Hara M, Matsuda Y, Hirai K, Okumura N, Nakagawa H
Institute for Protein Research, Osaka University, Japan.
J Neurochem. 1989 Mar;52(3):902-8. doi: 10.1111/j.1471-4159.1989.tb02540.x.
Glucose transport systems in cultured neuronal cells and astrocytes of rats were characterized by measuring the uptake of 2-deoxy-D-[3H]glucose ([3H]2-DG) into the cells. Various sugars inhibited 2-DG uptake by neuronal cells and astrocytes similarly, a finding indicating that the substrate specificities of the transporters in the two types of cells were almost the same. However, the Km values for 2-DG of neuronal cells and astrocytes were 1.7 and 0.36 mM, respectively. The uptake of 2-DG was strongly inhibited by cytochalasin B. Nucleosides, such as adenosine, inosine, and uridine, inhibited 2-DG uptake competitively in both neuronal cells and astrocytes. The uptake by both types of cells were also inhibited by forskolin, but not by cyclic AMP, an observation suggesting that forskolin bound directly to the transporters to cause inhibition. Its inhibition was competitive in astrocytes and noncompetitive in neuronal cells. Astrocytes contained a glucose transporter with a subunit molecular weight of 45K, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis after photoaffinity labeling using [3H]cytochalasin B as a probe.
通过测量2-脱氧-D-[3H]葡萄糖([3H]2-DG)进入细胞的摄取量,对大鼠培养的神经元细胞和星形胶质细胞中的葡萄糖转运系统进行了表征。各种糖类对神经元细胞和星形胶质细胞摄取2-DG的抑制作用相似,这一发现表明两种细胞中转运体的底物特异性几乎相同。然而,神经元细胞和星形胶质细胞对2-DG的Km值分别为1.7和0.36 mM。2-DG的摄取受到细胞松弛素B的强烈抑制。核苷,如腺苷、肌苷和尿苷,在神经元细胞和星形胶质细胞中均竞争性抑制2-DG的摄取。两种细胞的摄取也受到福斯高林的抑制,但不受环磷酸腺苷的抑制,这一观察结果表明福斯高林直接与转运体结合导致抑制。其抑制作用在星形胶质细胞中是竞争性的,在神经元细胞中是非竞争性的。使用[3H]细胞松弛素B作为探针进行光亲和标记后,通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳估计,星形胶质细胞含有一种亚基分子量为45K的葡萄糖转运体。