Efflux of 3H-5-hydroxytryptamine from Rat Hypothalamic Slices by Continuous Electrical Stimulation: Frequency-dependent Responses to Serotonergic Antagonists and 5-hydroxytryptamine

Rat hypothalamic slices were incubated with 3H-5-hydroxytryptamine and superfused in the presence of paroxetine to inhibit 5-hydroxytryptamine (5-HT) reuptake. The slices were continuously stimulated electrically with rectangular pulses at varying frequencies. Continuous stimulation for up to 42 min at 1 Hz or at 3 Hz evoked a steady efflux of tritium that slowly decayed with time. The efflux produced by continuous stimulation at 5 Hz declined more rapidly with time. Continuous stimulation at 1 Hz in the presence of increasing concentrations of unlabelled 5-HT produced a concentration-dependent decrease in tritium efflux. The presence of methiothepin (0.5 mumol/l), quipazine (10 mumol/l) and (-)- but not (+)-propranolol (1 mumol/l) attenuated this response to 5-HT. From these data, the apparent pA2 values were calculated and found to be in agreement with published values. Frequency-dependent responses were determined using a "cumulative stimulation" protocol whereby the slices were subjected to three consecutive 14 min periods of stimulation at increasing frequencies (1, 3 and then 5 Hz). Unlabelled 5-HT (1 mumol/l) inhibited electrically-evoked tritium efflux more at 1 than at 5 Hz. Methiothepin (0.5 mumol/l) and quipazine (10 mumol/l) enhanced the stimulated efflux in a manner inversely related to the frequency of stimulation. Neither (+)- nor (-)-propranolol enhanced stimulated tritium efflux at any of the three frequencies tested. It is concluded that continuous electrical stimulation of rat hypothalamic slices at a low frequency provides a rapid means of obtaining apparent affinities and intrinsic activities of drugs that modify the serotonergic autoreceptor.(ABSTRACT TRUNCATED AT 250 WORDS)