Yu S, Ding W G
Institute of Molecular Neurobiology, Shiga University of Medical Science, Otsu 520-2192, Japan.
Brain Res. 1998 Jun 22;797(1):65-72. doi: 10.1016/s0006-8993(98)00372-2.
We and others have previously reported that glucose transporter 1 (GLUT1)-like 45 kDa protein is localized to parenchymal cells in the brain. However, the precise cellular localization has remained unclear. In the present study, we examined the cellular localization of GLUT1 in the rat brain by double immunostaining methods and immunoelectron microscopic analysis using a rabbit antiserum specific to GLUT1. Western blot analysis of the rat brain revealed that the antiserum detected a strong band with a molecular weight of 45 kDa and a weak band of about 55 kDa, which corresponded respectively to the known molecular weights of the GLUT1 proteins in the brain parenchymal cells and the brain microvessels. Immunohistochemical staining revealed a large number of GLUT1-immunoreactive glial cells and microvessels in almost every region of the brain. Double immunofluorescence analysis demonstrated that the GLUT1-like 45 kDa protein occurred in many galactocerebroside-positive oligodendrocytes and in some glial fibrillary acidic protein (GFAP)-positive astrocytes. No GLUT1-immunoreactivity was observed in OX42-positive microglia. Immunoelectron microscopic examination confirmed that the GLUT1-immunoreactivity was mainly localized in the cytoplasm of the oligodendrocytes and astrocytes. The results indicate that the 45 kDa form of GLUT1 protein exists in the glial cells including astrocytes and oligodendrocytes.
我们及其他研究人员之前曾报道,葡萄糖转运蛋白1(GLUT1)样45 kDa蛋白定位于脑实质细胞。然而,其精确的细胞定位仍不清楚。在本研究中,我们通过双重免疫染色方法及使用针对GLUT1的兔抗血清进行免疫电子显微镜分析,研究了GLUT1在大鼠脑中的细胞定位。对大鼠脑进行的蛋白质印迹分析显示,该抗血清检测到一条分子量为45 kDa的强条带和一条约55 kDa的弱条带,它们分别对应于脑实质细胞和脑微血管中已知分子量的GLUT1蛋白。免疫组织化学染色显示,在脑的几乎每个区域都有大量GLUT1免疫反应性胶质细胞和微血管。双重免疫荧光分析表明,45 kDa的GLUT1样蛋白存在于许多半乳糖脑苷脂阳性少突胶质细胞及一些胶质纤维酸性蛋白(GFAP)阳性星形胶质细胞中。在OX42阳性小胶质细胞中未观察到GLUT1免疫反应性。免疫电子显微镜检查证实,GLUT1免疫反应性主要定位于少突胶质细胞和星形胶质细胞的细胞质中。结果表明,45 kDa形式的GLUT1蛋白存在于包括星形胶质细胞和少突胶质细胞在内的胶质细胞中。