针对新生小鼠脑组织的两种病毒注射方法比较  

Comparison of two ways of virus injection in neonatal mouse brain tissue

作　　者：董禹豪 白占涛 魏忠诚 李袁梦 袁洁 张明 赵湘辉 DONG Yuhao;BAI Zhantao;WEI Zhongcheng;LI Yuanmeng;YUAN Jie;ZHANG Ming;ZHAO Xianghui(College of Life Sciences&Research Center for Resource Peptide Drugs,Shaanxi Engineering&Technological Research Center for Conversation&Utilization of Regional Biological Resources,Yan'an University,Yan'an 716099,China;Department of Neurobiology,School of Basic Medical Sciences,Air Force Medical University,Xi'an 710032,China;School of Basic Medicine,Shaanxi University of Chinese Medicine,Xianyang 712046,China)

机构地区：[1]延安大学生命科学学院多肽资源药物研究中心,陕西省区域生物资源保育与利用工程技术研究中心,陕西延安716099 [2]空军军医大学基础医学院神经生物学教研室,陕西西安710032 [3]陕西中医药大学基础医学院,陕西咸阳712046

出　　处：《空军军医大学学报》2024年第12期1370-1375,共6页Journal of Air Force Medical University

基　　金：国家自然科学基金面上项目(32371021,82071271);科技部科技创新2030-“脑科学与类脑研究”重大项目(2021ZD0201700)。

摘　　要：目的 选择侧脑室注射(LVI)与横窦注射(TSI)病毒的方法,比较不同途径介导的腺相关病毒感染新生小鼠脑组织的范围与脑区差异,为研究小鼠发育期特定脑区功能选择病毒注射方法提供参考。方法 对新生一日的C57BL/6仔鼠分别使用两种方法进行rAAV-GAD67-EGFP-WPREs病毒注射,并在第24日收集脑组织样品,制备冰冻切片观察病毒携带的荧光蛋白在小鼠脑内的感染表达范围。结果 LVI方法操作简单,但病毒表达的脑区范围较小,主要集中在大脑皮层。TSI方法操作步骤较复杂,病毒用量更大,但病毒感染的脑区范围更广。结论 不同注射方法会介导工具病毒在不同的脑区范围表达,其中LVI的方法更适合大脑皮层的相关发育研究,TSI的方法适合几乎全部脑区的发育研究。Objective To compare the differences in the range and brain regions between lateral ventricle injection(LVI)and transverse sinus injection(TSI)mediated adeno-associated virus expression in neonatal mice,and to provide reference for the selection of virus injection methods for the study of the function of specific brain regions in mouse development.Methods The newborn C57BL/6 mice were injected with rAAV-GAD67-EGFP-WPREs virus by LVI and TSI,respectively.Brain tissue samples were collected on the 24th day,and frozen brain slices were prepared to observe the expression range of virus-carried fluorescent protein in the mouse brain.Results The LVI method was easy to operate,but its range of virus injection was limited in the brain,mainly in the cerebral cortex.The TSI method was more complicated and required larger amount of virus,but more brain regions were infected under this method.Conclusion Different injection methods mediate the expression of virus in different brain regions,among which LVI is more suitable for the study of cerebral cortex development,and TSI is suitable for the developmental study of almost all brain regions.

关 键 词：侧脑室 横窦 腺相关病毒 脑发育 

分 类 号：R-332[医药卫生]