van Baal J, Raber G, de Slegte J, Pieters R, Bindels R J, Willems P H
Department of Cell Physiology, University of Nijmegen, PO Box 9101, NL-6500 HB Nijmegen, The Netherlands.
Pflugers Arch. 1996 Nov-Dec;433(1-2):109-15. doi: 10.1007/s004240050255.
The effect of arginine vasopressin (AVP) on transepithelial Ca2+ transport in primary cultures of rabbit cortical collecting system cells was examined. Addition of AVP to the basolateral side of the monolayer dose-dependently (EC50 = 0.7 nM) increased active Ca2+ reabsorption from a basal value of 85 +/- 2 nmol.h-1.cm-2 to a maximum value of 124 +/- 3 nmol.h-1.cm-2. This was paralleled by a dose-dependent (EC50 = 1.1 nM) increase in cellular adenosine 3', 5'-cyclic monophosphate (cAMP) content. Both effects of AVP were mimicked by the V2 agonist deamino-Cys,D-Arg8-vasopressin (dDAVP) and forskolin. Addition of either AVP or dDAVP to the basolateral side evoked a sustained increase in cytosolic free Ca2+ concentration, which resulted from both Ca2+ entry and release from internal stores. Only the effect on Ca2+ entry was mimicked by forskolin, demonstrating that cAMP acts by activating a Ca2+ influx pathway. The present findings demonstrate that AVP stimulates transcellular Ca2+ transport in the cortical collecting system through activation of basolateral V2 receptors coupled to adenylyl cyclase to increase the cellular cAMP content.
研究了精氨酸加压素(AVP)对兔皮质集合系统细胞原代培养物中跨上皮钙(Ca2+)转运的影响。将AVP添加到单层细胞的基底外侧,可剂量依赖性地(EC50 = 0.7 nM)增加活性Ca2+重吸收,从基础值85±2 nmol·h-1·cm-2增加到最大值124±3 nmol·h-1·cm-2。这与细胞内3',5'-环磷酸腺苷(cAMP)含量的剂量依赖性增加(EC50 = 1.1 nM)平行。AVP的这两种作用均被V2激动剂去氨基半胱氨酸、D-精氨酸8-加压素(dDAVP)和毛喉素模拟。将AVP或dDAVP添加到基底外侧均可引起胞质游离Ca2+浓度的持续升高,这是由Ca2+内流和从内部储存库释放所致。只有对Ca2+内流的作用被毛喉素模拟,表明cAMP通过激活Ca2+内流途径发挥作用。本研究结果表明,AVP通过激活与腺苷酸环化酶偶联的基底外侧V2受体来增加细胞内cAMP含量,从而刺激皮质集合系统中的跨细胞Ca2+转运。