|本期目录/Table of Contents|

[1]刘丹妮,高小方,姚成立,等.大鼠心室肌HCN4质粒在HEK293细胞的表达及电流特征[J].宁夏医科大学学报,2008,(02):146-149.
 LIU Dan ni,LI Xiao ying,GAO Xiao fang,et al.Current Characteristics and Expression of HCN4 Myocytes in Rat Ventricular Myocytes into HEK293[J].Ningxia Medical University,2008,(02):146-149.
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大鼠心室肌HCN4质粒在HEK293细胞的表达及电流特征(PDF)
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《宁夏医科大学学报》[ISSN:1005-8486/CN:64-1029/R]

卷:
期数:
2008年02期
页码:
146-149
栏目:
论著
出版日期:
2008-03-20

文章信息/Info

Title:
Current Characteristics and Expression of HCN4 Myocytes in  Rat Ventricular Myocytes into HEK293
作者:
刘丹妮1高小方1姚成立2李泱3
1宁夏回族自治区人民医院,银川750021;2银川市第一人民医院,银川750001;  3解放军总医院老年心血管病研究所,北京100853
Author(s):
LIU Danni LI Xiaoying GAO Xiaofang LIU Ying YAO Chenli LI Yang
Dept of Cardiology, Ningxia People's Hospital,Yinchuan 750021
关键词:
超极化激活环核苷酸门控阳离子通道异源性表达膜片钳 大鼠
Keywords:
hyperpolarization activated cyclic nucleotide gated catio n channelheterologously expressedpatch clamp recordingsrat
分类号:
R33-33
DOI:
-
文献标志码:
A
摘要:
目的 研究大鼠心室肌细胞超极化环核苷酸门控阳离子通道(HCN4)基因瞬时转染人 胚胎肾细胞(HEK293)表达和通道电流特征。方法 利用脂质体介导大鼠HC N4基因pTracer-C MV-GFP穿梭载体,转染HEK293细胞进行扩增,用逆转录-聚合酶链反应(RT- PCR)检测HCN 4 mRNA表达,采用全细胞膜片钳技术记录HEK293细胞上HCN4电流。结果 大鼠起搏通道基因 瞬时转染HEK293细胞,记录到一系列超极化内向离子流,该电流呈电压、时间依赖性,没有 明显的失活,在-150 mV时电流幅值为(540±241)pA,电流密度为(96±07)pA/pF (n =12),半激活电压(V1/2)为(-1057±120)mV,稳态激活曲线斜率(k)为-15 7mV,H CN4电流翻转电位为(-840±79)mV。结论 应用脂质体转染方法成功 进行了起搏通道HCN4基因的异源性表达。
Abstract:
Objective To explore the current characteristics and expressi on of H CN4 myocytes in rat ventricular myocytes into HEK293 Methods Th e HCN4 gene fra gment was cloned into the shuttle plasmid pTracerCMVGFP to form the transfer v ectorThe mRNA expression was detected by reverse transcriptasePCR method aft er being transfected into the HEK293 cell using Lipofactamine2000 reagent for ampl ification,then to use whole cell patch clamp recording the current Resu lts[WTB1]An inward current was recorded in response to hyperpolarization activaton in trans fected HEK293 cell The mHCN4 channel began to activate in voltage and time depen dent manner without deactivation threshold voltage and potential of halfmaxima l activation(V1/2)was -70mV,(-1057±120)mV ,respectively. [WT5H 1]Conclusion HCN4 g ene is successfly heterologously expressed by the method of Lipofactamine transf ected

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期:2007-11-13 基金项目:国家自然科学基金项目(30570739) 作者简介:刘丹妮(1973-),女,在读硕士研究生,主治医师,主要从事心血管电 生理的研究。E-mail:liudanni216@sina.com.cn 通信作者:李泱,男,研究员,研究生导师,主要从事心脏基础电生理研究。 E-mail:liyangbsh@163.com.cn
更新日期/Last Update: 2008-03-20