Homma S, Gapstur S M, Coffey A, Valtin H, Dousa T P
Department of Medicine, Mayo Clinic and Foundation, Rochester, Minnesota 55905.
Am J Physiol. 1991 Aug;261(2 Pt 2):F345-53. doi: 10.1152/ajprenal.1991.261.2.F345.
To test the hypothesis that rapid adenosine 3',5'-cyclic monophosphate (cAMP) catabolism via cyclic 3',5'-nucleotide phosphodiesterase (PDE) is a cause of the unresponsiveness to vasopressin (VP) in mice with hereditary nephrogenic diabetes insipidus (NDI), we investigated properties of PDEs and other aspects of the VP-dependent cAMP-signaling system in segments of collecting ducts [inner medullary (IMCD), cortical (CCD), and outer medullary (OMCD) ducts] microdissected from control mice and mice with NDI. The activity of cAMP-PDE, but not of cGMP-PDE, was markedly higher in IMCD (+109%), and to a lesser degree in OMCD (+41%) and CCD (+27%), of NDI mice than in normal controls. The cAMP-PDE in IMCD of NDI mice was more sensitive to inhibition by the PDE isozyme-specific inhibitors rolipram and cilostamide, but not by 3-isobutyl-1-methylxanthine, than was the cAMP-PDE in controls. Levels of cAMP in intact IMCD and CCD from NDI mice completely failed to increase in response to 10(-6) M VP. Incubation with rolipram alone, but not with cilostamide alone, restored VP-dependent cAMP accumulation in IMCD of NDI mice to the levels found in control mice; addition of cilostamide further enhanced the effect of rolipram. Analogous (but quantitatively lesser) anomalies of the VP-dependent cAMP system, including the effects of PDE inhibitors, were observed also in CCD of NDI mice. However, the activity of VP-stimulated adenylate cyclase assayed in permeabilized IMCD did not differ in NDI and control mice. These results indicate that anomalously high activities of low-Km cAMP-PDE isozymes account for the failure of collecting ducts of NDI mice to increase cAMP levels in response in VP.(ABSTRACT TRUNCATED AT 250 WORDS)
即通过环3',5'-核苷酸磷酸二酯酶(PDE)介导的快速3',5'-环磷酸腺苷(cAMP)分解代谢是遗传性肾性尿崩症(NDI)小鼠对血管加压素(VP)无反应的原因,我们研究了从对照小鼠和NDI小鼠显微解剖得到的集合管节段[内髓集合管(IMCD)、皮质集合管(CCD)和外髓集合管(OMCD)]中PDE的特性以及VP依赖的cAMP信号系统的其他方面。与正常对照相比,NDI小鼠的IMCD中cAMP-PDE的活性显著更高(升高了109%),OMCD(升高了41%)和CCD(升高了27%)中的活性升高程度较小。NDI小鼠IMCD中的cAMP-PDE比对照小鼠中的cAMP-PDE对PDE同工酶特异性抑制剂咯利普兰和西洛酰胺的抑制作用更敏感,但对3-异丁基-1-甲基黄嘌呤不敏感。NDI小鼠完整的IMCD和CCD中的cAMP水平在10(-6) M VP刺激下完全没有升高。单独用咯利普兰孵育可使NDI小鼠IMCD中VP依赖的cAMP积累恢复到对照小鼠中的水平,而单独用西洛酰胺则不能;添加西洛酰胺可进一步增强咯利普兰的作用。在NDI小鼠的CCD中也观察到了VP依赖的cAMP系统类似(但数量上较少)的异常,包括PDE抑制剂的作用。然而,在透化的IMCD中检测的VP刺激的腺苷酸环化酶活性在NDI小鼠和对照小鼠中没有差异。这些结果表明,低Km cAMP-PDE同工酶的异常高活性是NDI小鼠集合管对VP反应时cAMP水平无法升高的原因。(摘要截短于250字)