小脑发育过程中Atoh1时空表达模式

doi:10.16098/j.issn.0529-1356.2023.02.002

解剖学报 ›› 2023, Vol. 54 ›› Issue (2): 134-141.doi: 10.16098/j.issn.0529-1356.2023.02.002

小脑发育过程中Atoh1时空表达模式

薛赛赛¹ 陈丽萍² 李莹³ 吴燕² 吴海涛^1,2*

1.青岛大学基础医学院生理学与病理生理学系，青岛 266071; 2.军事科学院军事医学研究院军事认知与脑科学研究所，北京 100850; 3.首都医科大学基础医学院神经生物学系，北京 100069

收稿日期:2022-09-09 修回日期:2022-11-02 出版日期:2023-04-06 发布日期:2023-04-06
通讯作者: 吴海涛 E-mail:wuht@bmi.ac.cn
基金资助:
国家自然科学基金;国家自然科学基金;科技部重点研发专项;科技创新2030-“脑科学与类脑研究”重大项目

Spatial and temporal expression patterns of Atoh1 during mice cerebellar development

XUE Sai-sai¹ CHEN Li-ping² LI Ying³ WU Yan² WU Hai-tao^1,2*

1.Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Qingdao University, Qingdao 266071, China;

2.Institute of Military Cognition and Brain Sciences, Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 100850, China; 3.Department of Neurobiology,School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China

Received:2022-09-09 Revised:2022-11-02 Online:2023-04-06 Published:2023-04-06
Contact: WU Hai-tao E-mail:wuht@bmi.ac.cn

摘要/Abstract

摘要：

目的探究小脑发育过程中转录因子Atoh1 mRNA及其蛋白的时空表达模式。方法取不同发育阶段的小鼠小脑冷冻切片，RNA scope技术分析Atoh1 mRNA在小脑中的时空表达模式，每组 n=3。同时，使用Atoh1转基因和基因敲入两种不同遗传学策略的报告小鼠，采用免疫荧光染色技术分析Atoh1在蛋白水平的时空表达，每组n=3。结果 Atoh1 mRNA在胚胎小脑菱唇和外颗粒细胞层中呈高表达；Atoh1-3 ^*V5-P2A-tdTomato/+基因敲入小鼠染色结果显示，Atoh1主要表达在菱唇和有增殖能力的外颗粒细胞层的外层；Atoh1-Cre;tdTomato/+转基因小鼠显示，Atoh1阳性细胞在菱唇和整个颗粒细胞发育过程中包括外颗粒细胞层和内颗粒细胞层中均呈高表达。结论小脑发育过程中，Atoh1 mRNA和蛋白在胚胎期的菱唇和出生后的外颗粒细胞层中高丰度表达；但在蛋白水平上，基因敲入小鼠和转基因小鼠Atoh1在菱唇和外颗粒细胞层中的表达略有不同。

关键词: 小脑, 发育, Atoh1, 颗粒前体细胞, RNAscope, 小鼠

Abstract:

Objective To investigated the spatial and temporal expression of Atoh1 mRNA and protein level during the cerebellar development. Methods The frozen cerebellum sections of mice at different developmental stages were used to analyze the temporal and spatial expression patterns of Atoh1 mRNA by RNA scope technique, n=3 for each group. Meanwhile, two reporter mice with different genetic modification method were utilized to analyze the spatialand temporal expression of Atoh1 at the protein level by immunofluorescence staining, n=3 for each group. Results Atoh1 mRNA was highly expressed in the embryonic cerebellar rhombic lip (RL) and the external granule layer (EGL). At the protein level, staining result of Atoh1-3*V5-P2A-tdTomato/+ knockin mice showed that Atoh1 was mainly expressed in the RL and the outer layer of the proliferative EGL. Atoh1-Cre;tdTomato/+ transgenic reporter mice showed that tdTomato positive cells were highly expressed in the EGL and the inner granule cell layer (IGL) during the whole granule cells development. Conclusion During cerebellar development, Atoh1 mRNA and protein were highly expressed in the embryonic rhombic lip and postnatal external granule layer. However, at the protein level, the expression of Atoh1 in the rhombic lip and external granule layer were different between knockin mice and transgenic mice.

Key words: Cerebellum, Development, Atoh1, Granule neuron precursor cell, RNA scope, Mouse

中图分类号:

薛赛赛陈丽萍李莹吴燕吴海涛 . 小脑发育过程中Atoh1时空表达模式[J]. 解剖学报, 2023, 54(2): 134-141.

XUE Sai-sai CHEN Li-ping LI Ying WU Yan WU Hai-tao. Spatial and temporal expression patterns of Atoh1 during mice cerebellar development[J]. Acta Anatomica Sinica, 2023, 54(2): 134-141.

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小脑发育过程中Atoh1时空表达模式

Spatial and temporal expression patterns of Atoh1 during mice cerebellar development

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