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Temperature dependence of active tension in mammalian (rabbit psoas) muscle fibres: effect of inorganic phosphate

Moira E. Coupland, Emma Puchert * and K. W. Ranatunga

1. The effect of added inorganic phosphate (P_i, range 3-25 mM) on active tension was examined at a range of temperatures (5-30 °C) in chemically skinned (0.5 % Brij) rabbit psoas muscle fibres. Three types of experiments were carried out.
2. In one type of experiment, a muscle fibre was maximally activated at low temperature (5 °C) and its tension change was recorded during stepwise heating to high temperature in ~60 s. As found in previous studies, the tension increased with temperature and the normalised tension-(reciprocal) temperature relation was sigmoidal, with a half-maximal tension at 8 °C. In the presence of 25 mM added P_i, the temperature for half-maximal tension of the normalised curve was ~5 °C higher than in the control. The difference in the slope was small.
3. In a second type of experiment, the tension increment during a large temperature jump (from 5 to 30 °C) was examined during an active contraction. The relative increase of active tension on heating was significantly higher in the presence of 25 mM added P_i (30/5 °C tension ratio of 6-7) than in the control with no added P_i (tension ratio of ~3).
4. In a third type of experiment, the effect on the maximal Ca²⁺-activated tension of different levels of added P_i (3-25 mM) (and P_i mop adequate to reduce contaminating P_i to micromolar levels) was examined at 5, 10, 20 and 30 °C. The tension was depressed with increased [P_i] in a concentration-dependent manner at all temperatures, and the data could be fitted with a hyperbolic relation. The calculated maximal tension depression in excess [P_i] was ~65 % of the control at 5-10 °C, in contrast to a maximal depression of 40 % at 20 °C and 30 % at 30 °C.
5. These experiments indicate that the active tension depression induced by P_i in psoas fibres is temperature sensitive, the depression becoming less marked at high temperatures. A reduced P_i-induced tension depression is qualitatively predicted by a simplified actomyosin ATPase cycle where a pre-phosphate release, force-generation step is enhanced by temperature.

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