Mechanisms underlying regulation of a barium-sensitive K+ conductance by ATP in single proximal tubule cells isolated from frog kidney.
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 (G(Ba)) in proximal tubule cells isolated from frog kidneys. In this paper we examine the regulation of G(Ba) by ATP. G(Ba) was measured in single cells isolated from frog kidney using the whole-cell patch-clamp technique. G(Ba) 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, G(Ba )was not sensitive to cell-volume changes. Hypotonic shock-induced volume regulation was inhibited by barium and quinidine, inhibitors of G(Ba). 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. G(Ba) was also activated by ADP, via a mechanism dependent on the presence of Mg(2+). However, the responses to ADP and ATP were not additive, suggesting that these nucleotides may share a common mechanism of activation. The regulation of G(Ba) 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 G(Ba) 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 G(Ba) could play a role in volume regulation.
Robson
L
L
Department of Biomedical Science, Alfred Denny Building, University of Sheffield, Sheffield S10 2TN, UK. l.robson@sheffield.ac.uk
Hunter
M
M
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001464:Barium / Q000494:pharmacology; D004553:Electric Conductivity; D015640:Ion Channel Gating / Q000187:drug effects / Q000502:physiology*; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000502:physiology*; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D018408:Patch-Clamp Techniques; D015221:Potassium Channels / Q000378:metabolism*; D011896:Rana temporaria
D015221:Potassium Channels; D001464:Barium; D000255:Adenosine Triphosphate