Correlation Between Myofilament Response to Ca2+ and Altered Dynamics of Contraction and Relaxation in Transgenic Cardiac Cells That Express beta-Tropomyosin

• Published in: Circulation research Vol. 84; no. 7; pp. 745 - 751
• Main Authors: Wolska, Beata M, Keller, Rebecca S, Evans, Christian C, Palmiter, Kimberly A, Phillips, Ronald M, Muthuchamy, Mariappan, Oehlenschlager, James, Wieczorek, David F, de Tombe, Pieter P, Solaro, R. John
• Format: Journal Article
• Language: English
• Published: Hagerstown, MD American Heart Association, Inc 16-04-1999; Lippincott
• Subjects: Actin Cytoskeleton - drug effects; Actin Cytoskeleton - physiology; Adenosine Triphosphatases - metabolism; Animals; Biological and medical sciences; Calcium - pharmacology; Fundamental and applied biological sciences. Psychology; Gene Expression - physiology; Heart; In Vitro Techniques; Mice; Mice, Inbred Strains; Mice, Transgenic; Muscle Contraction - drug effects; Muscle Contraction - physiology; Muscle Fibers, Skeletal - cytology; Muscle Fibers, Skeletal - drug effects; Muscle Fibers, Skeletal - enzymology; Myocardium - cytology; Myocardium - enzymology; Sarcomeres - chemistry; Sarcomeres - enzymology; Tropomyosin - genetics; Tropomyosin - metabolism; Vertebrates: cardiovascular system; Heart; Molecular form; Myofibril; Tropomyosin; Rodentia; Transgenic animal; Gene overexpression; Muscle contraction; Myocyte; Muscular relaxation; Vertebrata; Mammalia; Mouse; Circulatory system; Sarcomere
• ISSN: 0009-7330; 1524-4571

We compared the dynamics of the contraction and relaxation of single myocytes isolated from nontransgenic (NTG) mouse hearts and from transgenic (TG-beta-Tm) mouse hearts that overexpress the skeletal isoform of tropomyosin (Tm). Compared with NTG controls, TG-beta-Tm myocytes showed significantly reduced maximal rates of contraction and relaxation with no change in the extent of shortening. This result indicated that the depression in contraction dynamics determined in TG-beta-Tm isolated hearts is intrinsic to the cells. To further investigate the effect of Tm isoform switching on myofilament activity and regulation, we measured myofilament force and ATPase rate as functions of pCa (-log of [Ca]). Compared with controls, force generated by myofilaments from TG-beta-Tm hearts and myofibrillar ATPase activity were both more sensitive to Ca. However, the shift in pCa50 (half-maximally activating pCa) caused by changing sarcomere length from 1.8 to 2.4 [micro sign]m was not significantly different between NTG and TG-beta-Tm fiber preparations. To test directly whether isoform switching affected the economy of contraction, force versus ATPase rate relationships were measured in detergent-extracted fiber bundles. In both NTG and TG-beta-Tm preparations, force and ATPase rate were linear and identically correlated, which indicated that crossbridge turnover was unaffected by Tm isoform switching. However, detergent extracted fibers from TG-beta-Tm demonstrated significantly less maximum tension and ATPase activity than NTG controls. Our results provide the first evidence that the Tm isoform population modulates the dynamics of contraction and relaxation of single myocytes by a mechanism that does not alter the rate-limiting step of crossbridge detachment. Our results also indicate that differences in sarcomere-length dependence of activation between cardiac and skeletal muscle are not likely due to differences in the isoform population of Tm. (Circ Res. 1999;84:745-751.)