Regeneration of muscle axons in the frog is directed by diffusible factors from denervated muscle and nerve tubes.

In the frog, peripheral muscle axons regenerate after a lesion to reinnervate the original synaptic sites on muscle fibers. Previous experiments in the frog have shown that satellite cells of the nerve tube direct the outgrowth of regenerating muscle axons over distances of many millimeters. In the present experiments, denervated muscle was used as a target for regenerating muscle axons. Muscle and satellite cells of the nerve tube also were placed in filters to determine if their influence on axonal outgrowth was exerted by diffusible factors. Filters were used with a pore size of 0.22 micron. With this pore size, target cells were isolated from physical contact with the surrounding cells; yet an exchange of fluids--and therefore of molecules released by the target cells--could occur across the filter. In the presence of denervated muscle or satellite cells of the nerve tube in filters, regenerating axons turn and grow toward the target cells. This influence on the direction of axonal outgrowth was produced over distances of 6 mm by muscles and 4 mm by cells of the nerve tubes. This directed outgrowth is in marked contrast to the random pattern of outgrowth in the absence of the targets. The present findings set the stage for tissue culture experiments in which the phenomena observed in vivo can be analyzed in terms of mechanisms. The present finding that denervated muscle attracts regenerating axons means that sufficient material may be available for the characterization and isolation of the relevant molecules.