Isolation of myotoxic component from rattlesnake (Crotalus viridis viridis) venom. Electron microscopic analysis of muscle damage.

Abstract

The pathogenesis of myonecrosis induced by a purified component of rattlesnake (Crotalus viridis viridis) venom was studied at the light and electron microscopic levels. Crude venom was fractionated by gel filtration (Sephadex G-50) followed by cation exchange chromatography (Sephadex C-25). Electrophoretic homogeneity of the isolated myotoxin (Fraction II from C-25 column) was demonstrated in isoelectric focusing and disc gel polyacrylamide gel electrophoresis. White mice were injected intramuscularly with 1.5 mug/g of the purified protein in 0.1 ml of physiologic saline. Light microscopic examination of injected muscle revealed a series of degenerative events including partial vacuolation of muscle cells at 6, 12, and 24 hours and complete vacuolation and loss of striations at 48 and 72 hours. Hemorrhage was not observed. At the electron microscopic level the perinuclear space and sarcoplasmic reticulum were dilated in all samples. By 48 and 72 hours the myofibrils lacked striations and the sarcomeres were disorganized. Plasma membranes and T tubules remained intact in all samples. These results correlated well with the myonecrosis induced by crude Crotalus viridis viridis venom except for several important aspects. The pure component altered skeletal muscle cells specifically, with the sarcoplasmic reticulum being the primary site of action.