MECHANISM OF DESENSITIZATION OF THE CLONED VASOPRESSIN V-1A RECEPTOR EXPRESSED IN XENOPUS-OOCYTES

被引:12
作者
NATHANSON, MH [1 ]
BURGSTAHLER, AD [1 ]
ORLOFF, JJ [1 ]
MANI, A [1 ]
MOYER, MS [1 ]
机构
[1] YALE UNIV,SCH MED,DEPT PEDIAT,NEW HAVEN,CT 06510
来源
AMERICAN JOURNAL OF PHYSIOLOGY | 1994年 / 267卷 / 01期
关键词
CONFOCAL MICROSCOPY; PROTEIN KINASE C; ADENOSINE; 3'; 5'-CYCLIC MONOPHOSPHATE; CYTOSOLIC CALCIUM;
D O I
10.1152/ajpcell.1994.267.1.C94
中图分类号
Q4 [生理学];
学科分类号
071003 ;
摘要
The vasopressin V-1a receptor exerts its effects by G protein-mediated increases in cytosolic Ca2+ (Ca-i(2a+)) and activation of protein kinase C. The V-1a, receptor also undergoes autologous desensitization. To clarify the mechanism of this desensitization, we expressed the cloned receptor in Xenopus oocytes, and vasopressin-induced Ca-i(2+) waves were examined as an index of V-1a, activation using confocal microscopy. Pretreatment of oocytes with a minimal concentration of vasopressin inhibited further generation of Ca-i(2+) waves upon maximal stimulation. Such pretreatment did not abolish Ca-i(2+) waves induced by subsequent microinjection of inositol trisphosphate, suggesting that this phenomenon represents receptor desensitization rather than depletion of inositol trisphosphate-sensitive Ca-i(2+) stores. Pretreatment with phorbol dibutyrate, ionomycin, or 8-bromoadenosine 3',5'-cyclic monophosphate had no effect on vasopressin-induced Ca-i(2+) waves. Oocytes recovered from desensitization within 1 h, but the microtubule inhibitor methyl-5-[2-thienylcarbonyl]-1H-benzimiidazol-2-yl)-carbamate (nocodazole) inhibited this recovery. Receptor binding sites were reduced by over 50% within 10 min of exposure to vasopressin, with no associated change in the K-d for the V-1a, receptor. These findings indicate that 1) expression of the cloned V-1a receptor in Xenopus oocytes, coupled with subcellular Ca-i(2+) imaging, provides a useful system to examine mechanisms of V-1a desensitization, 2) the V-1a receptor undergoes autologous desensitization in this experimental system, and 3) protein kinase C, Ca-i(2+), and adenosine 3',5'-cyclic monophosphate do not appear responsible for this desensitization, but 4) microtubule-dependent recycling of the receptor is preserved in this system and may be important for receptor desensitization.
引用
收藏
页码:C94 / C103
页数:10
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