Yamada Toshiki, Konno Norifumi, Matsuda Kouhei, Uchiyama Minoru
Department of Life and Environmental Science, Graduate School of Science and Engineering, University of Toyama, 3190 Gofuku, Toyama 930-8555, Japan.
J Comp Physiol B. 2007 Jul;177(5):503-8. doi: 10.1007/s00360-007-0148-5. Epub 2007 Feb 16.
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.
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。