Grignon S, Seagar M J, Couraud F
Neurobiologie des Canaux Ioniques, INSERM U374, Faculté de Medecine, Marseille, France.
Neurosci Lett. 1993 May 28;155(1):87-91. doi: 10.1016/0304-3940(93)90679-f.
We have studied the biochemical and functional aspects of omega-conotoxin GVIA (omega-CgTx)-sensitive calcium channels in cerebellar granule cells in vitro. 125I-omega-Conotoxin GVIA (125I-omega-CgTx) binding sites were detected in intact cultured cerebellar granule cells and binding parameters were measured (Bmax: 134 fmol/mg protein; kinetic association constant kappa: 3.10(6) M-1.s-1). [3H]Glutamate release was assessed under different release paradigms (namely release triggered by calcium, voltage, and sodium channel agonists) and different times (15 s and 2 min). However, in all cases, [3H]glutamate release was found to be completely insensitive to omega-CgTx. Conversely, voltage-dependent release was inhibited in a dose-dependent fashion by cadmium chloride, with total inhibition at 10(-4) M. These results indicate that N-type calcium channels are not involved in glutamate secretion from granule neurons.
我们在体外研究了小脑颗粒细胞中对ω-芋螺毒素GVIA(ω-CgTx)敏感的钙通道的生化和功能方面。在完整的培养小脑颗粒细胞中检测到125I-ω-芋螺毒素GVIA(125I-ω-CgTx)结合位点,并测量了结合参数(Bmax:134 fmol/mg蛋白质;动力学缔合常数κ:3.10(6) M-1·s-1)。在不同的释放模式(即由钙、电压和钠通道激动剂触发的释放)和不同时间(15秒和2分钟)下评估了[3H]谷氨酸的释放。然而,在所有情况下,发现[3H]谷氨酸的释放对ω-CgTx完全不敏感。相反,电压依赖性释放被氯化镉以剂量依赖性方式抑制,在10(-4) M时完全抑制。这些结果表明N型钙通道不参与颗粒神经元的谷氨酸分泌。