PKC expression is regulated by dietary K intake and mediates internalization of SK channels in the CCD.
We have used Western blot analysis and immunocytochemistry to determine the effect of dietary K intake on the expression of protein kinase C (PKC) isoforms in the kidney. Western blot has demonstrated that conventional PKC isoforms (alpha and beta), novel PKC isoforms (delta, epsilon, and eta), and atypical PKC isoforms (zeta) are expressed in the renal cortex and outer medulla. Moreover, a low K intake significantly increases the expression of PKC-epsilon in the renal cortex and outer medulla but does not change the expression of PKC-alpha, PKC-beta, PKC-delta, PKC-eta, and PKC-zeta. Also, immunocytochemistry shows that PKC-epsilon isoform is expressed in the cortical collecting duct (CCD) and outer medullary collecting duct (OMCD) and that the intensity of PKC-epsilon staining is higher in the kidney from rats on a K-deficient diet than those on a control diet. Also, we used the patch-clamp technique to study the role of PKC in mediating internalization of ROMK (Kir 1.1)-like small-conductance K (SK) channels induced by phenylarsine oxide (PAO), an agent that inhibits protein tyrosine phosphatase and has been shown to stimulate the internalization of the SK channel in the CCD (Sterling H, Lin DH, Qu RM, Dong K, Herbert SC, and Wang WH. J Biol Chem 277: 4317-4323, 2002). Inhibition of PKC with calphostin C and GF-109203x had no significant effect on channel activity but abolished the inhibitory effect of PAO on SK channels. In conclusion, a low K intake increases the expression of PKC-epsilon isoform in the renal cortex and outer medulla, and PKC is involved in mediating the internalization of SK channels in the CCD induced by stimulation of protein tyrosine kinase activity.
Sterling
Hyacinth
H
Dept. of Pharmacology, BSB Rm. 537, New York Medical College, Valhalla, NY 10595, USA. wenhui_wang@nymc.edu
Lin
Dao-Hong
DH
Chen
Yu-Jung
YJ
Wei
Yuan
Y
Wang
Zhi-Jian
ZJ
Lai
Jian
J
Wang
Wen-Hui
WH
D000465:Algorithms; D000818:Animals; D015153:Blotting, Western; D005260:Female; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000096:biosynthesis; D007672:Kidney Cortex / Q000187:drug effects / Q000378:metabolism*; D007685:Kidney Tubules, Collecting / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D018408:Patch-Clamp Techniques; D015221:Potassium Channels / Q000187:drug effects / Q000378:metabolism*; D024681:Potassium Channels, Calcium-Activated; D017419:Potassium, Dietary / Q000494:pharmacology*; D011493:Protein Kinase C / Q000096:biosynthesis*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D051640:Small-Conductance Calcium-Activated Potassium Channels
D007527:Isoenzymes; D015221:Potassium Channels; D024681:Potassium Channels, Calcium-Activated; D017419:Potassium, Dietary; D051640:Small-Conductance Calcium-Activated Potassium Channels; D011505:Protein-Tyrosine Kinases; D011493:Protein Kinase C