Open AccessEffect of Phorbol Esters on Cytosolic Ca2+ Level, Myosin Phosphorylation and Muscle Tension in High K+-Stimulated Bovine Tracheal Smooth Muscle.
Author(s) -
Masaomi Tajimi,
Masaru Hori,
Hiroshi Ozaki,
Hideaki Karaki
Publication year - 1997
Publication title -
japanese journal of pharmacology/japanese journal of pharmacology
Language(s) - English
Resource type - Journals
eISSN - 1347-3506
pISSN - 0021-5198
DOI - 10.1254/jjp.74.195
Subject(s) - myosin , muscle tension , cytosol , phosphorylation , chemistry , myosin light chain kinase , phorbol ester , protein kinase c , smooth muscle , muscle contraction , phorbol , microbiology and biotechnology , medicine , endocrinology , anatomy , biology , biochemistry , enzyme , physical medicine and rehabilitation
To determine the role of protein kinase C (PKC) in bovine tracheal smooth muscle contractility, we examined the effects of phorbol esters on cytosolic Ca2+ level ([Ca2+]i), myosin light chain (MLC) phosphorylation and contractile force in intact muscle and contraction in a permeabilized preparation. In intact muscle, 12-deoxyphorbol 13-isobutyrate (DPB, 1 microM) increased the force without changing [Ca2+]i. High K+ (72.7 mM) induced sustained contraction with sustained increase in [Ca2+]i. In the muscle stimulated by high K+, 50 nM DPB increased the contractile force without changing [Ca2+]i, and 1 microM DPB increased the contractile force with decreasing [Ca2+]i. Thus DPB shifted the [Ca2+]i/force relationship for high K+ to the lower [Ca2+]i in a concentration-dependent manner. In permeabilized muscle, DPB did not induce contraction in the absence of Ca2+ (< < 0 nM), but shifted the Ca2+/force relationship to the lower Ca2+ levels. In the muscle stimulated with high K+, DPB (50 nM and 1 microM) increased MLC phosphorylation and force without changing the MLC phosphorylation/force relationship. DPB (1 microM) increased PKC activity estimated by the translocation from the cytoplasm to the membrane. These results suggest that DPB increases the Ca2+ sensitivity of MLC phosphorylation via the activation of PKC. Furthermore, DPB at higher concentration has an inhibitory effect on stimulated [Ca2+]i.