Muscle-derived factors enhance cholinergic neuronal expression in the chick embryo-II. In culture studies

The effects of muscle-derived factors on the cholinergic and GABAergic neuronal phenotypes were studied in cultures derived from 3-day-old whole chick embryo (E3WE) consisting of proliferating neuroblasts and 8-day-old chick embryo cerebral hemispheres (E8CH) consisting of differentiated neurons. The effects of limb muscle extract (LME) were examined either when added to the culture medium or to the polylysine coating substratum cultures. We also compared the effects of LME derived from 8- to 15-day-old chick embryos. We found that LME added to the medium not only increased choline acetyltransferase activity, a marker for cholinergic neurons throughout the development of neurons in E3WE or E8CH culture, but also delayed the decline in the activity observed in untreated cultures. The marked increase in choline acetyltransferase activity in E8CH cultures grown on substratum-bound LME as compared to those grown in medium-containing LME, suggests that LME factors may be adhesion-promoting substances stimulating neuronal growth. These findings provide evidence that muscle-derived factors may be important in early cholinergic phenotypic expression and support our previous in ovo studies indicating that target-derived factors in limb muscle extract have general cholinotrophic effect.