Combining CUBIC Optical Clearing and Thy1-YFP-16 Mice to Observe Morphological Axon Changes During Wallerian Degeneration  

作　　者：Yu-song YUAN Fei YU Su-ping NIU Hao LU Yu-hui KOU Hai-lin XU 

机构地区：[1]Department of Trauma and Orthopaedics,Peking University People's Hospital,Peking University',Beijing 100044,China [2]National and Local Joint Engineering Research Center of Orthopaedic Biomaterials,Department of Bone&Joint Surgery,Peking University Shenzhen Hospital,Shenzhen 518036,China [3]Key Laboratory of Trauma and Neural Regeneration(Peking University),Ministry of Education,Beijing 100044,China [4]Office of Academic Research,Peking University People's Hospital,Peking University,Beijing 100044,China [5]Diabetic Foot Treatment Center,Peking University People's Hospital,Peking University',Beijing 100044,China

出　　处：《Current Medical Science》2021年第5期944-952,共9页当代医学科学（英文）

基　　金：supported by grants from the National Key Research and Development Program of China(No.2016YFC1101604);the Fundamental Research Funds for the Central Universities;Clinical Medicine Plus X-Young Scholars Project of Peking University China(No.PKU2020LCXQ020);Guangdong Basic and Applied Basic Research Foundation(No.2019A1515110983,No.2021A1515012586);Bethune Charitable Foundation and CSPC Osteoporosis Research Foundation Project(No.G-X-2020-1107-21).

摘　　要：Objective:Wallerian degeneration is a pathological process closely related to peripheral nerve regeneration following injury,and includes the disintegration and phagocytosis of peripheral nervous system cells.Traditionally,morphological changes are observed by performing immunofluorescence staining after sectioning,which results in the loss of some histological information.The purpose of this study was to explore a new,nondestmetive,and systematic method for observing axonal histological changes during Wallerian degeneration.Methods:Thirty male Thy1-YFP-16 mice(SPF grade,6 weeks old,20±5 g)were randomly selected and divided into clear,unobstructed brain imaging cocktails and computational analysis(CUBIC)optical clearing(n=15)and traditional method groups(n=15).Five mice in each group were sacrificed at 1st,3rd,and 5th day following a crush operation.The histological axon changes were observed by CUBIC light optical clearing treatment,direct tissue section imaging,and HE staining.Results:The results revealed that,compared with traditional imaging methods,there was no physical damage to the samples,which allowed for three-dimensional and deep-seated tissue imaging through CUBIC.Local image information could be nicely obtained by direct fluorescence imaging and HE staining,but it was difficult to obtain image information of the entire sample.At the same time,the image information obtained by fluorescence imaging and HE staining was partially lost.Conclusion:The combining of CUBIC and Thy1-YFP transgenic mice allowed for a clear and comprehensive observation of histological changes of axons in Wallerian degeneration.

关 键 词：Wallerian degeneration AXON optical clearing clear unobstructed brain imaging cocktails and computational analysis Thy1-YFP transgenic mice 

分 类 号：R73[医药卫生—肿瘤]