Ifitm3敲除抑制小鼠神经干细胞增殖和分化  

Ifitm3 knockout inhibits the proliferation and differentiation of neural stem cells in mice

作　　者：王凯瑜 雷雪裴 黄艺滢 石桂英 乐涵薇 王杰 林羿凡 汤家鸣 白琳 WANG Kaiyu;LEI Xuepei;HUANG Yiying;SHI Guiying;YUE Hanwei;WANG Jie;LIN Yifan;TANG Jiaming;BAI Lin(Institute of Laboratory Animal Sciences,CAMS&PUMC,NHC Key Laboratory of Human Disease Comparative Medicine,National Human Diseases Animal Model Resource Center,Beijing 100021,China)

机构地区：[1]中国医学科学院医学实验动物研究所,国家卫生健康委员会人类疾病比较医学重点实验室,国家人类疾病动物模型资源库,北京100021

出　　处：《中国实验动物学报》2024年第6期691-701,共11页Acta Laboratorium Animalis Scientia Sinica

基　　金：中国医学科学院创新项目(2021-I2M-1-035);国家重点研发计划(2022YFA1103803)

摘　　要：目的建立干扰素诱导跨膜蛋白3基因(Ifitm3)敲除小鼠模型,探究Ifitm3对小鼠神经干细胞增殖和分化的影响。方法利用CRISPR/Cas9技术,建立Ifitm3敲除小鼠模型,通过基因型鉴定和Western Blot检测IFITM3的敲除效果;利用苏木素-伊红(HE)染色和流式细胞术等方法分析Ifitm3敲除小鼠和野生型小鼠的表型差异。分离并培养野生型和Ifitm3敲除小鼠成体神经干细胞,统计神经球数量和大小,qRT-PCR、Western Blot、免疫荧光技术检测神经干细胞增殖和分化的能力。结果成功建立Ifitm3敲除小鼠模型,Ifitm3敲除小鼠发育正常,组织病理学及免疫系统未见明显异常。体外实验显示Ifitm3敲除抑制小鼠神经干细胞的自我更新潜能,导致神经干细胞增殖能力下降,并且抑制神经干细胞向未成熟神经元和星形胶质细胞分化。结论IFITM3缺失导致神经干细胞增殖和分化能力下降,IFITM3可能参与了神经干细胞的功能调节。Objective To establish interferon⁃induced transmembrane protein 3(Ifitm3)knockout mice and to explore the effects of Ifitm3 on the proliferation and differentiation of adult neural stem cells of mice(aNSCs).Methods IFITM3 knockout mice were established by the CRISPR/Cas9 method and identified by genotype identification and Western Blot.The differences between Ifitm3⁃knockout mice and wild⁃type mice were analyzed by hematoxylin⁃eosin(HE)staining and flow cytometry.The aNSCs of wild⁃type mice and Ifitm3⁃knockout mice were isolated and cultured,the number and size of neurospheres were detected,The ability of aNSCs to proliferate and differentiate were detected by quantitative reverse⁃transcription polymerase chain reaction,Western Blot,and immunofluorescence.Results Ifitm3⁃knockout mice were successfully established.The mice developed normally,and there were no obvious abnormalities either histopathologically or the immune system.In vitro experiments showed that Ifitm3 knockout inhibited the self⁃renewal potential of aNSCs,led to a decrease in the proliferation ability of aNSCs,and inhibited the differentiation of aNSCs into immature neurons and astrocytes.Conclusions This study finds that a lack of IFITM3 result in the ability of aNSCs to proliferate and differentiate decreased,IFITM3 may regulate the function of aNSCs.

关 键 词：干扰素诱导跨膜蛋白3 基因敲除 神经干细胞 增殖 神经分化 

分 类 号：Q95-33[生物学—动物学]