Fos-like immunoreactivity in cultured rat pinealocytes

The expression of Fos-like proteins in cultured rat pinealocytes was investigated immunocytochemically. Pineal glands of 2-day-old rats were dissociated and maintained in culture for 13 days, the last 24 h deprived of animal sera. The pinealocytes were identified morphologically and by serotonin-immunostaining. Incubations with phorbol dibutyrate (PDBu), 8-bromo-cAMP and isoprenaline were carried out for 2 h to test whether Fos-like protein synthesis is activated via serum response element (SRE), calcium/cyclic AMP response element (Ca/CRE) and beta-receptors, respectively. In untreated serum-deprived control cultures, one third of pinealocytes were Fos-immunoreactive suggesting that Fos protein may have a homeostatic role. In cultures incubated with PDBu, more than 70% of the pinealocytes had Fos-like immunoreactive material in their nuclei. Similarly, in cultures incubated with 8-bromo-cAMP or isoprenaline, more than 70% of the pinealocytes were Fos positive. The c-fos gene could be involved in the regulation of pineal melatonin synthesis and SRE and Ca/CRE probably participate in its activation process in pinealocytes.