Injury and recovery of fast and slow skeletal muscle fibers affected by ACL myotoxin isolated from Agkistrodon contortrix laticinctus (Broad-Banded copperhead) venom.

The response of different types of skeletal muscle fibers to a snake venom PLA2 myotoxin was tested in vivo by injecting ACL myotoxin (ACLMT) into mice. Both the soleus (slow-twitch) and gastrocnemius (fast-twitch) were examined at different time periods (3 h, 3 and 21 d) after the injection. All animals received 5 mg/kg myotoxin into the subcutaneous lateral region of the right hind limb, near the Achilles tendon; contralateral muscles were used as controls. Cross-sections (10 microm) of frozen muscle tissue were cut from the medial region of the muscle. Alternate serial sections were stained either with toluidine blue or for acid phosphatase, myofibrillar ATPase activity after alkali (pH 10.3) or acid preincubation (pH 4.3), succinate dehydrogenase or acetylcholinesterase. Several stages of necrosis were observed 3 h after ACLMT injection, in both superficial and deep regions of both muscles. In these same regions 3 d after injection, clusters of regenerated muscle fibers were present, and some of them presented AChE activity. Twenty-one days after ACLMT injection the muscle fibers of soleus and gastrocnemius presented only chronic signs of damage such as split fibers and centralized nuclei. Using m-ATPase reactions it was possible to determine that both muscle fiber types I and II were injured in both muscles. The number of type IIC fibers was significantly increased, and the number of type II fibers significantly decreased in the gastrocnemius 21 d after ACLMT injection, suggesting a change in muscle fiber type from type II to type I, through type IIC. The increased number of type IIC fibers and the presence of AChE activity in clusters of regenerating fibers and split fibers indicate that injury by ACLMT produces axonal remodeling and muscle fiber type change.