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日本树蛙(Hyla japonica)膀胱细胞中的蛙心钠素和环鸟苷酸激活氨氯地平敏感的钠离子通道

Frog atrial natriuretic peptide and cGMP activate amiloride-sensitive Na(+) channels in urinary bladder cells of Japanese tree frog, Hyla japonica.

作者信息

Yamada Toshiki, Konno Norifumi, Matsuda Kouhei, Uchiyama Minoru

机构信息

Department of Life and Environmental Science, Graduate School of Science and Engineering, University of Toyama, 3190 Gofuku, Toyama 930-8555, Japan.

出版信息

J Comp Physiol B. 2007 Jul;177(5):503-8. doi: 10.1007/s00360-007-0148-5. Epub 2007 Feb 16.

Abstract

In our previous study, it was suggested that ANP and cGMP may increase Na(+) absorption in the urinary bladder of the Japanese tree frog, Hyla japonica. Thus, Na(+) transport activated by ANP was investigated electrophysiologically by using a cell-attached patch-clamp technique in freshly isolated cells from the urinary bladder. A predominant channel expressed was a low conductance Na(+) channel in the epithelial cells. The channel exhibited conductance for inward currents of 4.9 +/- 0.2 pS, long open and closed times (c.a. 190 ms), and positive reversal potential. The channel activity was decreased under the pipette solution including 10(-6) M amiloride. These characteristics were similar to those of amiloride-sensitive Na(+) channels (ENaC). Addition of 10(-9) M ANP activated and significantly increased the ENaC activity from 0.58 +/- 0.09 to 1.47 +/- 0.34. On the other hand, mean amplitudes and conductance of single channel did not change significantly after the addition of ANP. Addition of 10(-5) M 8-Br-cGMP also activated the ENaC and significantly increased the channel activity from 0.56 +/- 0.10 to 2.00 +/- 0.33. The addition of ANP failed to activate the ENaC in the presence of 10(-6) M amiloride. These results suggested that ANP and cGMP activate Na(+) transport via ENaC in the epithelial cells of frog urinary bladder.

摘要

在我们之前的研究中,有人提出心房钠尿肽(ANP)和环磷酸鸟苷(cGMP)可能会增加日本树蛙(Hyla japonica)膀胱中的钠(Na⁺)吸收。因此,我们采用细胞贴附式膜片钳技术,在新鲜分离的膀胱细胞中,通过电生理学方法研究了ANP激活的Na⁺转运。上皮细胞中表达的主要通道是一种低电导Na⁺通道。该通道对内向电流的电导为4.9±0.2 pS,开放和关闭时间较长(约190毫秒),且反转电位为正。在含有10⁻⁶ M氨氯吡脒的移液管溶液中,该通道活性降低。这些特性与氨氯吡脒敏感的Na⁺通道(ENaC)相似。添加10⁻⁹ M ANP可激活ENaC,并使其活性从0.58±0.09显著增加至1.47±0.34。另一方面,添加ANP后,单通道的平均幅度和电导没有显著变化。添加10⁻⁵ M 8-溴-cGMP也可激活ENaC,并使其活性从0.56±0.10显著增加至2.00±0.33。在存在10⁻⁶ M氨氯吡脒的情况下,添加ANP无法激活ENaC。这些结果表明,ANP和cGMP通过蛙膀胱上皮细胞中的ENaC激活Na⁺转运。

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