Phorbol ester-induced potentiation of myogenic tone is not associated with increases in Ca2+ influx, myoplasmic free Ca2+ concentration, or 20-kDa myosin light chain phosphorylation.
We studied the extent of Ca2+ influx, myoplasmic free Ca2+ concentration changes, and phosphorylation of the regulatory 20-kDa myosin light chain (LC20) associated with the potentiation of stretch-induced myogenic tone in the rabbit facial vein. The protein kinase C (PKC) activator, phorbol 12-myristate 13-acetate (PMA), was used to augment Ca(2+)-dependent stretch-induced myogenic tone. Veins stretched to an optimal resting tension in physiological salt solution (PSS) containing 0.4 mM of Ca2+ developed stretch-induced myogenic tone. Tissues incubated in Ca(2+)-free PSS, either with or without PMA (0.1 microM) did not develop myogenic tone. Readmission of Ca2+ (0.4 mM) caused a three-fold increase in the contraction in PMA-treated segments (710 +/- 60 mg, n = 29 v control: 188 +/- 10 mg, n = 24). This increased contraction was not associated with additional increases in either Ca2+ influx (73.5 +/- 6.9 pmol/mg of tissue/min, n = 29 v control: 61.1 +/- 5.7 pmol/mg of tissue/min, n = 24), myoplasmic free Ca2+ concentration or LC20, (0.44 +/- 0.02 mol PO4/mol LC20, n = 9 v control: 0.43 +/- 0.03 mol PO4/mol LC20, n = 7). Our results suggest that PKC activation amplifies stretch-induced myogenic tone in the rabbit facial vein through target proteins that are not associated with regulation of Ca2+ influx, myoplasmic free Ca2+ concentration, or LC20 phosphorylation. We conclude that the PKC-mediated potentiation of stretch-induced myogenic tone is due to an increased sensitivity of the contractile apparatus to Ca2+.
Department of Pharmacology, College of Medicine, University of Vermont, Burlington 05405-0068.
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R
Laporte
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JR
Haeberle
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I
Laher
D000818:Animals; D001692:Biological Transport; D002118:Calcium / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D008297:Male; D009119:Muscle Contraction / Q000187:drug effects; D009124:Muscle Proteins / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000187:drug effects* / Q000378:metabolism; D009219:Myosin-Light-Chain Kinase / Q000378:metabolism*; D009218:Myosins / Q000378:metabolism*; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D011817:Rabbits; D015398:Signal Transduction / Q000187:drug effects*; D013314:Stress, Mechanical; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014680:Veins
D009124:Muscle Proteins; D011493:Protein Kinase C; D009219:Myosin-Light-Chain Kinase; D009218:Myosins; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium