Superfusion and static culture techniques for measurement of rapid changes in prolactin secretion

Abstract

This chapter describes a superfusion method for the cultured monolayers of anterior pituitary cells that permits the analysis of interactions among various regulatory factors, allows the monitoring of the temporal patterns of hormone release, and is compatible with the simultaneous determination of biochemical and structural changes correlated with secretion. The secretion of prolactin (PRL) is under the regulation of both inhibitory and stimulatory hypothalamic hormones. Dopamine (DA), a well characterized inhibitory hormone, tonically suppresses PRL secretion. The multifactorial regulation of PRL secretion represents a specific but not unique process, in which the sequence and duration of exposure to multiple hypothalamic hormones determine the secretory response. The culture/superfusion technique that is described for dissociated cells permits the unraveling of the hierarchy of the hypothalamic regulation of the secretion of anterior pituitary hormones. Both rapid changes in hormone secretion and cellular events––that is, events occurring within minutes––and slower modulatory effects––that is, effects taking days––can be correlated by the coupling of perifusion techniques with those of cell culture.