脑内Glycine受体的结合特性与甾体激素相关性研究

Effect of Steroid on Binding Properties of Glycine Receptors on Synaptic Plasma Membrane from Rats

目的研究甾体激素对脑内Glycine受体结合的影响。方法制备大鼠脑突触质膜,排除胞液污染,用放射配体结合的方法,以[3H]甘氨酸为标记配体。结果测出甘氨酸受体结合的饱和曲线,Scatchard作图为一条直线,结合容量Bmax为每毫克蛋白(26±4)pmol,平衡解离常数Kd为(173±15)nmol.L-1,士的宁取代实验显示:脑干部甘氨酸受体为士的宁敏感型,其余脑部为士的宁不敏感型。甾体激素取代实验显示:孕酮对甘氨酸受体的结合有快速的抑制作用。结论孕激素对甘氨酸受体的结合有影响。

Objective To develop plasma membrane from rats. Methods effect of steroid on binding properties of glycine The synaptic membrane （SPM） was prepared receptors on synaptic from rat brain. There was no cytosolic contamination. The marked enzyme（LDH） was used to monitor the SPM purity. Results Using radioligand binding assay with [^3H] glycine as a ligand, a saturated curve of glycine receptor binding was obtained. The binding content（Bmax） was 26 ± 4 pmol/mg protein and the equilibrium dissociation constant （Kd） was 173 ±15 nmol· L^-1. the IC50 of glycine was 10^-6mol· L^-1 which was calculated from the displacement curve. The glycine receptor was strychnine - sensitive in the brainstem, but not in other brain regions. Condusion Inhibiting experiment with steroid hormones showed that progesterone have some effects on glycine receptor binding.