The pineal gland in mammals is an endocrine organ and generally does not exhibit neuronal characteristics. However, it is known that under culture conditions, cells from newborn rat pineals express properties characteristic of photoreceptors. Here, we studied the potential of rat pineal cells to differentiate into neuronal cell types using different neural markers. Three phenotype markers characteristic of nerve cells, i.e., intense GABA, neuron-specific antigen (HPC-1) and microtubule-associated protein 2 (MAP2) immunoreactivities, were detected in the pineal culture of newborn rats. Expression of the respective neuronal phenotypes appears to be controlled by different mechanisms; in the normal culture medium containing 5.4 mM KCl, numerous cells were stained intensely with anti-GABA antiserum, whereas only a few were stained intensely either with HPC-1 or MAP2 antibody. In a culture medium with a high concentration of KCl (35 mM), which may induce depolarization of nerve cells, numerous cells became strongly positive for HPC-1 or MAP2; both the cell bodies and the neuritic fibers were stained positively. Since cells intensely immunoreactive to GABA, HPC-1 or MAP2 were not found in intact pineals of the rat, the present results indicate that the neuronal potency of the rat pineal cells is expressed only in vitro and is suppressed in vivo, and that the potency is lost during postnatal development. Norepinephrine at 1 microM, which suppresses differentiation of rhodopsin immunoreactive cells, was ineffective in inducing phenotypic expression of neuronal properties in the present system, indicating that the mechanism of suppression of neuronal properties in the intact pineal may be different from the one for photoreceptors.