Dissociation of force from myofibrillar MgATPase and stiffness at short sarcomere lengths in rat and toad skeletal muscle

Abstract

1. Single fast-twitch fibres from the extensor digitorum longus muscle of the rat, Rattus norvegicus, and single twitch fibres from the iliofibularis muscle of the cane toad, Bufo marinus, were mechanically skinned and then used to measure maximally Ca2+-activated [( Ca2+] greater than 0.03 mmol l-1) isometric force production, myofibrillar MgATPase activity and fibre stiffness at different sarcomere lengths. MgATP hydrolysis was linked by an enzyme cascade to the oxidation of NADH (nicotinamide adenine dinucleotide, reduced form) and was monitored by a microfluorimetric system. Fibre stiffness was measured from the amplitude of force oscillations generated by small sinusoidal length changes. 2. At sarcomere lengths which were optimal for isometric force production (around 2.7 microns for rat and 2.2 microns for toad fibres) the myofibrillar MgATPase activity (mean +/- S.E.M.) at 21-22 degrees C was found to be 3.80 +/- 0.53 molecules MgATP hydrolysed s-1 per myosin head for eight rat fibres and 6.35 +/- 0.77 s-1 per myosin head for four toad fibres. 3. At sarcomere lengths shorter than 2.7 microns in rat fibres and 2.2 microns in toad fibres, MgATPase and stiffness remained elevated and close to their respective values at 2.7 microns in rat fibres and 2.2 microns in toad fibres even when the isometric force decreased to near zero levels. 4. The dissociation at short sarcomere lengths of myofibrillar MgATPase activity and fibre stiffness from isometric force suggests that the cross-bridge cycle is not greatly affected by double actin filament overlap with the myosin filaments at short sarcomere lengths. Moreover, the results suggest that cross-bridges can be formed by myosin with actin filaments projecting from the nearest Z-line and from the Z-line in the other half of the sarcomere. 5. These results help to reconcile energetic and mechanical data obtained by others at short sarcomere lengths and can be explained within the framework of the sliding filament theory.