大鼠组织中三磷酸肌醇受体亚型基因的表达  

Expression of Inositol Trisphosphate Receptor Subtypes mRNA in Rat Brain, Myocardium and Cultured Vascular Smooth Muscle Cells

作　　者：刘乃丰[1] 董莉[1] 何家声[1] 

机构地区：[1]南京铁道医学院附属医院心血管内科,南京210009

出　　处：《中国动脉硬化杂志》1999年第3期201-204,共4页Chinese Journal of Arteriosclerosis

基　　金：铁道部科技基金

摘　　要：为观察大鼠心肌、小脑和培养的主动脉血管平滑肌细胞中三磷酸肌醇受体亚型的基因表达，用异硫氰酸胍－ 酸酚氯仿一步法抽提组织总 Ｒ Ｎ Ａ，用特异性引物进行逆转录－ 聚合酶链式反应，β－ ａｃｔｉｎ 为内标半定量检测三磷酸肌醇受体亚型ｍ Ｒ Ｎ Ａ 的表达。将 Ｉ Ｉ型 Ｐ Ｃ Ｒ 产物重组、克隆并测序。发现三磷酸肌醇受体三个亚型在心肌、小脑和平滑肌细胞中均有表达，小脑组织中表达水平较高。 Ｐ Ｃ Ｒ 扩增片段分别为： Ⅰ型三磷酸肌醇受体５２５ ｂｐ （ 小脑） 或４０５ ｂｐ （ 心肌和平滑肌细胞） ； Ⅱ三磷酸肌醇受体４０５ ｂｐ ；Ⅲ型三磷酸肌醇受体１６９ ｂｐ 。序列分析发现克隆的Ⅱ三磷酸肌醇受体ｃ Ｄ Ｎ Ａ 序列与设计的目的ｃ Ｄ Ｎ Ａ一致，提示建立的逆转录－ 聚合酶链式反应方法能可靠地研究动物组织中三磷酸肌醇受体不同亚型的基因表达。Aim To establish a RT-PCR method for detecting expression of mRNA for inositol trisphosphate receptor (IP 3R) and to investigate expression of IP 3R subtypes in rat brain, myocardium and cultured vascular smooth muscle cells (VSMC). Methods Total RNA was isolated from different tissue of rat and was reversibly transcripted into cDNA. RT-PCR was performed with β-actin as internal label. Three oligonuclotide primers for each subtype of IP 3R were employed to amplify predicted DNA. Results It was found that all subtypes of IP 3R were expressed in cultured SMC. DNA sequencing indicated that amplified products of IP 3R Ⅱ subtype was identical to reported sequence from gene bank. Conclusion Our results suggested that RT-PCR established by us can be used as a reliable method for investigating expression of mRNA for IP 3R subtypes.

关 键 词：三磷酸肌醇 受体 基因表达 信号转导 平滑肌 

分 类 号：Q593.3[生物学—生物化学]