Emilio M G, Menano H P
Biochim Biophys Acta. 1975 Mar 25;382(3):344-52. doi: 10.1016/0005-2736(75)90276-x.
H+ extrusion by the isolated skins of two amphibia, Rana ridibunda and Bufo bufo, was studied in order to test for the presence of exchange mechanisms of the type Na+/H+ and Cl-/HCO3-, which have been described in several epithelial structures. The preparations were mounted in chambers of the Ussing type, so that the short-circuit current could be used as a function of Na+ transport and the pH-stat techinique was utilize to determine the rates of H+ extrusion under different experimental conditions. The conditions were either the withdrawal of the ions intervening the mentioned exchanges (Cl- or Na+), or the addition of drugs with well-known effects on Na+ up-take and transport (antidiuretic hormone and amiloride). In the frog skin, H+ excretion was detected in solutions containing either Cl- or SO4-2-, with identical rates. Again, Na+ substitution by Mg-2+ had no effect on H+ excretion rates, neither did the suppression of Na+ influx by amiloride or its stimulation by antidiuretic hormone. These experiments were repeated with similar results in gland-free preparations of the epidermis of frog skin separated from the corion by the action of collagenase. Experiments in toad skin that H+ excretion could not be detected whan Cl- was present in the outer medium, but became apparent if an impermant anion, SO4-2-, was used. This observation is compatible with the existence of an exchange mechanism of the type Cl-/HCO3-. Secondly, in these preparations H+ extrusion increased after stimulation with antidiuretic hormone and decreased when amiloride was used or when Na+ was substituted by Mg+, suggesting that a least a fraction of the total H+ efflux is linked to Na+ influx. In the isolated frog skin this mechanism does not seem to be operative.
为了测试在几种上皮结构中已描述过的Na⁺/H⁺和Cl⁻/HCO₃⁻类型的交换机制是否存在,对两种两栖动物即食用蛙和蟾蜍的离体皮肤的H⁺排出情况进行了研究。将标本安装在Ussing型小室中,以便短路电流可以用作Na⁺转运的函数,并利用pH计技术来确定在不同实验条件下H⁺的排出速率。这些条件要么是去除参与上述交换的离子(Cl⁻或Na⁺),要么是添加对Na⁺摄取和转运有已知作用的药物(抗利尿激素和氨氯吡脒)。在蛙皮中,在含有Cl⁻或SO₄²⁻的溶液中均检测到了H⁺排泄,速率相同。同样,用Mg²⁺替代Na⁺对H⁺排泄速率没有影响,氨氯吡脒抑制Na⁺内流或抗利尿激素刺激Na⁺内流也对H⁺排泄速率没有影响。在用胶原酶作用从真皮分离的蛙皮无腺体表皮制剂中重复了这些实验,结果相似。在蟾蜍皮肤的实验中,当外部介质中存在Cl⁻时无法检测到H⁺排泄,但如果使用非永久性阴离子SO₄²⁻,H⁺排泄就会变得明显。这一观察结果与Cl⁻/HCO₃⁻类型的交换机制的存在是相符的。其次,在这些制剂中,抗利尿激素刺激后H⁺排出增加,使用氨氯吡脒时或用Mg⁺替代Na⁺时H⁺排出减少,这表明总H⁺外流中至少有一部分与Na⁺内流有关。在离体蛙皮中,这种机制似乎不起作用。