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In the transitional zone of the rat anterior pituitary, spontaneous and LHRH-induced Ca2+ dynamics were visualized using fluo-4 fluorescence Ca2+ imaging. A majority of cells exhibited spontaneous Ca2+ transients, while small populations of cells remained quiescent. Approximately 70% of spontaneously active cells generated fast, oscillatory Ca2+ transients that were inhibited by cyclopiazonic acid (10 μm) but not nicardipine (1 μm), suggesting that Ca2+ handling by endoplasmic reticulum, but not Ca2+ influx through voltage-dependent L-type Ca2+ channels, plays a fundamental role in their generation. In the adult rat anterior pituitary, LHRH (100 μg/ml) caused a transient increase in the Ca2+ level in a majority of preparations taken from the morning group rats killed between 0930 h and 1030 h. However, the second application of LHRH invariably failed to elevate Ca2+ levels, suggesting that the long-lasting refractoriness to LHRH stimulation was developed upon the first challenge of LHRH. In contrast, LHRH had no effect in most preparations taken from the afternoon group rats euthanized between 1200 h and 1400 h. In the neonatal rat anterior pituitary, LHRH caused a suppression of spontaneous Ca2+ transients. Strikingly, the second application of LHRH was capable of reproducing the suppression of Ca2+ signals, indicating that the refractoriness to LHRH had not been established in neonatal rats. These results suggest that responsiveness to LHRH has a long-term refractoriness in adult rats, and that the physiological LHRH surge may be clocked in the morning. Moreover, LHRH-induced excitation and associated refractoriness appear to be incomplete in neonatal rats and may be acquired during development.

endocrinology

Intercellular Communication within the Rat Anterior Pituitary Gland. XV. Properties of Spontaneous and LHRH-Induced Ca2+ Transients in the Transitional Zone of the Rat Anterior Pituitary in Situ