Abstract
K+ channels play an important role in pump-leak coupling and volume regulation in the renal proximal tubule. Previous experiments have identified a barium-sensitive K+ conductance (GBa) in proximal tubule cells isolated from frog kidneys. In this paper we examine the regulation of GBa by ATP. GBa was measured in single cells isolated from frog kidney using the whole-cell patch-clamp technique. GBa was activated by 2 mM intracellular ATP. This activation was enhanced by inhibition of protein kinase C and attenuated by inhibition of protein kinase A, indicating reciprocal regulation by these kinases. Activation by ATP was reduced in the presence of a hypertonic bath solution, suggesting that cell swelling was required. However, after activation to steady-state, GBa was not sensitive to cell-volume changes. Hypotonic shock-induced volume regulation was inhibited by barium and quinidine, inhibitors of GBa. The effect of maximal inhibitory concentrations of barium and quinidine on volume regulation was similar and addition of both blockers together did not augment the inhibitory response. GBa was also activated by ADP, via a mechanism dependent on the presence of Mg2+. However, the responses to ADP and ATP were not additive, suggesting that these nucleotides may share a common mechanism of activation. The regulation of GBa by ATP was biphasic, with a half-maximal activating concentration of 0.89 mM and a half maximal inhibitory concentration of 6.71 mM. The sensitivity to nucleotides suggests that GBa may be regulated by the metabolic state of the cell. Furthermore, the sensitivity to solution osmolality, coupled with the blocker profile of inhibition of volume regulation, suggests that GBa could play a role in volume regulation.
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This work was supported by the Wellcome Trust. Thanks also to J. Hartley for her technical assistance.
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Robson, L., Hunter, M. Mechanisms Underlying Regulation of a Barium -sensitive K+ Conductance by ATP in Single Proximal Tubule Cells Isolated from Frog Kidney. J Membrane Biol 204, 39–47 (2005). https://doi.org/10.1007/s00232-005-0745-8
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DOI: https://doi.org/10.1007/s00232-005-0745-8