Autoradiographic visualization of muscarinic receptors on rat paratracheal neurons in dissociated cell culture.

An autoradiographic method was used to determine the distribution of muscarinic receptors on cells cultured from the trachealis muscle of 12-13-day-old rats. Cells identified in these culture preparations included neurones, fibroblasts, smooth muscle, and glial and epithelial cells. The cultured cells were incubated with the specific, irreversible ligand [3H]propylbenzylylcholine mustard, and the autoradiographs generated showed that most, if not all, of the paratracheal neurones observed in these cultures were specifically labelled. Both the neuronal cell body and associated neurites were evenly labelled over their entire surface. Neither the pattern nor the density of neuronal labelling appeared to be influenced by close association with other cultured cell types. Autoradiographic grains for muscarinic receptors also appeared to be uniformly distributed over smooth muscle cells and epithelial cell groups in culture. In contrast, no specific labelling was associated with cultured fibroblasts, glial cells and other non-neuronal supporting cells. The precise localization of muscarinic receptors on different cell types in culture may prove to be useful knowledge in the design of an effective and specific antimuscarinic bronchodilator.