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大鼠周围神经的葡萄糖转运体。雪旺细胞和神经周细胞在体内和体外对GLUT1基因的差异表达。

Glucose transporters of rat peripheral nerve. Differential expression of GLUT1 gene by Schwann cells and perineural cells in vivo and in vitro.

作者信息

Muona P, Sollberg S, Peltonen J, Uitto J

机构信息

Department of Medical Biochemistry, University of Turku, Finland.

出版信息

Diabetes. 1992 Dec;41(12):1587-96. doi: 10.2337/diab.41.12.1587.

Abstract

Expression of GLUTs in rat peripheral nerve was first studied at the mRNA level with Northern transfer analysis with cDNAs specific for GLUT1, GLUT2, GLUT3, and GLUT4. GLUT1 mRNA was the only GLUT mRNA detectable in rat sciatic nerve. In situ hybridization localized this mRNA to the perineurium and to some endo- and epineurial capillaries. Indirect immunofluorescence stainings demonstrated that GLUT1 protein epitopes were concentrated primarily in the perineurium and endoneurial capillaries. Also, some Schwann cells, a few epineurial capillaries, and medium-sized blood vessels showed a faintly positive immunoreaction. All cell types present in primary cultures initiated from rat sciatic nerve (perineurial cells, Schwann cells, and fibroblasts) expressed GLUT1 protein in vitro. Thus, Schwann cells, which expressed GLUT1 only occasionally at a low level in vivo, have the potential to express GLUT1 at a markedly higher level under cell culture conditions. Incubation of the cultures in 25 mM D-glucose for 7 days caused a 39% reduction in the amount of immunodetectable GLUT1 protein, and a marked (34%) decrease of GLUT1 mRNA compared with cultures incubated in 5.5 mM D-glucose. Interestingly, the reduction of [3H]-2-DG uptake in the same cultures exceeded 70%, suggesting that the reduced amount of GLUT1 protein alone did not explain the marked reduction in glucose uptake in these cultures. Immunostaining of the cell cultures suggested that perineurial cells were the main target for the glucose-induced decrease of GLUT1 protein.

摘要

首先运用针对GLUT1、GLUT2、GLUT3和GLUT4的cDNA进行Northern印迹分析,在mRNA水平上研究大鼠外周神经中葡萄糖转运蛋白(GLUTs)的表达情况。GLUT1 mRNA是在大鼠坐骨神经中唯一可检测到的GLUT mRNA。原位杂交将该mRNA定位到神经束膜以及一些神经内膜和神经外膜毛细血管。间接免疫荧光染色表明,GLUT1蛋白表位主要集中在神经束膜和神经内膜毛细血管中。此外,一些施万细胞、少数神经外膜毛细血管和中等大小的血管呈现出弱阳性免疫反应。从大鼠坐骨神经原代培养的所有细胞类型(神经束膜细胞、施万细胞和成纤维细胞)在体外均表达GLUT1蛋白。因此,在体内仅偶尔以低水平表达GLUT1的施万细胞,在细胞培养条件下有潜力以明显更高的水平表达GLUT1。将培养物在25 mM D-葡萄糖中孵育7天,与在5.5 mM D-葡萄糖中孵育的培养物相比,可免疫检测到的GLUT1蛋白量减少了39%,GLUT1 mRNA显著减少(34%)。有趣的是,相同培养物中[3H]-2-DG摄取的减少超过70%,这表明仅GLUT1蛋白量的减少并不能解释这些培养物中葡萄糖摄取的显著减少。细胞培养物的免疫染色表明,神经束膜细胞是葡萄糖诱导的GLUT1蛋白减少的主要靶点。

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