Synchronous in situ ATPase activity, mechanics, and Ca2+ sensitivity of human and porcine myocardium.

Griffiths PJ., Isackson H., Pelc R., Redwood CS., Funari SS., Watkins H., Ashley CC.

Flash-frozen myocardium samples provide a valuable means of correlating clinical cardiomyopathies with abnormalities in sarcomeric contractile and biochemical parameters. We examined flash-frozen left-ventricle human cardiomyocyte bundles from healthy donors to determine control parameters for isometric tension (P(o)) development and Ca(2+) sensitivity, while simultaneously measuring actomyosin ATPase activity in situ by a fluorimetric technique. P(o) was 17 kN m(-2) and pCa(50%) was 5.99 (28 degrees C, I = 130 mM). ATPase activity increased linearly with tension to 132 muM s(-1). To determine the influence of flash-freezing, we compared the same parameters in both glycerinated and flash-frozen porcine left-ventricle trabeculae. P(o) in glycerinated porcine myocardium was 25 kN m(-2), and maximum ATPase activity was 183 microM s(-1). In flash-frozen porcine myocardium, P(o) was 16 kN m(-2) and maximum ATPase activity was 207 microM s(-1). pCa(50%) was 5.77 in the glycerinated and 5.83 in the flash-frozen sample. Both passive and active stiffness of flash-frozen porcine myocardium were lower than for glycerinated tissue and similar to the human samples. Although lower stiffness and isometric tension development may indicate flash-freezing impairment of axial force transmission, we cannot exclude variability between samples as the cause. ATPase activity and pCa(50%) were unaffected by flash-freezing. The lower ATPase activity measured in human tissue suggests a slower actomyosin turnover by the contractile proteins.

DOI

10.1016/j.bpj.2009.07.058

Type

Journal article

Journal

Biophys J

Publication Date

04/11/2009

Volume

97

Pages

2503 - 2512

Keywords

Actomyosin, Adenosine Triphosphatases, Adenosine Triphosphate, Animals, Biophysics, Calcium, Fluorometry, Glycerol, Humans, Hydrogen-Ion Concentration, Microscopy, Atomic Force, Myocardial Contraction, Myocardium, Myosins, Swine

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