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Am J Physiol Heart Circ Physiol (October 7, 2004). doi:10.1152/ajpheart.00407.2004
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Submitted on May 3, 2004
Accepted on October 5, 2004

Impact of {beta}-Myosin Heavy Chain Isoform Expression on Cross-Bridge Cycling Kinetics

Veronica L. Rundell1, Vlasios Manaves1, Anne F. Martin1, and Pieter P. de Tombe1*

1 Department of Physiology and Biophysics, University of Illinois at Chicago, Chicago, IL, USA

* To whom correspondence should be addressed. E-mail: pdetombe{at}uic.edu.

Myosin heavy chain (MyHC) isoforms, {alpha} and {beta}, have intrinsically different ATP hydrolysis activities (ATPase) and therefore cross-bridge cycling rates when in solution. There is considerable evidence of altered MyHC expression in rodent models of cardiac disease, however, the effect of incremental {beta}-MyHC expression over a wide range on the rate of high strain, isometric, cross-bridge cycling is yet to be ascertained. Accordingly, we treated male rats with 6-n-propyl 2-thiouracil (PTU 0.8g/L in drinking water) for short intervals (6, 11, 16 and 21 days) to generate cardiac MyHC patterns in transition from predominantly {alpha}-MyHC to predominantly {beta}-MyHC. Steady state calcium dependent tension development and tension-dependent ATP consumption (tension cost: proportional to cross-bridge cycling) were measured in chemically permeabilized (skinned) right ventricular muscles at 20° C. To assess dynamic cross-bridge cycling kinetics the rate of force redevelopment (ktr) was determined following rapid release-restretch of fully activated muscles. MyHC isoform content in each experimental muscle was measured by SDS-PAGE followed by densitometry. Alpha-MyHC content decreased significantly and progressively with length of PTU treatment, [Days of PTU 6: 68% ± 5; 11: 58% ± 4; 16: 37% ± 4; 21: 27% ± 6; p < 0.001 by ANOVA]. Tension cost decreased, linearly, with decreased {alpha}-MyHC content [Days of PTU 6: 6.7 ± 0.4; 11: 5.6 ± 0.5; 16: 4.0 ± 0.4; 21: 3.9 ± 0.3 ATPase/Tension p < 0.001 by ANOVA]. Likewise, the rate of force redevelopment, ktr, was significantly and progressively depressed with length of PTU treatment [Days of PTU 6: 11.1 ± 0.6; 11: 9.1 ± 0.5; 16: 8.2 ± 0.7; 21: 6.2 ± 0.3 expressed as s-1 p < 0.05 by ANOVA] Thus, cross-bridge cycling, under high strain, for {alpha}-MyHC is 3 times higher than for {beta}-MyHC. Furthermore, under isometric conditions, {alpha}-MyHC and {beta}-MyHC cross-bridges hydrolyze ATP independently of one another.




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