Yuasa S, Takamitsu Y, Miki S, Orita Y, Abe H, Watanabe T
J Pharmacol Exp Ther. 1980 Oct;215(1):254-8.
Involvement of microtubules in the cellular action of vasopressin was studied by physiological and biochemical procedures. The inhibitory effect of 10(-4) M colchicine on the hydro-osmotic response of the toad bladder to 2 mM cyclic AMP was completely blocked by an increase in the concentration of calcium in the serosal bathing medium from 1 to 8 mM. Most of the colchicine-binding activity of extracts of the bladder epithelial cells was recovered by DEAE-Sephadex chromatography in a procedure similar to that used to isolate tubulin of bovine brain. Sodium dodecyl sulfate urea and urea gel electrophoresis of the DEAE-Sephadex fraction from the bladder epithelial cells yielded bands that corresponded to those of bovine brain tubulin. When 8 mM CaCl2 was present in an incubation mixture, the binding of colchicine to tubulin from the bladder epithelial cells and bovine brain was significantly inhibited. However, 1 mM CaCl2 did not affect the binding of colchicine to tubulin. Although the present study supports the concept that microtubules and calcium play an important role in the cellular action of vasopressin on the toad urinary bladder, the mechanism by which calcium blocks the inhibitory effect of colchicine on the hydro-osmotic response to cyclic AMP could not be elucidated.
通过生理学和生物化学方法研究了微管在抗利尿激素细胞作用中的参与情况。10(-4)M秋水仙碱对蟾蜍膀胱对2mM环磷酸腺苷的水渗透反应的抑制作用，被浆膜浴液中钙浓度从1mM增加到8mM完全阻断。膀胱上皮细胞提取物的大部分秋水仙碱结合活性，通过DEAE-葡聚糖凝胶色谱法得以恢复，其过程类似于用于分离牛脑微管蛋白的方法。膀胱上皮细胞DEAE-葡聚糖级分的十二烷基硫酸钠尿素和尿素凝胶电泳产生的条带，与牛脑微管蛋白的条带相对应。当孵育混合物中存在8mM氯化钙时，秋水仙碱与膀胱上皮细胞和牛脑微管蛋白的结合受到显著抑制。然而，1mM氯化钙并不影响秋水仙碱与微管蛋白的结合。尽管本研究支持微管和钙在抗利尿激素对蟾蜍膀胱的细胞作用中起重要作用这一概念，但钙阻断秋水仙碱对环磷酸腺苷水渗透反应抑制作用的机制仍无法阐明。
