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 杨常青 陈小平 陈旭东 范文娟

解剖学报 ›› 2021, Vol. 52 ›› Issue (3) : 344-351.

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解剖学报 ›› 2021, Vol. 52 ›› Issue (3) : 344-351. DOI: 10.16098/j.issn.0529-1356.2021.03.003
神经生物学

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*
作者信息 +

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

  • XU Ji-wei1 YANG Chang-qing2 CHEN Xiao-ping3 CHEN Xu-dong1* FAN Wen-juan1*
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文章历史 +

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。

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.

关键词

缺氧 / 小脑 / 普肯耶细胞 / 小脑肽 / 免疫印迹法 / 小鼠

Key words

Hypoxia / Cerebellum / Purkinje cell / Cerebellin / Western blotting / Mouse

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导出引用
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 https://doi.org/10.16098/j.issn.0529-1356.2021.03.003
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 https://doi.org/10.16098/j.issn.0529-1356.2021.03.003
中图分类号: R329    

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基金

河南省科技厅科技攻关项目;漯河医专博士启动基金

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