We investigated the importance of calcium (Ca2+) influx in increasing the steady state concentrations of mRNAs coding for the pituitary peptides PRL, alpha, and TSH, LH, and FSH beta-subunits. Adult female rat pituitaries were dissociated, plated for 48 h, then inserted into perifusion chambers. Secretory responses were measured after 2 and 22 h of perifusion, and after 24 h, the cells were recovered, total RNA was extracted, and mRNAs were assayed by dot blot hybridization. The first experiment examined the effect of the Ca2+ channel blocker verapamil (100 microM) on the stimulatory action of pulsatile TRH (4 nM; 60-min interval) or GnRH (100 pM; 60-min interval) on pituitary mRNAs. TRH pulses induced a significant increase (49-56%) in PRL, alpha, and TSH beta mRNAs. Similarly, GnRH pulses stimulated a rise in alpha (64%) and FSH beta (50%) mRNAs, but not LH beta. The effects of pulsatile TRH or GnRH were eliminated when verapamil was added to the medium, suggesting that Ca2+ influx is critical to the stimulatory action of TRH or GnRH. The second experiment examined the effect of pulsatile vs. continuous increases in intracellular Ca2+ on pituitary mRNA expression. Pulsatile Ca2+ signals were produced by giving 60-min pulses of 50 mM KCl, Bay K 8644 (10 microM), or Bay K 8644 (10 microM; in the presence of 10 mM KCl in the injectate) and vehicle pulses to controls. Continuous increases in intracellular Ca2+ were induced by perifusion with medium containing the Ca2+ ionophore A23187 (20 microM), and these groups were compared to that receiving continuous verapamil. Pulsatile increases in Ca2+ influx (KCl or Bay K 8644) stimulated significant elevations in all mRNAs studied (36-74% increase vs. controls), with the exception of TSH beta. The magnitude of the mRNA responses to pulsatile Ca2+ (vs. controls) was similar to that observed after TRH and GnRH pulses. In contrast, only LH beta was increased by A23187 (42% increase vs. controls; P < 0.05). PRL and alpha mRNAs were selectively diminished by A23187 (57% and 83% decreases vs. controls, respectively; P < 0.05) and verapamil (67% and 60%; P < 0.05). The data show that expression of these pituitary genes is regulated by Ca2+ and that a pulsatile Ca2+ signal is required to stimulate PRL, alpha, and FSH beta (but not LH beta).(ABSTRACT TRUNCATED AT 400 WORDS)