宫内缺氧减缓小鼠普肯耶细胞发育和降低小脑肽的表达

doi:10.16098/j.issn.0529-1356.2021.03.003

解剖学报 ›› 2021, Vol. 52 ›› Issue (3): 344-351.doi: 10.16098/j.issn.0529-1356.2021.03.003

宫内缺氧减缓小鼠普肯耶细胞发育和降低小脑肽的表达

徐纪伟¹杨常青²陈小平³ 陈旭东^1*范文娟^1*

1.漯河医学高等专科学校组织学胚胎学教研室，河南漯河 462002; 2.漯河医学高等专科学校第三附属医院康复科，河南漯河 462002; 3.河南大学第一附属医院内分泌科，河南开封 475000

收稿日期:2019-11-29 修回日期:2020-01-09 出版日期:2021-06-06 发布日期:2021-06-06
通讯作者: 陈旭东;范文娟 E-mail:243538592@qq.com
基金资助:
河南省科技厅科技攻关项目;漯河医专博士启动基金

Intrauterine hypoxia slowing the development of Purkinje cells and reducing the expression of cerebellin in mice

XU Ji-wei¹YANG Chang-qing² CHEN Xiao-ping³ CHEN Xu-dong^1* FAN Wen-juan^1*

1.Department of Histology and Embryology, Luohe Medical College, He’nan Luohe 462002,China; 2.Department of Rehabilitation, the Third Affiliated Hospital of the Luohe Medical College, He’nan Luohe 462002,China;3.Department of Endocrine, the First Affiliated Hospital of He’nan University, He’nan Kaifeng 475000, China

Received:2019-11-29 Revised:2020-01-09 Online:2021-06-06 Published:2021-06-06
Contact: CHEN Xu-dong;FAN Wen-juan E-mail:243538592@qq.com

摘要/Abstract

摘要：

目的探讨宫内缺氧对出生后小鼠普肯耶细胞的发育及小脑肽表达的影响。方法 20只健康成年昆明小鼠孕鼠随机分为2组：对照组与缺氧组，每组10只。自母鼠孕14 d时开始，将缺氧组的母鼠置入动物缺氧箱内，制作宫内缺氧动物模型。待母鼠分娩后，实验动物分为缺氧组和对照组，每组内有出生当日（P0）、P5、P9、P14、P21和P30这 6个年龄段，每年龄段小鼠各取5只。取小脑组织做振荡切片，利用免疫荧光技术检测钙结合蛋白（calbindin）阳性普肯耶细胞的发育变化，利用小脑肽1(CBLN1)、CBLN4与calbindin双重标记的方法检测普肯耶细胞突起上小脑肽的表达。利用Western blotting对各组各时间点动物小脑组织内小脑肽蛋白表达进行半定量分析。结果与同龄对照组相比，缺氧组小脑普肯耶细胞数量减少，树突分支减少且排列混乱，皮质内CBLN1与CBLN4的表达明显降低。Western blotting结果与免疫荧光结果吻合。结论宫内缺氧导致小脑普肯耶细胞发育异常以及突触改变。

关键词: 缺氧, 小脑, 普肯耶细胞, 小脑肽, 免疫印迹法, 小鼠

Abstract:

Objective To investigate the effects of the development of Purkinje cells and expression of cerebellin in postnatal mice with intrauterine hypoxia. Methods Twenty healthyice were randomly divided into two groups: control group and hypoxia group, with 10 mice in each group. Mice in the hypoxia group were placed in the hypoxia chamber of the animal since the 14th day of gestation to make an animal model of intrauterine hypoxia. After the mother gave birth, the experimental animals were divided into hypoxia group and control group. There were 6 age groups including postnatal day (P)0, P5, P9, P14, P21 and P30 in each group, and 5 mice in each age group. The cerebellum tissue was taken for vibrating sectioning. The developmental changes of calbindin-positive Purkinje cells were detected by immunofluorescence technique. The expression of cerebellin in Purkinje cell protuberances was detected by cerebellin (CBLN)1, CBLN4 and calbindin double labeling. Finally, Western blotting was used to semiquantitatively analyze the protein expression of cerebellar peptide in cerebellum at each time point. Results Compared with the control group of the same age, the number of cerebellar Purkinje cells in the hypoxic group decreased, the dendritic branches decreased, and the arrangement was disordered, and the expression of CBLN1 and CBLN4 in the cortex were significantly reduced. Conclusion Intrauterine hypoxia leads to abnormal development of the cerebellar Purkinje cells and synaptic changes.

Key words: Hypoxia, Cerebellum, Purkinje cell, Cerebellin, Western blotting, Mouse

中图分类号:

R329

徐纪伟杨常青陈小平陈旭东范文娟. 宫内缺氧减缓小鼠普肯耶细胞发育和降低小脑肽的表达[J]. 解剖学报, 2021, 52(3): 344-351.

XU Ji-wei YANG Chang-qing CHEN Xiao-ping CHEN Xu-dong FAN Wen-juan. Intrauterine hypoxia slowing the development of Purkinje cells and reducing the expression of cerebellin in mice[J]. Acta Anatomica Sinica, 2021, 52(3): 344-351.

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宫内缺氧减缓小鼠普肯耶细胞发育和降低小脑肽的表达

Intrauterine hypoxia slowing the development of Purkinje cells and reducing the expression of cerebellin in mice

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