Kinetics of functional and morphological changes during decoupling and recoupling induced by glycerol in isolated muscle fibres of the crayfish.

The development of ultrastructural changes in the T-system of isolated muscle fibres of the crayfish by the glycerol procedure is described in correlation with the dissociation of excitation-contraction (E--C) coupling as well as with recoupling of the E--C link. The sequence of events in the process of disconnection of the tubules is as follows: dilation of the T-system tubules, disconnection of the constricted tubular segments from the surface membrane and from the T-system vesicle, disappearance of the lumen and its disintegration. The decoupled state is characterised by the presence of round vesicles uniformly distributed in the entire volume of the fibre. The volume of vesicles accounts well for the residual postglycerol volume increase (15%) of the muscle fibres. Functional and structural recovery can be induced by reapplication of glycerol to fibres decoupled and vesiculated with concentrations of glycerol less than or equal to 300mmol.l(-1) in crayfish saline. The restitution starts with the organisation of the material of the disintegrated connecting segment of the T-system tubule into small vesicles which coalesce to form the tubule from the vesicular site. At the same time the surface membrane is invaginated toward the vesicle, thus forming the tubule from the surface membrane site. Recovery starts already in the first minute after application of glycerol and is completed within approximately 15 min.