Filipeanu Catalin M, Zhou Fuguo, Wu Guangyu
Department of Pharmacology and Experimental Therapeutics, Louisiana State University Health Sciences Center, New Orleans, Louisiana, USA.
Methods Enzymol. 2008;438:217-26. doi: 10.1016/S0076-6879(07)38015-4.
Protein transport between intracellular organelles is coordinated by Rab GTPases. As an initial approach to defining the function of Rab GTPases in cardiomyocytes, our laboratory focused on Rab1, which regulates protein transport specifically from the endoplasmic reticulum (ER) to the Golgi apparatus. Our studies have demonstrated that adenovirus-driven expression of Rab1 promotes cell growth of primary cultures of neonatal cardiomyocytes in vitro and that transgenic expression of Rab1 in the myocardium induces cardiac hypertrophy in mouse hearts in vivo. These data provide strong evidence implicating that ER-to-Golgi protein transport functions as a regulatory site for control of cardiomyocyte growth. Here we describe a sets of methods used in our laboratory to characterize the function of Rab1 GTPase in modulating cardiac myocyte growth.
细胞内细胞器之间的蛋白质运输由Rab GTP酶协调。作为确定Rab GTP酶在心肌细胞中功能的初步方法,我们实验室重点研究了Rab1,它专门调节从内质网(ER)到高尔基体的蛋白质运输。我们的研究表明,腺病毒驱动的Rab1表达可促进体外新生心肌细胞原代培养物的细胞生长,并且Rab1在心肌中的转基因表达可在体内诱导小鼠心脏发生心肌肥大。这些数据提供了有力证据,表明内质网到高尔基体的蛋白质运输作为控制心肌细胞生长的调节位点发挥作用。在此,我们描述了我们实验室用于表征Rab1 GTP酶在调节心肌细胞生长中功能的一系列方法。