Hypotonic activation of volume-sensitive outwardly rectifying chloride channels in cultured PASMCs is modulated by SGK.
The serum- and glucocorticoid-inducible kinase (SGK) is a serine/threonine protein kinase (PK) transcriptionally regulated by corticoids, serum, and cell volume. SGK regulates cell volume of various cells by effects on Na(+) and K(+) transport through membrane channels. We hypothesized a role for SGK in the activation of volume-sensitive osmolyte and anion channels (VSOACs) in cultured canine pulmonary artery smooth muscle cells (PASMCs). Intracellular dialysis through the patch electrode of recombinant active SGK, but not kinase-dead Delta60-SGK-K127M, heat-inactivated SGK, or active Akt1, partially activated VSOACs under isotonic conditions. Dialysis of active SGK before cell exposure to hypotonic medium significantly accelerated the activation kinetics and increased the maximal density of VSOAC current. Exposure of PASMCs to hypotonic medium (230 mosM) activated phosphatidylinositol 3-kinases (PI3Ks) and their downstream targets Akt/PKB and SGK but not PKC-epsilon. Inhibition of PI3Ks with wortmannin reduced the activation rate and maximal amplitude of VSOACs. Immunoprecipitated ClC-3 channels were phosphorylated by PKC-epsilon but not by SGK in vitro, suggesting that SGK may activate VSOACs indirectly. These data indicate that the PI3K-SGK cascade is activated on hypotonic swelling of PASMCs and, in turn, affects downstream signaling molecules linked to activation of VSOACs.
Department of Pharmacology, University of Nevada School of Medicine, Reno, NV 89557-0270, USA.
Ge-Xin
GX
Wang
Cian
C
McCrudden
Yan-Ping
YP
Dai
Burton
B
Horowitz
Joseph R
JR
Hume
Ilia A
IA
Yamboliev
D000818:Animals; D002478:Cells, Cultured; D018118:Chloride Channels / Q000378:metabolism / Q000502:physiology*; D004285:Dogs; D004553:Electric Conductivity; D004594:Electrophysiology; D007038:Hypotonic Solutions / Q000494:pharmacology*; D017874:Immediate-Early Proteins; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000378:metabolism*; D009687:Nuclear Proteins; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism; D051744:Protein Kinase C-epsilon; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism / Q000502:physiology*; D011518:Proto-Oncogene Proteins / Q000378:metabolism / Q000502:physiology; D051057:Proto-Oncogene Proteins c-akt; D011651:Pulmonary Artery / Q000166:cytology / Q000378:metabolism*; D000099184:Serum-Glucocorticoid Regulated Kinases
D018118:Chloride Channels; D007038:Hypotonic Solutions; D017874:Immediate-Early Proteins; D009687:Nuclear Proteins; D019869:Phosphatidylinositol 3-Kinases; D011493:Protein Kinase C; D051744:Protein Kinase C-epsilon; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D051057:Proto-Oncogene Proteins c-akt; D000099184:Serum-Glucocorticoid Regulated Kinases; C115376:ClC-3 channel