Graves J S, Dunn B E, Brown S C
Am J Physiol. 1986 Nov;251(5 Pt 1):C787-94. doi: 10.1152/ajpcell.1986.251.5.C787.
By removing the shell membranes from the chorioallantoic membrane, the chorion is damaged, as visualized by electron microscopy, and rendered permeable, as evidenced by penetration of horseradish peroxidase and increased inhibition of the allantoic Na+-K+ pump by ouabain applied on the chorionic side. The short-circuit current (SCC) of this functionally isolated allantoic epithelium is augmented by nystatin, a channel-forming ionophore, when applied to the mucosal surface. Electrical parameters were determined for three age groups between 12 and 19 days of incubation. The SCC approximately doubled from the youngest (12-13 days) to the oldest (18-19 days) groups, whereas the transepithelial resistance (Re) of 700-900 omega X cm2 remained the same. Amiloride, an inhibitor of apical Na+ uptake, inhibited 98-100% of the SCC at 10(-4) M in both 15-16 and 18-19 day epithelia. In the 12- to 13-day preparation 20-25% of the SCC was insensitive to 10(-3) M amiloride. The Ki's for amiloride were similar in all preparations, at about 5 X 10(-7) M. Determination of the Hill coefficients for inhibition revealed a lower value (0.75 +/- 0.03) for the 12-13 day preparation compared with the two older preparations with coefficients not significantly different from unity. Replacing Na+ in the bathing solutions abolished the SCC of 18-19 day epithelia, whereas about 15% of the SCC remained at 12-13 days. Thus, during development, the SCC of the allantoic epithelium increases in magnitude and becomes increasingly (to 100%) amiloride-sensitive and Na+-dependent.
通过去除尿囊绒毛膜的壳膜,绒毛膜会受到损伤,这在电子显微镜下可见,并且变得具有通透性,这可由辣根过氧化物酶的渗透以及应用于绒毛膜侧的哇巴因对尿囊钠钾泵抑制作用的增强来证明。当制霉菌素(一种形成通道的离子载体)应用于黏膜表面时,这种功能分离的尿囊上皮的短路电流(SCC)会增加。测定了孵化12至19天之间三个年龄组的电参数。SCC从最年轻的组(12 - 13天)到最年长的组(18 - 19天)大约增加了一倍,而跨上皮电阻(Re)在700 - 900Ω×cm²保持不变。氨氯吡脒(一种顶端钠摄取抑制剂)在10⁻⁴M时,对15 - 16天和18 - 19天的上皮细胞抑制了98 - 100%的SCC。在12至13天的制剂中,20 - 25%的SCC对10⁻³M氨氯吡脒不敏感。在所有制剂中氨氯吡脒的抑制常数(Ki)相似,约为5×10⁻⁷M。抑制作用的希尔系数测定显示,与另外两个系数与1无显著差异的较年长制剂相比,12 - 13天制剂的值较低(0.75±0.03)。在浸泡溶液中用其他离子替代钠会消除18 - 19天上皮细胞的SCC,而在12 - 13天时仍有约15%的SCC。因此,在发育过程中,尿囊上皮的SCC在幅度上增加,并且对氨氯吡脒的敏感性和对钠的依赖性越来越高(达到100%)。