低压低氧刺激后小鼠小脑中RNA m6A甲基化调控因子表达的变化  

Expression changes of RNA m6A regulators in mouse cerebellum affected by hypobaric hypoxia stimulation

作　　者：肖灵凤 马春卉 赵顺莉 李晴 刘春英[1] 牛亚梅[1] 佟伟民[1] Xiao Lingfeng;Ma Chunhui;Zhao Shunli;Li Qing;Liu Chunying;Niu Yamei;Tong Weimin(Department of Pathology,Institute of Basic Medical Science Chinese Academy of Medical Science,School of Basic Medicine Peking Union Medical College,Beijing 100005,China)

机构地区：[1]中国医学科学院基础医学研究所暨北京协和医学院基础学院病理学系,北京100005

出　　处：《中华病理学杂志》2024年第5期452-457,共6页Chinese Journal of Pathology

基　　金：中央高校基本科研业务费专项资金资助(3332022037)。

摘　　要：目的探讨RNA m6A甲基化在低压低氧引起的小脑发育障碍中可能发挥的作用。方法将出生后5 d的C57/BL6小鼠放置于低压低氧饲养舱中连续处理9 d后,首先通过体重、脑重以及HE染色的方式来初步分析小鼠小脑发育情况;其次,利用免疫组织化学和免疫荧光染色法检测小脑中不同类型细胞特异性标志物的表达以解析其细胞结构的紊乱程度;然后利用即时荧光定量聚合酶链反应、Western blot和免疫组织化学染色方法检测小鼠小脑中RNA m6A主要调控因子的表达变化。结果相比于对照组小鼠,低压低氧小鼠体重、脑重以及小脑体积显著减小(P<0.01);不同类型细胞特异性标志物的免疫染色结果显示,NeuN(成熟神经元)表达下降,Calbindin-D28K(浦肯野细胞)和胶质纤维酸性蛋白(星形胶质细胞)的表达下降(P<0.01),且细胞分布紊乱;RNA m6A主要调控因子的表达分析结果显示,甲基转移酶METTL3在低压低氧小鼠小脑中的表达显著下调(P<0.05)。结论低压低氧刺激会造成小鼠小脑发育障碍,成熟神经元、浦肯野细胞和星形胶质细胞结构异常;与常压常氧小鼠相比,低压低氧刺激后小鼠小脑METTL3表达下降,提示其介导的RNA m6A甲基化可能在低压低氧引起的小脑发育障碍过程中发挥重要的作用。Objective To investigate the role of RNA m6A methylation in mediating cerebellar dysplasia through analyzing the phenotypes of the mouse cerebella and the expression of several key m6A regulators upon hypobaric hypoxia treatment.Methods Five-day old C57/BL6 mice were exposed to hypobaric hypoxia for 9 days.The status of mouse cerebellar development was analyzed by comparing the body weights,brain weights and histological features.Immunostaining of cell-type-specific markers was performed to analyze the cerebellar morphology.Real-time PCR,Western blot and immunohistochemical staining were performed to detect the expression of key m6A regulators in the mouse cerebella.Results Compared with the control,the body weights,brain weights and cerebellar volumes of hypobaric hypoxic mice were significantly reduced(P<0.01).The expression of specific markers in different cells,including NeuN(mature neuron),Calbindin-D28K(Purkinje cell)and GFAP(astrocyte),was decreased in hypobaric hypoxic mouse cerebella(P<0.01),accompanied with disorganized cellular structure.The expression of methyltransferase METTL3 was significantly down-regulated in the cerebella of hypobaric hypoxic mice(P<0.05).Conclusions Hypobaric hypoxia stimulation causes mouse cerebellar dysplasia,with structural abnormalities in mature granular neurons,Purkinje cells and astrocytes.Expression of METTL3 is decreased in hypobaric hypoxic mice cerebellum compared with that of normobaric normoxic mice,suggesting that its mediated RNA m6A methylation may play an important role in hypobaric hypoxia-induced mouse cerebellar dysplasia.

关 键 词：小脑 细胞低氧 RNA甲基化 甲基转移酶类 

分 类 号：R742.82[医药卫生—神经病学与精神病学]