Twenty-four-hour changes in the in vitro serotonin (5-HT) uptake capacity of hypothalamic homogenates and of "Vibratome" slices of the suprachiasmatic nuclear region (SNR) of the hypothalamus were studied in 60-90-day-old Holtzman (albino) rats. Animals acclimated to artificially illuminated (fluorescent, LD 12:12) and temperature controlled (22 +/- 2 C) rooms were killed 6% each of 8 time points. Synaptosomal fractions from homogenates of whole hypothalamus, and slices of the SNR were incubated for 20 min in Krebs-Henseleit buffer with [3H]5-HT. Males showed a single daily peak in SNR uptake at the start of darkness, and a minimum near the onset of light, while a more complex pattern containing 3 peaks and minima occurred in uptake by hypothalamic homogenates. Proestrous females showed a single high amplitude peak SNR uptake during the critical period, just prior to the plasma LH peak determined in the same animals by radioimmunoassay. It is suggested that this short-term and 4-fold increase in SNR uptake of 5-HT may serve to limit free 5-HT and its inhibitory or other effects on the gonadotropin release hormone system and thereby on LH release and ovulation.
Eight cycling female rats were implanted with push-pull cannulae over the region of the suprachiasmatic nuclei (SCN) and allowed 7–10 days for recovery. Perfusion of the SCN continued in these freely behaving rats for 5–6 h of the light period and the subjective scotophase. The release of 5-hydroxyindoleacetic acid (5-HIAA) ranged from 10 to 350 pg 5-HIAA/min. Significantly, the amplitude and characteristics of the output of 5-HIAA were highly location dependent in that rostral cannulae placements revealed high amplitude changes with initial mean values of 40 pg 5-HIAA/min, which increased toward the dark phase to mean peak values of 195 pg 5-HIAA/min. Caudal cannulae placements revealed a low amplitude, high frequency, basal type of 5-HIAA release which did not increase toward the dark period (approximately 5 pg/min). 5-Hydroxytryptophan infusion resulted in a significant marked increase in the basal release of 5-HIAA confirming the biochemical viability of the area undergoing perfusion. These results suggest that the in vivo measurement of 5-HIAA from 5-hydroxytryptamine (5-HT) terminals in the region of the SCN could reflect discrete functional activity of serotonergic terminals within specific regions of the SCN in a freely behaving rat. Furthermore, the biochemical viability of these 5-HT terminals and the ability of the rat’s SCN to exhibit marked differential changes in 5-HT activity emphasizes the physiological relevance of this model system to study neuroendocrine events in freely behaving animals.
The role of hypothalamic and midbrain serotonergic systems in the control of ovulation in the rat has been investigated using 5,7-dihydroxytryptamine (5,7-DHT) lesioning in conjunction with desmethylimipramine pretreatment. Intracisternal injection of 5,7-DHT (100 or 200 microgram) produced a dose-dependent decrease in the incidence of ovulation induced by PMS gonadotrophin (PMSG; 8 IU sc on day 30). Intraventricular injection of 200 microgram 5.7-DHT via the lateral ventricles completely blocked PMSG-induced ovulation. After injection into either site, nonovulatory animals were in proestrus on day 33 and contained fluid-filled uteri. Intracerebral injection of 5,7-DHT into the dorsal or median raphe significantly decreased the numbers of animals induced to ovulate. The extent and specificity of these chemical lesions were evaluated in the suprachiasmatic (SNR) and arcuate-median eminence regions of PMSG-treated rats using an in vitro uptake model. A general feature of every case of inhibited ovulation was the significant decrease in uptake of serotonin in the SNR, indicating destruction of serotonergic inputs to this region. This suggests that serotonergic SNR input from the dorsal raphe region is essential to ovulation. Median raphe lesions appeared to be more extensive than dorsal raphe lesions, involving serotonergic projections to the arcuate-median eminence region. In addition, ascending noradrenergic projections to the SNR were significantly destroyed, also implicating these systems in ovulation control.
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