Exchangeability of actin in cardiac myocytes and fibroblasts as determined by fluorescence photobleaching recovery.
Rhodamine (Rho)-labeled muscle and non-muscle actins were microinjected into cultured embryonic chicken cardiac myocytes and fibroblasts. After incorporation of the fluorescent actin analog into cellular structures, small areas of labeled structures were photobleached with a laser pulse, and fluorescence recovery (FR) was measured to determine the exchangeability of isoactins in these structures. With both Rho-muscle and Rho-non-muscle actins, the FR rate in any part of stress fibers was consistently faster than that observed in any part of myofibrils. Thus, although non-striated (proximal and terminal) portions of nascent myofibrils are similar in appearance and composition to stress fibers, our data clearly revealed differences in actin stability between these two structures. Further, although cardiomyocytes were incapable of discriminating between the incorporation of muscle and non-muscle actin isoforms into myofibrils, FR after photobleaching of Rho-muscle actin was faster than that of Rho-non-muscle actin in immature non-striated portions. This indicates that actin molecules in cardiac myofibrils cannot be readily exchanged by heterotypic non-muscle actin. Fluorescently labeled actin incorporated into non-striated (proximal and terminal) portions of myofibrils and terminal portions of stress fibers was found to be more stable than alpha-actinin. The relative stability of actin could facilitate the formation of nascent Z-bands of myofibrils and the reorganization of stress fibers at these portions.
Suzuki
H
H
Department of Anatomy/Cell Biology, School of Medicine, Chiba University, Japan.
Komiyama
M
M
Konno
A
A
Shimada
Y
Y
D000185:Actinin / Q000032:analysis; D000199:Actins / Q000032:analysis* / Q000378:metabolism* / Q000494:pharmacology; D000209:Acute-Phase Proteins / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D002642:Chick Embryo; D005347:Fibroblasts / Q000737:chemistry; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D008845:Microinjections; D008856:Microscopy, Fluorescence; D018485:Muscle Fibers, Skeletal / Q000737:chemistry* / Q000378:metabolism*; D009206:Myocardium / Q000166:cytology* / Q000378:metabolism; D009210:Myofibrils / Q000737:chemistry / Q000378:metabolism; D010590:Phalloidine; D013314:Stress, Mechanical
D000199:Actins; D000209:Acute-Phase Proteins; D005456:Fluorescent Dyes; C010781:acute-phase protein rho; D000185:Actinin; D010590:Phalloidine; D016650:Fluorescein-5-isothiocyanate