Integrated Bioscience Section, Graduate School of Science and Technology, Shizuoka University, Shizuoka 422-8529, Japan.
J Exp Biol. 2010 Jan 15;213(2):288-94. doi: 10.1242/jeb.036871.
The ventral pelvic skin of the tree frog Hyla japonica expresses two kinds of arginine vasotocin (AVT)-stimulated aquaporins (AQP-h2 and AQP-h3), which affect the capacity of the frog's skin to absorb water. As such, it can be used as a model system for analyzing the molecular mechanisms of water permeability. We investigated AQP dynamics and water permeability in the pelvic skin of H. japonica following challenge with AVT, hydrins (intermediate peptides of pro-AVT) and beta-adrenergic effectors. In the in vivo experiment, both AQP-h2 and AQP-h3 proteins were translocated to the apical plasma membrane in the principal cells of the first-reacting cell (FRC) layer in the pelvic skin following challenge with AVT, hydrin 1 and hydrin 2, thereby increasing the water permeability of the pelvic skin. The beta-adrenergic receptor agonist isoproterenol (IP) and its anatagonist propranolol (PP) in combination with AVT or hydrins were used as challenge in the in vitro experiment. IP increased water permeability whereas PP inhibited it, and both events were well correlated with the translocation of the AQPs to the apical membrane. In the PP+AVT-treated skins, labels for AQP-h2 and AQP-h3 were differentially visible among the principal cells; the apical plasma membrane of some cells was labeled while others were not, indicating that the response of PP or AVT is different from cell to cell. These results provide morphological evidence that the principal cells of the FRC layers may have two kinds of receptors: a V2 receptor and beta-adrenergic receptor.
树蛙 Hyla japonica 的腹侧骨盆皮肤表达两种精氨酸加压素(AVT)刺激的水通道蛋白(AQP-h2 和 AQP-h3),这会影响青蛙皮肤吸收水分的能力。因此,它可以作为分析水通透性分子机制的模型系统。我们研究了 AVT、hydrins(pro-AVT 的中间肽)和β-肾上腺素能效应物刺激后,H. japonica 骨盆皮肤中的 AQP 动力学和水通透性。在体内实验中,AVT、hydrin 1 和 hydrin 2 刺激后,AQP-h2 和 AQP-h3 蛋白均被转运到骨盆皮肤第一层反应细胞(FRC)主细胞的顶质膜,从而增加了骨盆皮肤的水通透性。β-肾上腺素能受体激动剂异丙肾上腺素(IP)及其拮抗剂普萘洛尔(PP)与 AVT 或 hydrins 联合用于体外实验。IP 增加水通透性,而 PP 抑制水通透性,这两种作用均与 AQP 向顶质膜的转运密切相关。在 PP+AVT 处理的皮肤中,AQP-h2 和 AQP-h3 的标记在主细胞之间可见差异;一些细胞的顶质膜被标记,而其他细胞则没有,表明 PP 或 AVT 的反应在细胞间存在差异。这些结果提供了形态学证据,表明 FRC 层的主细胞可能具有两种受体:V2 受体和β-肾上腺素能受体。