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原代培养神经元和神经胶质细胞中GLUT1和GLUT3葡萄糖转运蛋白的免疫定位

Immunolocalization of GLUT1 and GLUT3 glucose transporters in primary cultured neurons and glia.

作者信息

Maher F

机构信息

National Institute of Diabetes, Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, USA.

出版信息

J Neurosci Res. 1995 Nov 1;42(4):459-69. doi: 10.1002/jnr.490420404.

DOI:10.1002/jnr.490420404
PMID:8568932
Abstract

Immunofluorescence analysis was used to study the cellular localization of glucose transporters 1 and 3 (GLUT1 and GLUT3) in primary rat neuronal and glial cultures. In primary cultured cerebellar granule neurons and cortical neurons, GLUT3 was detected in a pattern consistent with a generalized cell surface distribution. GLUT3 distribution corresponded most closely with the neural cell adhesion molecule (NCAM), and showed overlapping but distinct distributions compared to synaptophysin, microtubule-associated protein 2 (MAP2), neurofilament protein, and growth-associated protein (GAP43). Culture of neurons in the presence of glia did not alter the cellular localization of GLUT3. GLUT1 was detectable in primary cerebellar granule neurons both at the cell surface and in the cytoplasm, and appeared decreased in neurons cocultured with glia. GLUT1, but not GLUT3, was detected in glial fibrillary acidic protein (GFAP)-positive astrocytes present in mixed neuronal-glial cultures derived from cerebellum and cerebral cortex, as well as in cortical astrocyte cultures. GLUT1, but not GLUT3, was also detected in microglia and oligodendrocytes present in these cultures. This study indicates a generalized cell surface expression of the glucose transporters expressed in neurons and glia, rather than selective targeting to different cellular domains or subcellular locations.

摘要

免疫荧光分析用于研究原代大鼠神经元和神经胶质细胞培养物中葡萄糖转运蛋白1和3(GLUT1和GLUT3)的细胞定位。在原代培养的小脑颗粒神经元和皮质神经元中,检测到GLUT3的分布模式与普遍的细胞表面分布一致。GLUT3的分布与神经细胞粘附分子(NCAM)最为接近,与突触素、微管相关蛋白2(MAP2)、神经丝蛋白和生长相关蛋白(GAP43)相比,显示出重叠但不同的分布。在有神经胶质细胞存在的情况下培养神经元,不会改变GLUT3的细胞定位。在原代小脑颗粒神经元的细胞表面和细胞质中均可检测到GLUT1,并且在与神经胶质细胞共培养的神经元中,GLUT1的含量似乎有所下降。在源自小脑和大脑皮质的混合神经元-神经胶质细胞培养物中存在的胶质纤维酸性蛋白(GFAP)阳性星形胶质细胞中,以及在皮质星形胶质细胞培养物中,均可检测到GLUT1,而未检测到GLUT3。在这些培养物中的小胶质细胞和少突胶质细胞中也检测到了GLUT1,而未检测到GLUT3。这项研究表明,神经元和神经胶质细胞中表达的葡萄糖转运蛋白在细胞表面普遍表达,而不是选择性地靶向不同的细胞结构域或亚细胞位置。

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