In vitro populations of neurons dissociated from the superior cervical ganglion of the rat embryo can acquire many of the properties of cholinergic neurons and yet retain some of the characteristics of adrenergic neurons. To determine whether one neuron can stably express properties of two transmitter systems, identified neurons were characterized by both immunocytochemical and electrophysiological methods. It was found that more than 90% of the neurons that formed functional cholinergic synapses had tyrosine hydroxylase within their cytoplasm. Furthermore, this mixed phenotypic expression persisted for more than 1 month. It is concluded that, under certain conditions in vitro, most neurons from the superior cervical ganglion have at least the potential for dual function.