Grady T P, Cheung L Y
Am J Physiol. 1983 Jan;244(1):G71-5. doi: 10.1152/ajpgi.1983.244.1.G71.
Intracellular microelectrode techniques were applied to Necturus antral mucosa. Stable intracellular impalements were obtained with 15-50 M omega microelectrodes filled with 3M KCl. It was possible to change rapidly the mucosal bathing solution while maintaining the microelectrode in the cell. With these techniques, we were able to measure the electrical potentials and resistances of the cell membranes and the shunt pathway. The transepithelial potential was -4.9 +/- 1.3 mV, serosal solution reference. Apical cell membrane potential was -43.9 +/- 0.6 mV, cell negative to the mucosal solution. Basolateral cell membrane potential was -48.8 +/- 1.3 mV, cell negative to serosal solution. Transepithelial resistance was 427 +/- 66 omega . cm2. The ratio of apical to basolateral membrane resistances was 3.4 +/- 0.3. The electrical resistances of the transcellular and paracellular pathway were determined by the measurement of the total transepithelial resistance and the ratio of apical to basolateral cell membrane resistances before and after blocking apical membrane sodium permeability with amiloride. The resistances of the apical cell membrane, basolateral cell membrane, and the shunt were 2,203 +/- 585, 1,296 +/- 384, and 604 +/- 81 omega . cm2, respectively (mean +/- SE). Calculations from these measurements indicate that the shunt contribution to transepithelial conductance was approximately 85%.
将细胞内微电极技术应用于美西螈胃窦黏膜。使用填充有3M氯化钾的15 - 50MΩ微电极获得了稳定的细胞内刺入。在将微电极保持在细胞内的同时能够快速更换黏膜灌流液。利用这些技术,我们能够测量细胞膜和旁路途径的电位和电阻。跨上皮电位为 -4.9±1.3mV,以浆膜溶液为参照。顶端细胞膜电位为 -43.9±0.6mV,细胞相对于黏膜溶液呈负性。基底外侧细胞膜电位为 -48.8±1.3mV,细胞相对于浆膜溶液呈负性。跨上皮电阻为427±66Ω·cm²。顶端与基底外侧膜电阻之比为3.4±0.3。通过在用氨氯吡咪阻断顶端膜钠通透性之前和之后测量总跨上皮电阻以及顶端与基底外侧细胞膜电阻之比来确定跨细胞和细胞旁途径的电阻。顶端细胞膜、基底外侧细胞膜和旁路的电阻分别为2203±585、1296±384和604±81Ω·cm²(平均值±标准误)。根据这些测量结果计算表明,旁路对跨上皮电导的贡献约为85%。