Marcil Valérie, Seidman Ernest, Sinnett Daniel, Sanchez Rocio, Spahis Schohraya, Sané Alain, Levy Emile
Research Institute, McGill University, Campus MGH, C10.148.6, Montreal, Quebec, Canada.
Cell Physiol Biochem. 2014;33(6):1815-26. doi: 10.1159/000362960. Epub 2014 Jun 17.
BACKGROUND/AIMS: Sar1b GTPase (Sar1b) represents an obligatory component of COPII vesicles that bud from the endoplasmic reticulum to transport proteins to the Golgi apparatus. Its genetic mutations lead to a severe disorder known as chylomicron retention disease. Despite growing knowledge on Sar1b, little is known about it tissue distribution and regulation, which constitute the aims of the present study. We aimed to determine the Sar1b tissue distribution and modulation by a high-fat diet and gene forcing using transgenic mice in comparison to wild-type mice. Methods : The expression pattern of Sar1b was studied in different organs of wild-type and Sar1b transgenic mice by qRT-PCR and Western blot. The effect of transgenesis and insulin resistance induced by a 12-week high-fat diet on Sar1b gene expression was also assessed by qRT-PCR.
Evaluation of Sar1b mRNA revealed the skeletal muscle as the tissue with the highest Sar1b expression, followed by the heart and liver, the organs composing the digestive tract, the brain and finally the lung and the adipose tissue. Sar1b protein expression levels follow a similar pattern among the organs, except for its lower expression in the heart. While the high-fat diet did not exert any significant alterations, Sar1b transgenic mice displayed higher gene expression in the liver, ileum, jejunum, proximal and distal colon compared to wild-type mice.
Our study supports the importance of Sar1b in organs involved in lipid transport and/or calcium trafficking such as the liver, intestine, skeletal muscle and heart.
背景/目的:Sar1b GTP酶(Sar1b)是COPII囊泡的一个必需组成部分,COPII囊泡从内质网出芽以将蛋白质运输到高尔基体。其基因突变会导致一种严重疾病,即乳糜微粒滞留病。尽管对Sar1b的了解越来越多,但对其组织分布和调控却知之甚少,而这正是本研究的目的。我们旨在通过与野生型小鼠对比,利用转基因小鼠确定Sar1b的组织分布以及高脂饮食和基因强迫对其的调节作用。方法:通过定量逆转录聚合酶链反应(qRT-PCR)和蛋白质免疫印迹法研究野生型和Sar1b转基因小鼠不同器官中Sar1b的表达模式。还通过qRT-PCR评估转基因和12周高脂饮食诱导的胰岛素抵抗对Sar1b基因表达的影响。
对Sar1b信使核糖核酸(mRNA)的评估显示,骨骼肌是Sar1b表达最高的组织,其次是心脏和肝脏、构成消化道的器官、大脑,最后是肺和脂肪组织。Sar1b蛋白表达水平在各器官中呈现类似模式,但在心脏中的表达较低。虽然高脂饮食未产生任何显著变化,但与野生型小鼠相比,Sar1b转基因小鼠在肝脏、回肠、空肠、近端和远端结肠中的基因表达更高。
我们的研究支持了Sar1b在参与脂质运输和/或钙运输的器官(如肝脏、肠道、骨骼肌和心脏)中的重要性。