The stiffness of frog skinned muscle fibres at altered lateral filament spacing.

When the surface membrane is removed from a frog muscle fibre the myofibrils swell, so that the spacing between the filaments increases by 10-30%. In this study, the stiffness of skinned fibres was measured when the lateral spacing of the filament lattice was reduced osmotically using polyvinyl pyrrolidone (PVP), a synthetic linear polymer. In the absence of PVP, the apparent stiffness of relaxed skinned fibres measured during ramp stretches was about 0.05-0.1 of the stiffness of intact fibres. The stiffness increased when the lattice spacing was decreased. The mechanical characteristics of resting stiffness in the presence of PVP (or bovine serum albumin) were similar to those of the short-range elastic component of resting intact fibres. However, when the spacing of skinned fibres was reduced osmotically to that of the intact lattice, the stiffness of skinned fibres was about 1.6 times higher. In the absence of PVP, Ca2+-activated skinned fibres were less stiff than are fully active intact fibres during isometric tetani. The skinned fibre force-extension relation was markedly curved. As the filament spacing was reduced, the stiffness of the Ca2+-activated skinned fibres increased, with little change in isometric tension, and the force-extension curve became more linear. Experiments at varied filament overlap and during feed-back control of sarcomere length, monitored by laser diffraction, showed that PVP increased the stiffness of the sarcomeres. The increase of active stiffness in the presence of PVP could be partially dissociated from the increase of resting stiffness by inclusion of Mg tripolyphosphate in the bathing solutions. It is concluded that the low stiffness and the non-linearity of the force-extension curve observed in fully activated skinned fibres are due primarily to the increase of filament separation that occurs when a fibre is skinned. The mechanism may be related to an increased angle between the subfragment-2 part of the cross-bridge and the backbone of the thick filament, perhaps leading to buckling of cross-bridges under compression.