Formation of acetylcholine receptor clusters in mammalian sternohyoid muscle regenerating in the absence of nerves

Abstract

When the sternohyoid muscle from the rat is grafted, the original muscle fibers, including the membranes at the neuromuscular junction, degenerate irreversibly. New muscle fibers regenerate inside of the basal laminae remaining from the original muscle fibers. In this study rhodamine-[alpha]-bungarotoxin and electron microscopy have been used to demonstrate that acetylcholine receptor (AchR) clusters and synaptic folds are restored to the regenerating myotubes even when innervation to the grafts is prevented. The AchR clusters and synaptic folds colocalized with acetylcholinesterease that persisted at the original synaptic basal lamina. The AchR clusters were not restored if the original innervation band was removed from the muscle at the time of grafting. Lengths of the AchR clusters were measured in animals ranging in weight from 50 to 700 g. The lengths of clusters in the grafts were proportional to the lengths of those in the preoperative controls, suggesting that quantitative morphogenetic information persists through the period of degeneration and regeneration. However, the distribution of the AchRs within the clusters differed slightly from controls. Extrajunctional AchR clusters were present initially, but later disappeared. The sizes of these clusters were unrelated to the sizes of the junctional AchR clusters. This study demonstrates that morphogenetic cues persist within the region of the original motor and plate, possibly associated with the synaptic basal lamina.