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浙江大学学报（医学版）
浙江大学学报（医学版）  2012, Vol. 41 Issue (4): 366-372    DOI: 10.3785/j.issn.1008-9292.2012.04.003
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代谢型谷氨酸受体4在ES细胞体外分化为心肌细胞中的表达研究
郑蓓1，周立民1，赵青威2，张幸国2，朱丹雁1
1.浙江大学药学院 药理毒理与生化药学研究所，浙江 杭州 310058； 2.浙江大学医学院附属第一医院，浙江 杭州 310003
Expression of metabotropic glutamate receptor 4 in cardiomyocytes differentiated from mouse embryonic stem cells in vitro
ZHENG Bei1, ZHOU Li-min1, ZHAO Qing-wei 2, ZHANG Xing-guo 2, ZHU Dan-yan1
1.Institute of Pharmacology, Toxicology, and Biochemical Pharmaceutics, Zhejiang University, Hangzhou 310058, China; 2.The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China
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摘要： 目的：探索代谢型谷氨酸受体4（metabotropic glutamate receptor 4，mGluR4）在小鼠胚胎干细胞（embryonic stem cells，ES细胞）体外分化为心肌细胞中的表达情况。
方法：采用悬滴培养法，体外诱导ES细胞分化为心肌细胞，分别以维甲酸（retinoic acid，RA）和溶剂二甲亚砜（dimethyl sulfoxide，DMSO）作为阳性对照和阴性对照；利用免疫荧光法和流式细胞术，鉴定ES细胞衍生心肌细胞团中心肌特异性蛋白α-actinin/Troponin-T和mGluR4共表达情况；用Western Blot法，考察ES细胞定向分化为心肌细胞过程中mGluR4蛋白表达水平；同时观察了小鼠胚胎心脏发育过程中mGluR4基因和蛋白表达变化。
结果：ES细胞及其分化来的早期心肌细胞团均表达mGluR4，随着心肌细胞逐渐成熟，mGluR4蛋白表达水平下调。流式细胞术结果显示，贴壁分化11 d的细胞团中mGluR4与心肌特异性蛋白Troponin-T共表达细胞比率仅为（3.00±1.00）%。同时，小鼠胚胎心脏发育过程中mGluR4基因和蛋白表达也呈下降趋势，直至成体小鼠心脏中不表达mGluR4蛋白。
结论：ES细胞体外分化为心肌细胞过程中mGluR4有表达并呈下调趋势，与小鼠体内心脏发育过程中mGluR4表达情况基本一致。
关键词： 代谢型谷氨酸受体4； 胚胎干细胞； 肌细胞;  心脏/细胞学； 受体;  谷氨酸； 心肌细胞    
Abstract: Objective： To investigate the expression of metabotropic glutamate receptor 4 (mGluR4) in cardiomyocytes differentiated from mouse embryonic stem cells (ES cells).
Methods： ES cells were differentiated into cardiomyocytes with hanging-drop cultures. Retinoic acid (RA) and dimethyl sulfoxide (DMSO) were used as positive and negative controls, respectively. The co-expression of cardiac sarcomeric protein (α-actinin or troponin-T) and mGluR4 were verified by immunocytochemistry and flow cytometry analysis. The mRNA and protein expressions of mGluR4 were verified by RT-PCR and Western blot analysis, respectively. Meanwhile, the expression of mGluR4 in prenatal mouse heart was also examined.
Results： mGluR4 was expressed in both mouse ES cells and ES cell-derived cardiomyocytes. The level of mGluR4 protein expression decreased during the maturation of the cardiomyocytes. The co-expression rate of mGluR4 and Troponin T in the beating embryoid bodies (EBs) was only（3.00±1.00）%. On the other hand, mGluR4 gene and protein expressions showed remarkable down-regulation in the development of mouse fetal heart, which was not detected in mouse adult heart.
Conclusion： The expression of mGluR4 is down-regulated in the cardiomyocyte differentiation of ES cells. The trend of expression is consistent with that in the prenatal mouse heart development.
Key words: Metabotropic glutamate receptor 4    Embryonic stem cells    Myocytes,cardiac/cytol    Receptors,glutamate    Cardiomyocytes
出版日期: 2012-07-25
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郑蓓;周立民;赵青威;张幸国;朱丹雁. 代谢型谷氨酸受体4在ES细胞体外分化为心肌细胞中的表达研究[J]. 浙江大学学报（医学版）, 2012, 41(4): 366-372.

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https://www.zjujournals.com/med/CN/Y2012/V41/I4/366

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