[1]陈 文,温明达,董大川,等.PP2A Cα敲除小鼠心肌细胞模型的构建[J].南京师大学报(自然科学版),2013,36(02):74-77.
　Chen Wen,Wen Mingda,Dong Dachuan,et al.Generation of PP2A Cα Knockout Model in Neonatal Mouse Ventricular Myocytes[J].Journal of Nanjing Normal University(Natural Science Edition),2013,36(02):74-77.
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PP2A Cα敲除小鼠心肌细胞模型的构建()
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《南京师大学报（自然科学版）》[ISSN:1001-4616/CN:32-1239/N]

卷:
第36卷
期数:
2013年02期
页码:
74-77
栏目:
生命科学
出版日期:
2013-06-30

文章信息/Info

Title:
Generation of PP2A Cα Knockout Model in Neonatal Mouse Ventricular Myocytes
文章编号:
1001-4616(2013)02-0074-04
作者:
陈 文; 温明达; 董大川; 高艳红; 张 朝
南京师范大学生命科学学院,江苏省分子医学生物技术重点实验室,江苏 南京 210023
Author(s):
Chen Wen; Wen Mingda; Dong Dachuan; Gao Yanhong; Zhang Zhao
Jiangsu Key Laboratory for Molecular and Medical Biotechnology,School of Life Sciences,Nanjing Normal University,Nanjing 210023,China
关键词:
PP2A Cα; 新生小鼠心肌细胞; 带重组酶Cre的腺病毒
Keywords:
PP2A Cα; neonatal mouse ventricular myocytes; Cre-adenovirus
分类号:
Q2-33
文献标志码:
A
摘要:
体外构建PP2A Cα敲除的原代小鼠心室肌细胞模型.选择性地将雄性PP2A Cαfl/fl,Cre/-小鼠与雌性PP2A Cαfl/fl,Cre/-小鼠交配.新生小鼠心脏经胰酶消化后差速贴壁获得原代小鼠心室肌细胞,用带有Cre的腺病毒侵染心肌细胞,经荧光显微镜观察和Western-blot检测,分析PP2A Cα的敲除效果.将基因型为PP2A Cαfl/fl,Cre/-的雌雄小鼠进行交配,得到其同样基因型的后代,进而可获得足量基因型为PP2A Cαfl/fl,Cre/-的原代小鼠心肌细胞.用带有重组酶Cre的腺病毒侵染细胞48 h后,可见带有绿色荧光的心肌细胞; Western-blot检测显示,PP2A Cα在Cre腺病毒侵染的心肌细胞可下调60%～80%.本研究成功建立了PP2A Cα敲除的原代小鼠心肌细胞模型.
Abstract:
To generate PP2A Cα knockout model in neonatal mouse ventricular myocytes.Male PP2A Cαfl/fl,Cre/- were selectively mated with female PP2A Cαfl/fl,Cre/- mice to obtain abundant offsprings with the same genotype.The neonatal mouse ventricular myocytes were digested by trypsinization and subsequently infected with Cre-adenovirus.The effects of adenovirus infection were observed by fluorescence microscope and identified by western blot analysis.The offsprings in aboundance were obtained by seting up a mating between male and female of PP2A Cαfl/fl,Cre/-mice,as a result,an abundance of neonatal mouse ventricular myocytes with the genotype of PP2A Cαfl/fl,Cre/-were available for Ad-Cre infection.Green fluorescence was observed after GFP labeled Cre-adenovirus(Cre-Ad)infection for 48 h.The expression level of PP2A Cα protein in Cre-Ad infected myocytes was 60%～80% lower than control.The PP2A Cα knockout model in neonatal mouse ventricular myocytes was successfully generated.

参考文献/References:

[1] Janssens V,Goris J.Protein phosphatase 2A:a highly regulated family of serine/threonine phosphatases implicated in cell growth and signalling[J].Biochem J,2001,353(Pt 3):417-439.
[2]Shi Y.Serine/threonine phosphatases:mechanism through structure[J].Cell,2009,139(3):468-484.
[3]Gu P.Generation of Ppp2Ca and Ppp2Cb conditional null alleles in mouse[J].Genesis,2012,50(5):429-436.
[4]Neumann J.Increased expression of cardiac phosphatases in patients with end-stage heart failure[J].J Mol Cell Cardiol,1997,29(1):265-272.
[5]Shi J.Protein phosphatase 2A effectively modulates basal L-type Ca(2+)current by dephosphorylating Ca(v)1.2 at serine 1866 in mouse cardiac myocytes[J].Biochem Biophys Res Commun,2012,418(4):792-798.

备注/Memo

备注/Memo:
收稿日期:2013-01-26.
基金项目:973课题(2006CB943500)子项目、国家自然基金课题(31171302)、江苏高校优势学科建设工程项目(164320H106).
通讯联系人:张朝,教授,博士,研究方向:心肌离子通道功能及其调控.E-mail:zhangzhao@njnu.edu.cn
更新日期/Last Update: 2013-06-30