The use of in vivo direct drug application to assess neural regulation of muscle properties.

Skeletal muscle is a convenient model system for studying basic questions on the neural regulation of synaptogenesis and of many properties of sarcolemma and contractile apparatus. The study of the neural signals involved in a particular regulation and of the mediating intracellular pathways, requires the chronic application of drugs, second messengers, antibodies, trophic factors and the like. The most common way of application is in vitro treatment of muscle cell lines or primary myotube cultures. As an alternative to tissue culture, we developed a technique for in vivo application of the agents under study directly on skeletal muscle. An initial surgical step secures the tip of a fine polyethylene catheter (≤250 μm) onto the proximal surface of the adult rat soleus muscle. The onset of perfusion of the solution containing the agent, however, starts only 10–15 days later, pushed by an implanted Alzet pump connected to the catheter. After chronic in vivo treatment for the appropriate number of days, the muscle is processed as needed. We used various agents which are known to affect muscle fibre acetylcholine receptor regulation, namely, CGRP, tetrodotoxin, and KCl and obtained results which demonstrate the full effectiveness of this way of application.