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The behavior of fetal canine cardiac cells in culture: synthesis and phosphorylation of myosin.
Authors:T Andreasen  J J Castles  W Y Saito  K Chacko  C Fenner  D T Mason  J Wikman Coffelt
Institution:1. Department of Internal Medicine (Sections of Cardiovascular Medicine and Rheumatology), University of California School of Medicine, Davis, California 95616 USA;2. Department of Anatomy, University of California School of Medicine, Davis, California 95616 USA
Abstract:This study describes the first in vitro culturing of canine cardiac cells. Canine cardiac myosin which was synthesized in a 14-day tissue culture, based on l-3H]leucine incorporation, was precipitated with goat γG antimyosin (cardiac-specific) and analyzed on dodecylsulfate gels; the specific activity of the highly purified myosin chains was determined. Incorporation of 32PO4 was similarly analyzed. The comparative degree of synthesis and phosphorylation of myosin chains, occurring in culture, was the same as that obtained in vivo. Both l-3H]leucine and 32PO4 incorporation were inhibited by addition of cycloheximide to the culture medium. Removal of 32PO4 from myosin heavy chains with base treatment indicated the presence of phosphoserine and/or phosphothreonine in canine cardiac myosin heavy chains. Myosins from fetal and adult canine cardiac tissue were immunologically identical with each other and with the cultured fetal tissue; all had similar myosin ATPase activity and the degree of heavy chain phosphorylation was similar. The tissue and techniques used here gave a high yield of cardiac myocytes based principally on synthesis of cardiac-specific myosin.
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