Study on expression of nesprins gene during mouse embryonic stem cells differentiation into cardiomyocytes in vitro

WANG Fang-jie; et al

doi:10.13406/j.cnki.cyxb.2010.05.012

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Study on expression of nesprins gene during mouse embryonic stem cells differentiation into cardiomyocytes in vitro

更新时间：2024-01-17

- Study on expression of nesprins gene during mouse embryonic stem cells differentiation into cardiomyocytes in vitro
- Journal of Chongqing Medical University Vol. 35, Issue 5, Pages: 652-656(2010)
- 作者机构：
  
  1. 重庆医科大学附属儿童医院心脏中心
  2. 剑桥大学心血管实验室
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2010.05.012
  CLC： R329
- Published：2010，
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[1]王芳洁,易岂建,郭鹏飞,刘晓燕,张秋萍,Shanahan C.nesprins基因在胚胎干细胞分化为心肌细胞过程中的表达[J].重庆医科大学学报,2010,35(05):652-656.

WANG Fang-jie, et al. Study on expression of nesprins gene during mouse embryonic stem cells differentiation into cardiomyocytes in vitro. [J]. Journal of Chongqing Medical University 35(5):652-656(2010)
[1]王芳洁,易岂建,郭鹏飞,刘晓燕,张秋萍,Shanahan C.nesprins基因在胚胎干细胞分化为心肌细胞过程中的表达[J].重庆医科大学学报,2010,35(05):652-656. DOI： 10.13406/j.cnki.cyxb.2010.05.012.

WANG Fang-jie, et al. Study on expression of nesprins gene during mouse embryonic stem cells differentiation into cardiomyocytes in vitro. [J]. Journal of Chongqing Medical University 35(5):652-656(2010) DOI： 10.13406/j.cnki.cyxb.2010.05.012.

摘要

目的:探讨nesprins基因在小鼠胚胎干细胞（Embryonic stem cell

ESC）分化为心肌细胞过程中的表达及作用。方法:体外培养小鼠ESC

采用5-氮杂胞苷为诱导剂诱导ESC分化为心肌细胞

并进行心肌细胞鉴定;RT-PCR检测分化过程中各时间点（d7、d7+3、d7+6、d7+9、d7+20）nesprin-1、nesprin-2基因mRNA的表达;免疫荧光检测各时间点nesprin-1蛋白的细胞定位。结果:nesprin-1、nesprin-2基因mRNA在ESC分化为心肌细胞过程中均有表达;nesprin-1基因mRNA在d7、d7+3、d7+6均表达较高

无显著性差异

随后表达呈下降趋势

在d7+20时表达最低（P<0.05）;nesprin-2基因mRNA在d7时表达最高

与d7+3、d7+6组间均有显著性差异（P<0.05）

且随着分化时间的延长表达逐渐下降

在d7+20时表达最低;nesprin-1蛋白在ESC分化过程中均有表达

定位于核被膜及核周间隙。结论:nesprin-1、nesprin-2基因表达高峰在生心细胞的诱导和特化时期

推测nesprins对维持心肌细胞发育

使ESC更完全的向心肌细胞分化起重要作用。

Abstract

Objective:To investigate expression and role of nesprins gene during mouse embryonic stem cell(ESC)differentiation into cardiomyocyte in vitro.Methods:Mouse ESC cultrued in combining 5-azacytidine as inducer with hanging drop cultures and the differentiated cells were identified with cardiac special genes and proteins;The mRNA expression of nesprin-1 and nesprin-2 genes were detected by reverse transcription-polymerase chain reaction(RT-PCR)

and sub-cellular location of nesprin-1 protein in nucleus was detected by immunofluorescence in d7

d7+3

d7+6

d7+9 and d7+20.Results:The expression of nesprin-1 and nesprin-2 genes mRNA can be found in the period of mouse ESC differentiation into cardiomyocyte.There is expression peak of nesprin-1 gene mRNA in d7

d7+3

d7+6

then declined gradually to the lowest in d7+20;expression peak of nesprin-2 gene mRNA in d7

then also declined gradually to the lowest in d7+20;The protein of nesprin-1 can be detected in the period of mouse ESC differentiation into cardiomyocyte

and mainly located in nuclear envelope and space around nuclei.Conclusion:The expression peak of nesprin-1 and nesprin-2 gene mRNA is interrelated with the phase of inducing and specialization of ESC differentiation into cardiac muscle cell

it can be speculated that nesprin genes has an important role in maintaining ESC differentiation into cardiac muscle cell.

关键词

nesprins胚胎干细胞心肌细胞分化小鼠

Keywords

NesprinsEmbryonic stem cellcardiomyocytedifferentiationmouse

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