The 5′-adenosine monophosphate-activated serine/threonine protein kinase (AMPK) is stimulated by energy depletion, increase in cytosolic Ca
2+ activity, oxidative stress, and nitric oxide. AMPK participates in the regulation of the epithelial Na
+ channel ENaC and the voltage-gated K
+ channel KCNE1/KCNQ1. It is partially effective by decreasing PIP
2 formation through the PI3K pathway. The present study explored whether AMPK regulates the renal outer medullary K
+ channel ROMK. To this end, cRNA encoding ROMK was injected into
Xenopus oocytes with and without additional injection of constitutively active AMPK
γR70Q (AMPK
α1-HA+AMPK
β1-Flag+AMPKγ1
R70Q), or of inactive AMPK
αK45R (AMPK
α1K45R+AMPK
β1-Flag+AMPK
γ1-HA), and the current determined utilizing two-electrode voltage-clamp and single channel patch clamp. ROMK protein abundance was measured utilizing chemiluminescence in
Xenopus oocytes and western blot in whole kidney tissue. Moreover, renal Na
+ and K
+ excretion were determined in AMPK
α1-deficient mice (
ampk ?/? ) and wild-type mice (
ampk +/+ ) prior to and following an acute K
+ load (111 mM KCl, 30 mM NaHCO
3, 4.7 mM NaCl, and 2.25 g/dl BSA) at a rate of 500 μl/h. As a result, coexpression of AMPK
γR70Q but not of AMPK
αK45R significantly decreased the current in ROMK1-expressing
Xenopus oocytes. Injection of phosphatidylinositol PI
(4,5)P
2 significantly increased the current in ROMK1-expressing
Xenopus oocytes, an effect reversed in the presence of AMPK
γR70Q. Under control conditions, no significant differences between
ampk ?/? and
ampk +/+ mice were observed in glomerular filtration rate (GFR), urinary flow rate, serum aldosterone, plasma Na
+, and K
+ concentrations as well as absolute and fractional Na
+ and K
+ excretion. Following an acute K
+ load, GFR, urinary flow rate, serum aldosterone, plasma Na
+, and K
+ concentration were again similar in both genotypes, but renal absolute and fractional Na
+ and K
+ excretion were higher in
ampk ?/? than in
ampk +/+ mice. According to micropuncture following a K
+ load, delivery of Na
+ to the early distal tubule but not delivery of K
+ to late proximal and early distal tubules was increased in
ampk ?/? mice. The upregulation of renal ROMK1 protein expression by acute K
+ load was more pronounced in
ampk ?/? than in
ampk +/+ mice. In conclusion, AMPK downregulates ROMK, an effect compromising the ability of the kidney to excrete K
+ following an acute K
+ load.
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