Reagan L P, Gorovits N, Hoskin E K, Alves S E, Katz E B, Grillo C A, Piroli G G, McEwen B S, Charron M J
Harold and Margaret Milliken Hatch Laboratory of Neuroendocrinology, The Rockefeller University, 1230 York Avenue, New York, NY 10021, USA.
Proc Natl Acad Sci U S A. 2001 Feb 27;98(5):2820-5. doi: 10.1073/pnas.051629798. Epub 2001 Feb 13.
We describe the localization of the recently identified glucose transporter GLUTx1 and the regulation of GLUTx1 in the hippocampus of diabetic and control rats. GLUTx1 mRNA and protein exhibit a unique distribution when compared with other glucose transporter isoforms expressed in the rat hippocampus. In particular, GLUTx1 mRNA was detected in hippocampal pyramidal neurons and granule neurons of the dentate gyrus as well as in nonprincipal neurons. With immunohistochemistry, GLUTx1 protein expression is limited to neuronal cell bodies and the most proximal dendrites, unlike GLUT3 expression that is observed throughout the neuropil. Immunoblot analysis of hippocampal membrane fractions revealed that GLUTx1 protein expression is primarily localized to the intracellular compartment and exhibits limited association with the plasma membrane. In streptozotocin diabetic rats compared with vehicle-treated controls, quantitative autoradiography showed increased GLUTx1 mRNA levels in pyramidal neurons and granule neurons; up-regulation of GLUTx1 mRNA also was found in nonprincipal cells, as shown by single-cell emulsion autoradiography. In contrast, diabetic and control rats expressed similar levels of hippocampal GLUTx1 protein. These results indicate that GLUTx1 mRNA and protein have a unique expression pattern in rat hippocampus and suggest that streptozotocin diabetes increases steady-state mRNA levels in the absence of concomitant increases in GLUTx1 protein expression.
我们描述了最近鉴定出的葡萄糖转运蛋白GLUTx1在糖尿病大鼠和对照大鼠海马体中的定位以及GLUTx1的调节情况。与大鼠海马体中表达的其他葡萄糖转运蛋白亚型相比,GLUTx1 mRNA和蛋白呈现出独特的分布。特别是,在海马锥体细胞、齿状回颗粒细胞以及非主要神经元中检测到了GLUTx1 mRNA。通过免疫组织化学方法发现,与在整个神经纤维网中观察到的GLUT3表达不同,GLUTx1蛋白表达仅限于神经元细胞体和最靠近的树突。对海马膜组分的免疫印迹分析显示,GLUTx1蛋白表达主要定位于细胞内区室,与质膜的结合有限。与用载体处理的对照相比,在链脲佐菌素诱导的糖尿病大鼠中,定量放射自显影显示锥体细胞和颗粒细胞中GLUTx1 mRNA水平升高;单细胞乳胶放射自显影显示,非主要细胞中也发现了GLUTx1 mRNA的上调。相比之下,糖尿病大鼠和对照大鼠海马GLUTx1蛋白表达水平相似。这些结果表明,GLUTx1 mRNA和蛋白在大鼠海马体中具有独特的表达模式,并提示链脲佐菌素诱导的糖尿病在GLUTx1蛋白表达没有相应增加的情况下会增加稳态mRNA水平。