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Tension redevelopment following a slack�Crestretch manoeuvre was fitted by a single exponential equation: (1) To examine myofibrillar substrates of PKA, myofibrillar samples from control and propranolol treated rats were incubated with the catalytic subunit of PKA in the presence of radiolabelled ATP, separated by SDS-PAGE, and visualized by autoradiography. Briefly, 10 ��g of skinned cardiac myocytes were incubated with the catalytic subunit of PKA (2.5 U ��l?1) and 50 ��Ci [��-32P]ATP for 30 min. As a control experiment, cardiac myocytes also were incubated with PKA in the presence of PKI (4.4 U ��l?1). The reaction was stopped by the addition of electrophoresis sample buffer and heating at 95��C for 3 min. The samples were then separated by SDS-PAGE (12% acrylamide), silver stained, dried and subsequently exposed to X-ray film for ?24 h at ?70��C. The characteristics of the fast-twitch skeletal muscle fibres, slow-twitch skeletal muscle fibres, and cardiac myocyte preparations are shown in Table 1. Slopes of length�Ctension relationships were determined by linear regression and compared between muscle cell types by one-way ANOVA. A Student�CNeuman�CKeuls post hoc test assessed differences between means. Slopes before and after PKA treatment were compared by Student's t test for paired data. All values are means ��s.d. unless otherwise indicated. P was expected based upon previous measures of length�Ctension relationships at submaximal Ca2+ activations (Allen & Moss, 1987) and length dependence of calcium sensitivity of force (Allen & Moss, 1987; McDonald et al. 1997; Konhilas et al. 2002). Cardiac myocyte preparations, on the other hand, exhibited two populations of length�Ctension relationships, one steeper even than fast-twitch fibres and the other similar to slow-twitch fibres (Fig. 2). The active sarcomere length�Ctension relationships were unrelated to passive force levels as cardiac myocytes exhibited just one population of sarcomere length�Cpassive tension relations, which was considerably steeper than both fast-twitch and slow-twitch skeletal muscle fibres (Fig. 2 inset). Specifically, passive tension was 3.5 �� 2.5 kN m?2 at 2.30 �� 0.04 ��m in cardiac myocyte preparations, which was much greater than fast-twitch (1.1 �� 0.8 kN m?2 at 2.70 �� 0.07 ��m) and slow-twitch (1.4 �� 0.5 kN m?2 at 2.70 �� 0.07 ��m) skeletal muscle fibres. Since ��-adrenergic stimulation is known to shift the left ventricular Frank�CStarling relationship upward (i.e.