Carbachol activates a K + channel of very small conductance in the basolateral membrane of rat pancreatic acinar cells (original) (raw)

1999, Pfl�gers Archiv European Journal of Physiology

Secretion of Clrequires the presence of a K + conductance to hyperpolarize the cell, and to provide the driving force for Clexit via luminal Clchannels. In the exocrine pancreas Clsecretion is mediated by an increase in cytosolic Ca 2+ ([Ca 2+ ] i). Two types of Ca 2+-activated K + channels could be shown in pancreatic acinar cells of different species. However, there are no data on Ca 2+-activated K + channels in rat pancreatic acini. Here we examine the basolateral K + conductance of freshly isolated rat pancreatic acinar cells in cell-attached and cell-excised patch-clamp experiments. Addition of carbachol (CCH, 1 µmol/l) to the bath led to the activation of very small conductance K + channels in cell-attached patches (n=27), producing a noisy macroscopic outward current. The respective outward conductance increased significantly by a factor of 2.1±0.1 (n=27). Noise analysis revealed a Lorentzian noise component with a corner frequency (f c) of 30.3±3.5 Hz (n=19), consistent with channel activity in these patches. The estimated singlechannel conductance was 1.5±0.4 pS (n=19). In cell-excised patches (inside out) from cells previously stimulated with CCH, channel activity was only observed in the presence of K + in the bath solution. Under these conditions f c was 47.6±11.9 Hz (estimated single-channel conductance 1.1±0.2 pS, n=20). The current/voltage relationship of the noise showed weak inward rectification and the reversal potential shifted towards E K + when Na + in the bath was replaced by K +. Channel activity in cellexcised patches was slightly reduced by 10 mmol/l Ba 2+ (23.6±2.1% of the total outward current) and was completely absent when K + in the bath was replaced by Na +. Reduction of the [Ca 2+ ] i from 1 mmol/l to 1 µmol/l in cell-excised experiments decreased the current by 52.3±12.3% (n=5). Expression of K V LQT1, one of the possible candidates for a small-conductance K + channel in rat pancreatic acinar cells, was shown by reverse transcriptase polymerase chain reaction (RT-PCR). In fact, a K V LQT-blocker (chromanol 293B) reduced channel activity in seven excised patches. These data suggest that CCH activates very small conductance K + channels in rat pancreatic acinar cells, most likely via an increase in [Ca 2+ ] i .

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