liu Kai-Li Wu Yu-Qing Lan Min-Bin Yu Department of Ophthalmology Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene RegulationSun Yat-sen Memorial HospitalSun Yat-sen UniversityGuangzhouGuangdong ProvinceChina State Key Laboratory of Ophthalmology Zhongshan Ophthalmic CenterSun Yat-sen UniversityGuangzhouGuangdong ProvinceChina Light Innovation Technology Ltd. Hong Kong Special Administrative RegionChina Department of Ophthalmology&Visual Science Shanghai Medical CollegeFudan UniversityShanghaiChina Department of Ophthalmology Eye&ENT HospitalFudan UniversityShanghaiChina Department of Ophthalmology The First Affiliated Hospital of USTCDivision of Life Sciences and MedicineUniversity of Science and Technology of ChinaHefeiAnhui ProvinceChina

Successful establishment of reconnection between retinal ganglion cells and retinorecipient regions in the brain is critical to optic nerve ***,morphological assessments of retinorecipient regions are limited by the opacity of brain *** this study,we used an innovative tissue cleaning technique combined with retrograde trans-synaptic viral tracing to observe changes in retinorecipient regions connected to retinal ganglion cells in mice after optic nerve ***,we performed light-sheet imaging of whole brain tissue after a clearing *** found that pseudorabies virus 724(PRV724)mostly infected retinal ganglion cells,and that we could use it to retrogradely trace the retinorecipient regions in whole tissue-cleared ***,PRV724-traced neurons were more widely distributed compared with data from previous *** found that optic nerve injury could selectively modify projections from retinal ganglion cells in the hypothalamic paraventricular nucleus,intergeniculate leaflet,ventral lateral geniculate nucleus,central amygdala,basolateral amygdala,Edinger-Westphal nucleus,and oculomotor nucleus,but not the superior vestibular nucleus,red nucleus,locus coeruleus,gigantocellular reticular nucleus,or facial nerve *** findings demonstrate that the tissue clearing technique,combined with retrograde trans-synaptic viral tracing,can be used to objectively and comprehensively evaluate changes in mouse retinorecipient regions that receive projections from retinal ganglion cells after optic nerve ***,our approach may be useful for future estimations of optic nerve injury and regeneration.

关键词： histology image analysis light-sheet imaging optic nerve crush pseudorabies virus retinal ganglion cells three-dimensional imaging tissue clearing viral tracing whole brain study

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Altered Energy Metabolism During Early optic nerve crush Injury:Implications of Warburg-Like Aerobic Glycolysis in Facilitating Retinal Ganglion Cell Survival

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Neuroscience Bulletin 2020年第7期36卷 761-777页

作者： Jingyi Zhu Ping Li Yuan-Guo Zhou Jian Ye Department of Ophthalmology Army Medical Center of the People's Liberation Army(PLA)Army Medical UniversityChongqing 400042China Molecular Biology Center State Key Laboratory of TraumaBumand Combined InjuryResearch Institute of SurgeryArmy Medical Center of the PLAArmy Medical UniversityChongqing 400042China

Neurons,especially axons,are metabolically demanding and energetically vulnerable during ***,the exact energy budget alterations that occur early after axon injury and the effects of these changes on neuronal survival remain *** a classic mouse model of optic nerve-crush injury,we found that traumatized optic nerves and retinas harbor the potential to mobilize two primary energetic machineries,glycolysis and oxidative phosphorylation,to satisfy the robustly increased adenosine triphosphate(ATP) *** exploration of metabolic activation showed that mitochondrial oxidative phosphorylation was amplified over other pathways,which may lead to decreased retinal ganglion cell(RGC) survival despite its supplement to ATP *** set enrichment analysis of a microarray(GSE32309) identified significant activation of oxidative phosphorylation in injured retinas from wild-type mice compared to those from mice with deletion of phosphatase and tensin homolog(PTEN),while PTEN-/-mice had more robust RGC ***,we speculated that the oxidation-favoring metabolic pattern after optic nervecrush injury could be adverse for RGC *** redirecting metabolic flux toward glycolysis(magnifying the Warburg effect) using the drug meclizine,we successfully increased RGC ***,we provide novel insights into a potential bioenergetics-based strategy for neuroprotection.

关键词： optic nerve crush ATP Glycolysis Oxidative phosphorylation RGC survival

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Valproate reduces retinal ganglion cell apoptosis in rats after optic nerve crush

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Neural Regeneration Research 2023年第7期18卷 1607-1612页

作者： Feng Pan Dan Hu Li-Juan Sun Qian Bai Yu-Sheng Wang Xu Hou Eye Institute of Chinese PLA and Department of Ophthalmology Xijing HospitalFourth Military Medical UniversityXi’anShaanxi ProvinceChina

The retinal ganglion cells of the optic nerve have a limited capacity for self-repair after *** is a histone deacetylase inhibitor and multitarget drug,which has been demonstrated to protect retinal *** this study,we established rat models of optic nerve-crush injury and injected valproate into the vitreous cavity immediately after *** evaluated changes in the ultrastructure morphology of the endoplasmic reticulum of retinal ganglion cells over time via transmission electron *** and western blot assay revealed that valproate upregulated the expression of the endoplasmic reticulum stress marker glucose-regulated protein 78 and downregulated the expression of transcription factor C/EBP homologous protein,phosphorylated eukaryotic translation initiation factor 2α,and caspase-12 in the endoplasmic reticulum of retinal ganglion *** findings suggest that valproate reduces apoptosis of retinal ganglion cells in the rat after optic nerve-crush injury by attenuating phosphorylated eukaryotic translation initiation factor 2α-C/EBP homologous protein signaling and caspase-12 activation during endoplasmic reticulum *** findings represent a newly discovered mechanism that regulates how valproate protects neurons.

关键词： apoptosis C/EBP homologous protein caspase-12 endoplasmic reticulum glucose-regulated protein 78 optic nerve crush phosphorylated eukaryotic translation initiation factor 2α retinal ganglion cells unfolded protein response valproate

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Protective effects of Achyranthes bidentata polypeptides on retinal ganglion cells post-optic nerve crush in rats

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Neural Regeneration Research 2011年第15期6卷 1164-1168页

作者： Nan Hu Qi Zhao Fangling Zhang Junfang Zhang Xiaosong Gu Medical School of Soochow University Suzhou 215123 Jiangsu Province China Department of Ophthalmology Affiliated Hospital of Nantong University Nantong 226001 Jiangsu Province China Jiangsu Key Laboratory of Neuroregeneration Nantong University Nantong 226001 Jiangsu Province China

Achyranthes bidentata polypeptides（ABPP） have been reported to inhibit apoptosis of retinal ganglion cells（RGCs）.The present study investigated the protective effects of ABPP on RGCs in a rat model of optic nerve *** prolonged injury time,RGC densities were gradually ***（5 μg） significantly increased RGC densities and upregulated growth associated protein 43 expression in rats with optic nerve *** demonstrate that ABPP can protect RGCs and promote axonal growth after optic nerve crush.

关键词： Achyranthes bidentata polypeptides optic nerve crush retinal ganglion cells growth associated protein 43 neural regeneration

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Erythropoietin upregulates growth associated protein-43 expression and promotes retinal ganglion cell axonal regeneration in vivo after optic nerve crush

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Neural Regeneration Research 2012年第4期7卷 295-301页

作者： Haibo Tan Xin Kang Yisheng Zhong Xi Shen Yu Cheng Qin Jiao Lianfu Deng Department of Ophthalmology Ruijin Hospital Shanghai Jiao Tong University School of Medicine Shanghai 200025 China Department of Clinical Pharmacology Changhai Hospital Affiliated to Second Mifitary Medical University Shanghai 200433 China Shanghai Institute of Traumatology and Orthopedics Shanghai 200025 China

In this study, we established a rat model of optic nerve crush to explore the effects of erythropoietin on retinal ganglion cell axonal regeneration. At 15 days after injury in erythropoietin treated rats, retinal ganglion cell densities in regions corresponding to the 1/6, 3/6 and 5/6 ratios of the retinal radius were significantly increased. In addition, the number of growth associated protein-43 positive axons was significantly increased at different distances （50, 250 and 500 pm） from the crush site after erythropoietin treatment. Erythropoietin significantly increased growth associated protein-43 protein levels in the retina after crush injury, as determined by westem blot and immunofluorescence analysis. These results demonstrate that erythropoietin protects injured retinal ganglion cells and promotes axonal regeneration.

关键词： erythropoietin retinal ganglion cells axonal regeneration optic nerve crush neural regeneration

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Protective Effect of Paeoniflorin against optic nerve crush

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2007年第6期27卷 650-652页

作者：李敬熊新春刘云海 Department of Pharmacy Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Department of Ophthalmology Union Hospital Tongji Medical College Huazhong University of Science and Technology

In order to evaluate the efficacy of traditional paeonia extract paeoniflorin against optic nerve crush, 16 Brown Norway rats were divided into two groups at random, with 8 rats in each group. In paeoniaflorin-treated group, 2 mg paeoniaflorin （total volum： 1 mL） was injected into rat＇s peritoneum one time a day for a period of 8 days. Rats in untreated group were given a single dose of vehicle. The optic nerve was crushed by a special forceps for 30 s in the left eye and a sham procedure was performed in the right eye on the 2nd day after the first injection. The retrograde fluorogold labeling of ganglion cells was conducted 5 days after optic nerve crush. The whole retina was flat-mounted thereafter. The ganglion cells that survived the crush were counted under fluorescent microscope by using an automatic counting software. As compared with the contralateral eye, the survival rate of ganglion cells in the left eye increased from 40.22% to 64.53% with a significant difference found between them （t=2.55, P=0.023）. The results suggested that the paeonia extract paeoniflorin possessed a protective effect against optic nerve crush.

关键词： paeoniflorin optic nerve crush retina ganglion cell retrograde labeling

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Deprivation of form vision induces myopia in mice with optic nerve crush

Deprivation of form vision induces myopia in mice with optic...

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中国神经科学学会第十届全国学术会议

作者： Xiao-Hua WU Yun-Yun LI Ping-Ping ZHANG Shi-Jun WENG Xiong-Li YANG Yong-Mei ZHONG Institute of Neurobiology Institutes of Brain Science and State Key Laboratory of Medical NeurobiologyFudan University

Objective Deprivation of form vision causes excessive axial eye growth and a myopic refractive error in juveniles of several ***,the brain may also be involved in the control of eye *** this work,we investigated whether the retina-brain link is necessary for form-deprivation myopia in mice with optic nerve crush（ONC）.Methods Eye refraction was measured by an automated eccentric infrared photorefractor and ocular components were measured by optical coherence tomography（OCT）.C57BL/6 mice（21-days-old） were monocularly deprived of form vision with white translucent occluders for 4 *** assess the role of brain in form-deprivation myopia,the right optic nerve was crushed in mice（18-days-old） 3 days before the right eyes were occluded for 4 weeks *** The occluded eyes with intact optic nerve showed a significant myopic shift（-5.55±1.83 D）,relative to the contralateral fellow eyes,after 4-week visual *** mice with ONC（but without occluders）,the injured eyes became either myopic or hyperopic,compared to the fellow *** the group with myopic-shift（-5.73±4.81D）,the injured eyes exhibited a significant decrease in both lens and retinal thickness（P ＜0.05）.In the group with hyperopic-shift（4.59±3.03 D）,the injured eyes showed a decrease in corneal and retinal dnckness（P ＜0.05）.In mice with ONC,the occluded eyes all became myopic,compared to the fellow eyes,with a myopic shift（-9.69±4.17 D） even larger than that（-5.55±1.83 D）observed in occluded eyes in mice with intact optic ***,these occluded eyes exhibited a significant increase in axial length and vitreous chamber depth（P ＜0.05）,and a decrease in retinal thickness（P ＜0.001）,lens thickness and corneal radius of curvature（P ＜0.05）.Conclusion Intact eye-brain link is indispensable for normal ocular growth and refractive *** centers are involved in the development of myopia due to form deprivation.

关键词： mouse form deprivation myopia optic nerve crush eye growt

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Quantitative investigation of optic nerve remyelination after crush in adult zebrafish

Quantitative investigation of optic nerve remyelination afte...

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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： Su-Qi ZOU Wu YIN Bing HU~* Laboratory of Neurodevelopment and Repair School of Life Sciences University of Science and Technology of China Hefei 230027 China

＜正＞Objective The dimension of the myelin sheath determines the insulating and conduction properties of the *** optic nerve crush,zebrafish retina ganglion cells could regenerate new axons to tectum and induce oligodendrocyte cells to wrap around *** axonal remyelination is very critical to visual function recovery during regeneration,we aim to study the relationship between myelination index and visual function in regenerationed optic nerve of adult *** We studied the dynamic change of axonal remyelination（myelination index） by using transmission electron microscopy（TEM）,and described the visual function with optokinetic response（OKR） at series timepoints after optic nerve *** 3～4 cm length adult zebrafish with normal visual function confirmed by optomotor response （OMR） were selected in this *** anesthetized,the optic nerve was crushed at 0.4～0.5 mm behind the eyeball for 10 sec with #5 *** different days after crush,OKR was applied to detect the visual *** were anesthetized in ice water,eyeballs were removed into 2.5%glutaraldehyde in 0.1 M（pH 7.4） phosphate buffer overnight at 4℃,washed three times with PB solution and fixed in 1%OsO4 for 1h,followed by standard sample preparations for electron microscopy *** There was an acute swelling of RGC axons at 3 h post crush（C3h）.The degenerated axons were cleared by microglia/macrophage around 7 days post crush（C7d）.The regenerationed axons were increased starting at C14d,and the myelination index in C87d was nearly eighty percent of normal *** zebrafish OKR,which was first established in our lab,was used to detect the visual function before electron microscopy of each *** results showed that visual function was completely disappeared at C3h,recovered at ClOd and reached half of normal level at *** OKR of 6 weeks post-injury was near to a platform at eighty percent of normal *** The regenerationed RGC axon

关键词： adult zebrafish optic nerve crush electron microscopy remyelination rate OKR

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Small extracellular vesicles derived from human induced pluripotent stem cell-differentiated neural progenitor cells mitigate retinal ganglion cell degeneration in a mouse model of optic nerve injury

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Neural Regeneration Research 2025年第2期20卷 587-597页

作者： Tong Li Hui-Min Xing Hai-Dong Qian Qiao Gao Sheng-Lan Xu Hua Ma Zai-Long Chi State Key Laboratory of Ophthalmology Optometry and Visual ScienceEye HospitalWenzhou Medical UniversityWenzhouZhejiang ProvinceChina National Clinical Research Center for Ocular Diseases Eye HospitalWenzhou Medical UniversityWenzhouZhejiang ProvinceChina

Several studies have found that transplantation of neural progenitor cells(NPCs)promotes the survival of injured ***,a poor integration rate and high risk of tumorigenicity after cell transplantation limits their clinical *** extracellular vesicles(sEVs)contain bioactive molecules for neuronal protection and *** studies have shown that stem/progenitor cell-derived sEVs can promote neuronal survival and recovery of neurological function in neurodegenerative eye diseases and other eye *** this study,we intravitreally transplanted sEVs derived from human induced pluripotent stem cells(hiPSCs)and hiPSCs-differentiated NPCs(hiPSC-NPC)in a mouse model of optic nerve *** results show that these intravitreally injected sEVs were ingested by retinal cells,especially those localized in the ganglion cell *** with hiPSC-NPC-derived sEVs mitigated optic nerve crush-induced retinal ganglion cell degeneration,and regulated the retinal microenvironment by inhibiting excessive activation of *** analysis further revealed that hiPSC-NPC derived sEVs transported neuroprotective and anti-inflammatory miRNA cargos to target cells,which had protective effects on RGCs after optic nerve *** findings suggest that sEVs derived from hiPSC-NPC are a promising cell-free therapeutic strategy for optic neuropathy.

关键词： exosome miRNA neural progenitor cell neurodegeneration neuroinflammation neuroprotection optic nerve crush optic neuropathy retinal ganglion cell small extracellular vesicles

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Mechanisms implicated in the contralateral effect in the central nervous system after unilateral injury:focus on the visual system

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Neural Regeneration Research 2021年第11期16卷 2125-2131页

作者： Fernando Lucas-Ruiz Caridad Galindo-Romero Virginia Albaladejo-García Manuel Vidal-Sanz Marta Agudo-Barriuso Departamento de Oftalmología Facultad de MedicinaUniversidad de Murcia and Instituto Murciano de Investigación Biosanitaria-Virgen de la Arrixaca(IMIBArrixaca)MurciaSpain

The retina,as part of the central nervous system is an ideal model to study the response of neurons to injury and disease and to test new *** the last decade is becoming clear that unilateral lesions in bilateral areas of the central nervous system trigger an inflammatory response in the contralateral uninjured *** effect has been better studied in the visual system where,as a rule,one retina is used as experimental and the other as *** retinas in unilateral models of retinal injury show neuronal degeneration and glial *** mechanisms by which this adverse response in the central nervous system occurs are discussed in this review,focusing primarily on the visual system.

关键词： bilateral effect brain glaucoma inflammation mirror effect optic nerve axotomy optic nerve crush retina spinal cord

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