The whole-cell configuration of the patch-clamp technique was used to record K+ currents from type I cells enzymatically dispersed from the neonatal rat carotid body. The current-voltage (I-V) relationship for the K+ currents showed a prominent, outward shoulder at test potentials of between +10 and +30 mV. 2. The shoulder of the I-V curve could be enhanced by raising extracellular Ca2+ concentration or by bath application of 5 microM-Bay K 8644. It could also be suppressed by bath application of 100 microM-Cd2+ or 5 microM-methoxyverapamil (D600), indicating that a large component of the K+ current in these cells was activated by an influx of Ca2+ through its own channels during cell depolarization. 3. Potassium currents were also reversibly suppressed by 8 nM-charybdotoxin but unaffected by 100 nM-apamin, suggesting that the Ca2(+)-dependent K+ current was carried through large or intermediate conductance Ca2(+)-activated K+ channels. 4. Lowering the pH of the bathing medium from 7.40 to 7.00 reversibly reduced the K+ current amplitudes, and suppressed the shoulder normally seen in the I-V relationship. This effect was enhanced in the presence of 5 microM-Bay K 8644 and abolished in the presence of 5 microM-D600. 5. It is concluded that the Ca2(+)-dependent K+ channels of type I carotid body cells are selectively suppressed by extracellular acidity. Possible mechanisms underlying this effect, and its role in excitation of the carotid body are discussed.
摘要
采用膜片钳技术的全细胞模式,记录新生大鼠颈动脉体酶解分散的I型细胞的钾离子电流。钾离子电流的电流-电压(I-V)关系在+10至+30 mV的测试电位下显示出一个明显的外向峰。2. 通过提高细胞外钙离子浓度或浴槽应用5微摩尔- Bay K 8644可增强I-V曲线的峰。浴槽应用100微摩尔- Cd2+或5微摩尔-甲氧基维拉帕米(D600)也可抑制该峰,表明这些细胞中钾离子电流的很大一部分是在细胞去极化过程中通过其自身通道的钙离子内流激活的。3. 8纳摩尔- 蝎毒素也可可逆地抑制钾离子电流,但100纳摩尔- 蜂毒明肽对其无影响,提示钙依赖性钾离子电流是通过大电导或中电导钙激活钾离子通道传导的。4. 将灌流液的pH从7.40降至7.00可可逆地降低钾离子电流幅度,并抑制I-V关系中通常出现的峰。在5微摩尔- Bay K 8644存在时这种效应增强,而在5微摩尔- D600存在时则消除。5. 得出结论,I型颈动脉体细胞的钙依赖性钾离子通道被细胞外酸度选择性抑制。讨论了这种效应的可能机制及其在颈动脉体兴奋中的作用。