We have used the perforated-patch variation of whole cell patch-clamp techniques, measurements of cytosolic calcium with use of fura 2, and secretion measurements with use of the reverse-hemolytic plaque assay to address the role of depolarizing background currents in maintaining spontaneous action potentials and spontaneous secretion from rat lactotrophs in primary culture. Replacement of bath sodium with tris(hydroxymethyl)aminomethane or N-methyl-D-glucamine caused a dramatic hyperpolarization of the cells, a cessation of spontaneous action potentials, and an increase in input resistance of cells. Tetrodotoxin had no effect on spontaneous action potentials, and removal of bath calcium stopped spiking but did not hyperpolarize the cells. The hyperpolarization in response to removal of bath sodium was associated with a decrease in cytosolic calcium levels. Finally, removal of bath sodium caused a decrease in spontaneous secretion of prolactin from lactotrophs. These data suggest that a background sodium current is essential to drive the membrane to threshold for firing spontaneous calcium-dependent action potentials in lactotrophs. This, in turn, results in elevated intracellular calcium, which supports spontaneous secretion of prolactin from these cells.
Spontaneous action potentials and circumnutation in Helianthus annuus
