Dick A P, Harik S I
J Neurochem. 1986 May;46(5):1406-11. doi: 10.1111/j.1471-4159.1986.tb01755.x.
We used [3H]cytochalasin B as a specific ligand to study the glucose transporter of the following tissue preparations: (a) microvessels derived from the cerebral cortex and cerebellum of the rat and pig, (b) particulate fractions of the cerebral cortex and cerebellum of the rat and pig, (c) lateral, third, and fourth ventricular choroid plexus of the pig, and (d) synaptosomes from the pig cerebral cortex. Specific, D-glucose-displaceable binding of [3H]cytochalasin B was present in all the preparations studied. This binding was saturable and displayed the kinetics of a single class of binding sites, similar to the glucose transporter found in other mammalian tissues. The density of the glucose transporter was much higher in cerebral and cerebellar microvessels and choroid plexus than either in crude particulate fractions of the cerebrum and cerebellum or in cerebral synaptosomes. These findings agree with the physiologic function of brain microvessels that transport glucose, not only for their own use, but also for the much greater mass of the entire brain. In the pig, the density of the glucose transporter in cerebral microvessels was significantly higher than in cerebellar microvessels. Irreversible photoaffinity labeling of the glucose transporter of synaptosomal membranes with [3H]cytochalasin B followed by solubilization and polyacrylamide gel electrophoresis demonstrated a single region of radioactivity that corresponded to a molecular mass of 60,000-64,000 daltons.
我们使用[3H]细胞松弛素B作为特异性配体,来研究以下组织制剂中的葡萄糖转运体:(a) 源自大鼠和猪大脑皮层及小脑的微血管;(b) 大鼠和猪大脑皮层及小脑的微粒体部分;(c) 猪的侧脑室、第三脑室和第四脑室脉络丛;(d) 猪大脑皮层的突触体。在所研究的所有制剂中均存在[3H]细胞松弛素B的特异性、可被D-葡萄糖置换的结合。这种结合具有饱和性,并表现出单一类结合位点的动力学特征,类似于在其他哺乳动物组织中发现的葡萄糖转运体。大脑和小脑微血管以及脉络丛中葡萄糖转运体的密度,比大脑和小脑的粗微粒体部分或大脑突触体中的密度高得多。这些发现与脑微血管的生理功能相符,脑微血管不仅运输葡萄糖供自身使用,还为整个大脑的大量组织运输葡萄糖。在猪中,大脑微血管中葡萄糖转运体的密度显著高于小脑微血管。用[3H]细胞松弛素B对突触体膜的葡萄糖转运体进行不可逆光亲和标记,随后进行溶解和聚丙烯酰胺凝胶电泳,结果显示出一个单一的放射性区域,其对应的分子量为60,000 - 64,000道尔顿。