Park K, Brown P D
School of Biological Sciences, University of Manchester, United Kingdom.
Am J Physiol. 1995 Mar;268(3 Pt 1):C647-50. doi: 10.1152/ajpcell.1995.268.3.C647.
The effects of intracellular pH (pHi) on Ca(2+)-activated Cl- currents in rat lacrimal gland acinar cells were examined. Cl- currents were recorded by conventional whole cell patch-clamp methods using K(+)-free and Na(+)-free solutions. pHi was varied by using electrode solutions with pH at 6.8, 7.3, or 7.8, and Ca2+ activity was buffered at 100 nM with ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid. Increasing pH from 6.8 to 7.8 was found to increase whole cell currents. The currents observed exhibited time-dependent activation at depolarizing potentials and time-dependent inactivation at hyperpolarizing potentials (pH 7.8). This behavior is characteristic of Ca(2+)-activated Cl- channels in lacrimal gland cells. The selectivity of the current was examined at pH 7.8 by removing Cl- from the bath solution. This maneuver caused a positive shift in the reversal potential, as expected for a Cl(-)-selective current. Thus increasing pHi appears to activate Ca(2+)-activated Cl- channels. The possibility that an increase in pHi may help sustain Cl- channel activity during secretory activity is discussed.
研究了细胞内pH值(pHi)对大鼠泪腺腺泡细胞中钙激活氯离子电流的影响。使用无钾和无钠溶液,通过传统的全细胞膜片钳方法记录氯离子电流。通过使用pH值为6.8、7.3或7.8的电极溶液来改变pHi,并使用乙二醇双(β-氨基乙醚)-N,N,N',N'-四乙酸将钙活性缓冲在100 nM。发现将pH从6.8提高到7.8会增加全细胞电流。观察到的电流在去极化电位下表现出时间依赖性激活,在超极化电位下(pH 7.8)表现出时间依赖性失活。这种行为是泪腺细胞中钙激活氯离子通道的特征。在pH 7.8时,通过从浴液中去除氯离子来检查电流的选择性。如预期的氯离子选择性电流一样,这一操作导致反转电位正向偏移。因此,提高pHi似乎会激活钙激活氯离子通道。讨论了pHi升高可能有助于在分泌活动期间维持氯离子通道活性的可能性。