Identification of protein kinase C isoforms in rat mesenteric small arteries and their possible role in agonist-induced contraction.
We have identified immunologically the protein kinase C (PKC) isoforms present in rat mesenteric small arteries, defined their distribution between particulate and soluble fractions, and studied their involvement in phorbol ester-induced contraction. Our analysis revealed the presence of the CA(2+)-dependent PKCs (alpha and gamma), Ca(2+)-independent PKCs (delta and epsilon), and the atypical isoform (zeta). PKCbeta could not be detected, whereas PKCgamma is likely to be of neural origin. All isoforms exhibited different distributions. PKCalpha, PKCepsilon, and PKCzeta were found in both particulate and soluble fractions. In contrast, PKCdelta was mainly in the particulate fraction, and PKCgamma was in the soluble fraction. Phorbol esters, which activate PKC and cause smooth muscle contraction, downregulated only the alpha and delta isoforms. This was associated with a parallel loss of contractile response to phorbol ester. The force developed to submaximal concentrations of noradrenaline was decreased after phorbol dibutyrate pretreatment, although the sensitivity and maximal response were unchanged. Phorbol ester pretreatment did not affect the contractile response to vasopressin. The sensitivity to non-receptor-mediated contraction, caused by k+ in the presence of prazosin, was slightly reduced by 4 alpha- and 4 beta-phorbol ester pretreatment. Maximal tension in response to this agonist was not affected. We conclude that PKCalpha and/or PKCdelta is necessary for phorbol ester-mediated contraction but is not essential for noradrenaline-, vasopressin-, or k(+)-induced contraction, demonstrating differences in the mechanisms involved in the contractile response between these agents.
Ohanian
V
V
Department of Medicine, Manchester, Royal Infirmary, England.
Ohanian
J
J
Shaw
L
L
Scarth
S
S
Parker
P J
PJ
Heagerty
A M
AM
D000818:Animals; D005260:Female; D007527:Isoenzymes / Q000378:metabolism* / Q000502:physiology; D008638:Mesenteric Arteries / Q000187:drug effects / Q000201:enzymology*; D009638:Norepinephrine / Q000494:pharmacology; D015240:Phorbol 12,13-Dibutyrate / Q000494:pharmacology; D011224:Prazosin / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism* / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013347:Subcellular Fractions / Q000201:enzymology; D014018:Tissue Distribution; D014661:Vasoconstriction / Q000187:drug effects / Q000502:physiology*; D014667:Vasopressins / Q000494:pharmacology
D007527:Isoenzymes; D014667:Vasopressins; D015240:Phorbol 12,13-Dibutyrate; D011493:Protein Kinase C; D009638:Norepinephrine; D011224:Prazosin