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  

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 

机构地区：[1]Department of Ophthalmology,Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation,Sun Yat-sen Memorial Hospital,Sun Yat-sen University,Guangzhou,Guangdong Province,China [2]State Key Laboratory of Ophthalmology,Zhongshan Ophthalmic Center,Sun Yat-sen University,Guangzhou,Guangdong Province,China [3]Light Innovation Technology Ltd.,Hong Kong Special Administrative Region,China [4]Department of Ophthalmology&Visual Science,Shanghai Medical College,Fudan University,Shanghai,China [5]Department of Ophthalmology,Eye&ENT Hospital,Fudan University,Shanghai,China [6]Department of Ophthalmology,The First Affiliated Hospital of USTC,Division of Life Sciences and Medicine,University of Science and Technology of China,Hefei,Anhui Province,China

出　　处：《Neural Regeneration Research》2023年第4期913-921,共9页中国神经再生研究（英文版）

基　　金：supported by the National Natural Science Foundation of China,No.81870655(to MBY)。

摘　　要：Successful establishment of reconnection between retinal ganglion cells and retinorecipient regions in the brain is critical to optic nerve regeneration.However,morphological assessments of retinorecipient regions are limited by the opacity of brain tissue.In 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 injury.Specifically,we performed light-sheet imaging of whole brain tissue after a clearing process.We 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 brains.Unexpectedly,PRV724-traced neurons were more widely distributed compared with data from previous studies.We 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 nucleus.Our 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 injury.Thus,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 

分 类 号：R774[医药卫生—眼科]