Calcium sensitivity of vertebrate skeletal muscle myosin.

The calcium sensitivity of vertebrate skeletal muscle myosin has been investigated. Adenosinetriphosphatase (ATPase) activity was assayed in a reconstituted system composed of either purified rabbit myosin plus actin or myosin plus actin, tropomyosin, and troponin. The calcium sensitivity of actomyosin Mg-ATPase activity was found to be directly affected by the ionic strength of the assay medium. Actomyosin assayed at approximately physiological ionic strength (120 mM KCl) demonstrated calcium sensitivity which varied between 6 and 52%, depending on the myosin preparation and the age of the myosin. Mg-ATPase activity was increased when calcium was present in the assay medium at physiological ionic strength. Conversely, actomyosin Mg-ATPase activity assayed at a lower ionic strength (15 mM KCl) was inhibited by addition of calcium. Addition of tropomyosin and troponin to the assay increased the calcium sensitivity of the system at the physiological ionic strength still further (up to 99% calcium sensitivity) and conferred calcium sensitivity on the system at the lower ionic strength (greater than 90% calcium sensitivity). A correlation also existed between myosin's calcium sensitivity and the phosphorylated state of light chain 2.

Degenerative changes at the neuromuscular junctions of red, white and intermediate muscle fibers. Part 1. Response to short stump nerve section.

Degeneration at the neuromuscular function following cutting the phrenic nerve at the 9th intercostal space differs in red, white and intermediate skeletal muscle fibers. The ultrastructure of the nerve terminal and the muscle fiber between 12 hours and 21 days following denervation suggests that lack of neurotrophic influences results in responses specific for each fiber type. Degeneration of axon ends is rapid and by 2 days axon terminals are missing from the end-plate areas of all 3 fiber types. Schwann cells "engulf" degenerating axon terminals and eventually replace them in the primary clefts. Schwann cells display specific morphological changes directly related to axonal degeneration. In all instances axon terminal degeneration precedes muscle fiber degeneration. Synaptic cleft changes are similar for all types of muscle fibers. Primary cleft structure appears to be dependent upon neurotrophic influence, whereas secondary cleft structure is relatively unaffected by denervation. Initial changes in subsynpatic regions of muscle fibers include focal loss of sarcomere alignment and skewing of the Z lines. By 21 days myofibrillar disorganization appears most severe in white fibers and least in red muscle fibers. The rate and degree of degeneration of the axon terminal and subjacent muscle fiber are different for each of the 3 muscle fiber types.

Degenerative changes at the neuromuscular junctions of red, white, and intermediate muscle fibers. Part 2. Responses to long stump nerve section and colchicine treatment.

The ultrastructure at the neuromuscular junction (NMJ) of red, white, and intermediate skeletal muscle fibers undergoes specific changes following either unilateral severance of the phrenic nerve or unilateral topical treatment of the phrenic nerve with colchicine. Both procedures were performed in the cervical region and produce similar rates of muscle fiber degeneration. The severity of degeneration appears to be related to muscle fiber type with white fibers being most severely affected and red fibers least affected. Degeneration rates of the axon terminal also correlate with fiber type in the orderwhite, intermediate, red. However, the rates of degeneration of the specific axon terminals are more rapid with surgical severance than with colchicine treatment. Statistical analysis of morphometric data indicates that hemidiaphragms denervated surgically exhibited significant axon terminal degeneration before significant muscle degeneration. Conversely, diaphragmatic muscle fibers of colchicine-treated phrenic nerves exhibit significant degeneration before loss of the axon terminal. Despite reversal of the temporal sequences for loss of axonal and muscular components between the two preparations, degenerative characteristics of muscle fiber structure are similar. This suggests that the presence of fiber-specific neurotrophic substances transmitted from the neuronal cell body to the axon terminal and released at the NMJ may be an important factor in the maintenance of normal muscle fiber morphology.