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葡萄糖转运蛋白8(GLUT8)在小鼠肠道中的表达:可变剪接变体的鉴定。

Expression of GLUT8 in mouse intestine: identification of alternative spliced variants.

作者信息

Romero Amparo, Gomez Olga, Terrado Jose, Mesonero Jose E

机构信息

Department of Physiology, Pharmacology and Toxicology, Experimental Sciences and Health Faculty, University CEU - Cardenal Herrera, E-46113 Moncada, Valencia, Spain.

出版信息

J Cell Biochem. 2009 Apr 15;106(6):1068-78. doi: 10.1002/jcb.22090.

Abstract

GLUT8 is a facilitative glucose transporter composed of 10 exons coding for a 477 amino acids protein. It is mainly expressed in the testis, but it has also been studied in a number of tissues such as brain, adipose tissue, and liver. In this work, we have characterized the expression of GLUT8 in the small and large intestine under normal physiological conditions. Protein assay revealed low GLUT8 protein levels in the intestine compared to the testis, with higher levels in the colon than in the small intestine. Immunohistochemistry studies showed an intracellular localization of GLUT8 in enterocytes and colonocytes with a supranuclear distribution next to the apical membrane. GLUT8 immunoreactivity was also detected in the crypt cells. Interestingly, we have identified three additional transcriptional variants in mouse intestine (mGLUT-SP1, mGLUT8-SP2, and mGLUT8-SP3) produced by the deletion of one, two, and four exons, respectively, whereas only the entire mRNA was detected in the testis. Expression of these alternative variants did not have an effect on glucose consumption in 3T3-L1 cells. Although the specific function of GLUT8 in intestine remains unclear, the alternative splicing of GLUT8 could reflect a mechanism for the regulation of the gene expression in a tissue-specific manner by targeting GLUT8 mRNA for nonsense-mediated decay.

摘要

葡萄糖转运蛋白8(GLUT8)是一种易化型葡萄糖转运体,由10个外显子组成,编码一个含477个氨基酸的蛋白质。它主要在睾丸中表达,但也在许多组织中得到研究,如脑、脂肪组织和肝脏。在这项研究中,我们对正常生理条件下小肠和大肠中GLUT8的表达进行了特征分析。蛋白质检测显示,与睾丸相比,肠道中GLUT8蛋白水平较低,结肠中的水平高于小肠。免疫组织化学研究表明,GLUT8在肠上皮细胞和结肠细胞内定位,在顶端膜旁呈核上分布。在隐窝细胞中也检测到了GLUT8免疫反应性。有趣的是,我们在小鼠肠道中鉴定出另外三种转录变体(mGLUT8-SP1、mGLUT8-SP2和mGLUT8-SP3),它们分别是由缺失一个、两个和四个外显子产生的,而在睾丸中仅检测到完整的mRNA。这些可变变体的表达对3T3-L1细胞中的葡萄糖消耗没有影响。尽管GLUT8在肠道中的具体功能仍不清楚,但GLUT8的可变剪接可能反映了一种通过将GLUT8 mRNA靶向无义介导的衰变以组织特异性方式调节基因表达的机制。

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