5HT and 5HIAA dialysate levels within the arcuate nucleus of the hypothalamus: relationship with photoperiod-driven differences in serum prolactin and luteinizing hormone in the Siberian hamster

This study examined the relationship between dialysate levels of serotonin (5HT), and its major metabolite 5HIIAA within the arcuate nucleus of the hypothalamus (ARC) and serum gonadotropin levels under two different in vivo paradigms. Experiment 1 evaluated the relationship between dialysate levels of 5HT and 5HIAA within the ARC and circulating prolactin (PRL) and lutenizing hormone (LH) levels under long- and short-day photoperiod conditions. In experiment 2, the profile of 5HT and 5HIAA dialysate levels within the ARC on the afternoon of proestrous was investigated to determine if changes in serotonergic neurotransmission are correlated with preovulatory surges in LH and PRL. Adult male and female Siberian hamsters were housed either in long-day (16L:8D) or short-day (10L:14D) photoperiods for 8 weeks. Dialysis samples were collected every hour for 5 h (12.00-17.00 h) and blood samples were collected via a jugular cannula every hour for analysis of LH and PRL levels. ARC 5HT and 5HIAA dialysate levels were significantly higher in short-day exposed female hamsters, correlating with suppressed basal LH and PRL secretion when compared to their long-day counterparts. Short-day housed male hamsters displayed significantly higher dialysate levels of 5HIAA than males exposed to a long-day photoperiod-5HT was below the lower limit of detection regardless of photoperiod exposure. Long-day females in proestrus showed no change in dialysate levels of 5HT or 5HIAA within the ARC just prior to the onset of the afternoon surge of LH and PRL. Our results indicate that elevated 5HT and 5HIAA dialysate levels within the ARC may regulate photoperiod effects upon LH and PRL secretion, but not the preovulatory surges of LH and PRL.     

Effects of an epinephrine synthesis inhibitor, SKF64139, on the secretion of luteinizing hormone in ovariectomized female rats

In order to test the involvement of central epinephrine systems in luteinizing hormone (LH) secretion, the epinephrine synthesis inhibitor, SKF64139, was administered to ovariectomized rats treated with ovarian hormones. This agent significantly decreased resting LH concentrations in ovariectomized, hormonally untreated rats and completely suppressed the stimulatory feedback effects of estrogen plus progesterone. In these animals, hypothalamic concentrations of epinephrine, but not norepinephrine or dopamine, were significantly decreased. It is suggested that brain epinephrine may participate in the neural control of LH secretion in rats.     

Luteinizing hormone and luteinizing hormone-releasing hormone secretion is under locus coeruleus control in female rats

It has been suggested that norepinephrine (NE) from the locus coeruleus (LC) plays an important role in triggering the preovulatory surge of gonadotropins. This work intended to study the role of LC in luteinizing hormone (LH) secretion during the estrous cycle and in ovariectomized rats treated with estradiol and progesterone (OVXE(2)P) and to correlate it with LH releasing hormone (LHRH) content in the medial preoptic area (MPOA) and median eminence (ME). Female rats on each day of the estrous cycle and OVXE(2)P were submitted to jugular cannulation and LC electrolytic lesion or sham-operation, at 09:00 h. Blood samples were collected hourly from 11:00 to 18:00 h, when animals were decapitated and their brains removed to analyze LC lesion and punch out the MPOA and ME. Plasma LH levels and LHRH content of MPOA and ME were determined by radioimmunoassay. During metestrus, diestrus and estrus, LC lesion did not modify either LH plasma concentrations or LHRH content, but completely abolished the preovulatory LH surge during proestrus and the surge of OVXE(2)P. These blockades were accompanied by an increased content of LHRH in the MPOA and ME. The results suggest that: (1). LC does not participate in the control of basal LH secretion but its activation is essential to trigger spontaneous or induced LH surges, and (2). the increased content of LHRH in the MPOA and ME may be due to a decreased NE input to these areas. Thus, LC activation may be required for depolarization of LHRH neurons and consequent LH surges.     

Preoptic injection of VIP, but not of secretin or PHI, inhibits LH and stimulates prolactin secretion in the ovariectomized rat

The microinjection of vasoactive intestinal peptide (VIP) into the medial preoptic area rapidly inhibited pulsatile luteinizing hormone (LH) secretion within 1 h in ovariectomized rats. A transient but significant increase in prolactin (PRL) secretion occurred after VIP injection coincidentally with the initiation of the inhibition of LH pulses. Preoptic microinjections of secretin or peptide histidine isoleucine amide (PHI) did not have such effects. These results suggest specific actions of VIP, despite its structural similarity to secretin and PHI, on the preoptic mechanisms governing LH and PRL secretion in ovariectomized rats.     

Locus coeruleus lesions decrease norepinephrine input into the medial preoptic area and medial basal hypothalamus and block the LH, FSH and prolactin preovulatory surge

The aim of this work was to study the role of the dorsal noradrenergic ascending pathway (DNAP), which originates in the locus coeruleus (LC) on the preovulatory surge of luteinizing hormone (LH) follicle-stimulating hormone (FSH) and prolactin (PRL) by producing bilateral electrolytic lesions (cathodal or anodal) in this nucleus. LC lesions were placed at 11.00 h on proestrus in female rats with regular 4-day estrous cycles. Intact rats, sham-operated as well as animals with missed lesions served as controls. In Experiment I, anodal current was applied and hourly blood samples were withdrawn (from 13.00 to 17.00 h) via a jugular catheter from conscious, freely moving rats for determination of plasma LH, FSH and PRL concentrations. In Expt. II, Expt. I was repeated using cathodal current and collecting blood samples hourly from 13.00 to 18.00 h. In both experiments the animals were sacrificed on the next morning when the occurrence of ovulation was checked. The medial septal area (MSA), medial preoptic area (MPOA), and medial basal hypothalamus (MBH) were dissected and assayed for norepinephrine (NE), dopamine (DA) and 5-hydroxyindoleacetic acid (5-HIAA) content. Experiment III was performed in order to test if a hormonal discharge occurred immediately after lesion placement. Blood samples were collected immediately before and 15, 30, 60 and 90 min postoperatively (from 11.00 to 12.30 h). Either anodal or cathodal lesions blocked the proestrous surge of LH, FSH and PRL. The hypothesis that the lesions advanced or delayed these hormonal surges was rejected since we found no increases in the hormonal levels from 11.00 to 12.30 or from 13.00 to 18.00 h, and ovulation was not observed on the following morning in the lesioned animals. Since control, sham-operated and missed-lesion groups exhibited LH, FSH and PRL surges and ovulation, this blockage appears to be caused by the destruction of the LC neurons. Also, this blockade was correlated with a decrease in the NA content in the MPOA and MBH, but not in the MSA, whereas the DA and 5-HIAA content were not changed in all groups examined. The results lead us to suggest that the integrity of noradrenergic afferent input from the LC to luteinizing hormone-releasing hormone neurons in the MPOA and MBH is essential for triggering the preovulatory surge mechanisms for gonadotrophins and PRL.     

Involvement of central catecholamines in the feedback actions of 17beta-estradiolbenzoate on luteinizing hormone secretion in the ovariectomized female rat.

(1) Evidence has been presented, based on quantitative microfluorimetric estimations of dopamine (DA) and noradrenaline (NA) levels and turnover, and on radioimmunological measurements of serum luteinizing hormone (LH), and follicle-stimulating hormone (FSH) and prolactin levels, that the central inhibitory feedback action of estradiol on LH secretion mainly involves a marked increase in DA turnover of the lateral palisade zone (LPZ) of the median eminence and also involves a reduction of NA turnover, mainly located in the medial preoptic area (MPOA) and in the subependymal layer (SEL). (2) The mechanism for these changes in catecholamine (CA) turnover is discussed. The possibility is favored that they involve the stimulation of central cytosol estrogen receptors which may even be located in the arcuate DA and possibly reticular NA cell bodies themselves. (3) The marked and long-lasting hypersecretion of prolactin caused by estrogen could be involved in mediating the sustained increase of DA turnover in the median eminence and the sustained reduction of NA turnover. (4) The central facilitatory feedback action of estrogen, on the other hand, may be mainly responsible for the sharp increase of NA turnover in the MPOA and SEL and also the associated reduction of DA turnover in the LPZ in the critical period of both adult cyclic rats and of immature female rats treated with PMS. (5) We take the view that these latter NA and DA turnover changes take precedence over cytosol estrogen receptors, which previous evidence indicates (see Sawyer, 1975) are located in preoptic and amygdaloid regions. (These estradiol concentrating neurons then directly and/or indirectly make connections with the NA and DA pathways). (6) FSH secretion is not controlled by DA and NA pathways. (7) The hypothesis given above is based on the assumption that different estradiol concentrating neurons are involved in the central inhibitory and facilitatory feedback action of estradiol and that the estrogen receptors of these respective neurons have differences which allow their differential activation, the inhibitory feedback leading mainly to a marked increase in the ratio DA activity/NA activity in the median eminence, whereas the facilitatory feedback causes a marked reduction of this ratio.     

The role of hypothalamic serotonin (5HT) before ovulation in immature rats treated with pregnant mare serum (PMS).

(1) PCPA methyl ester (10 mg/rat i.p.) inhibits induced ovulation in immature rats treated with pregnant mare serum (PMS). It also suppresses the preovulatory surges of LH and FSH, but not those of oestradiol or progesterone. (2) There is an increase in hypothalamic 5HT levels in the aftermoon and hypothalamic 5HIAA levels in the evening of the two days studied (days 28 and 29 of life). This occurs whether or not PMS was given on day 27. (3) The antiovulatory effects of PCPA are only seen when it is given on the afternoon or evening of the day before the pre-ovulatory gonadotrophin surge, i.e. on day 28 over the period of raised hypothalamic 5HT metabolism. (4) PCPA reduces 5HT metabolism in the hypothalamus within 2 hr of administration and its anti-ovulatory effect can be overcome by 5-hydroxytryptophan. This indicates that hypothalamic 5HT activity is essential for the gonadotrophin surge. (5) The anti-ovulatory effect of PCPA can be overcome by progesterone, LH and FSH but not oestradiol.     

Prolactin secretory surge during estrus coincides with increased dopamine activity in the hypothalamus and preoptic area and is not altered by ovariectomy on proestrus

Prolactin (PRL) secretory surges have been reported on the afternoons of both proestrus and estrous in cycling rats. As neuroendocrine regulation of estrous PRL surge is poorly understood, the present study aimed to investigate the involvement of hypothalamic dopamine and serotonin as well as of plasma ovarian steroids in this hormonal surge generation. For that, we determined the concentrations of dopamine, serotonin and their respective metabolites 3,4-dihydroxyphenylacetic acid (DOPAC) and 5-hydroxyindole-3-acetic acid (5-HIAA) in the mediobasal hypothalamus (MBH) and medial preoptic area (MPOA) throughout the day of estrus and correlated them with plasma PRL levels. In a second study, we evaluated the effect of ovariectomy on the morning of proestrus on PRL surges of both proestrus and estrus. Dopamine turnover, as determined by DOPAC/dopamine ratio, increased in both the MBH and MPOA coinciding with the afternoon PRL surge on estrus. In contrast, both the concentration and turnover (5-HIAA/serotonin) of serotonin within these areas were unaltered during estrus. In addition, ovariectomy reduced plasma estradiol and progesterone levels but did not alter the PRL surges on proestrus and estrus. Considering that dopamine is the main inhibitor of PRL release and that PRL auto-regulates its secretion through a short-loop feedback mechanism, our present results suggest that PRL may suppress its own secretion during the estrus surge through the activation of the dopaminergic neurons in the MBH and MPOA. In addition, the PRL surge on estrus seems do not depend on either the activity of hypothalamic serotonin or the increased secretion of ovarian steroids on proestrus.     

Monoamine Levels and Turnover in Brain: Relationship to Priming Actions of Estrogen

Norepinephrine (NE), dopamine (DA) and serotonin (5HT) levels and turnover rates were studied in 8 discrete brain nuclei of ovariectomized rats 24 hours after the administration of 5 microgram of estradiol benzoate (EB) or sesame oil vehicle. This estrogen paradigm, by itself, did not induce sexual behavior or alter LH levels at the time these parameters were evaluated. However, combined with progesterone, the estrogen treatment was sufficient to generate an LH surge and induce sexual receptivity. Steady state concentrations of NE were significantly higher in the diagonal band of Broca (NDB) and the periventricular nucleus (PVE2; anterior hypothalamic level) following EB treatment. In addition, 5-hydroxyindole acetic acid (5HIAA) concentrations were elevated in the dorsal raphe of EB treated animals. Estrogen did not affect steady state concentrations of DA or 5HT in any of the brain nuclei studied. Turnover rates (K, pg/microgram protein/hr) and rate constants (k, hr-1) for NE were increased in the lateral septum (K, 140%; k, 120%), NDB (K, 160%; k, 130%) and the PVE2 (K, 140%; k, 70%) in EB treated animals. Estrogen decreased the rate constant for NE by 30% in the medial preoptic area. In contrast, DA and 5HT turnover rates were not significantly affected by estrogen. These results localize sites where estrogen induces changes in noradrenergic activity and suggest that these changes may be involved in the priming action of the steroid in inducing sexual behavior and/or gonadotropin secretion.     

Estrous cycle modulation of extracellular serotonin in mediobasal hypothalamus: role of the serotonin transporter and terminal autoreceptors.

In vivo microdialysis was used to examine extracellular serotonin (5-HT) in the mediobasal hypothalamus (MBH) of male and female Fischer (CDF-344) rats. Females from the stages of diestrus, proestrus, and estrus were used. Additionally, ovariectomized rats, primed subcutaneously (s.c.) with estradiol benzoate or estradiol benzoate plus progesterone were examined. Extracellular 5-HT in the MBH varied with stage of the estrous cycle and with the light/dark cycle. Proestrous females had the highest microdialysate concentrations of 5-HT during the light portion of the light/dark cycle and lowest concentrations during the dark portion of the cycle. Diestrous females had the highest levels during the dark portion of the cycle, while males and estrous females showed little change between light and dark portions of the cycle. In ovariectomized rats, there was no effect of 2.5 microg or 25 microg estradiol benzoate (s.c.) on extracellular 5-HT; but the addition of 500 microg progesterone, 48 h after estrogen priming, reduced microdialysate 5-HT near the threshold for detection. In intact females and in males, reverse perfusion with 3 microM fluoxetine, a selective serotonin reuptake inhibitor (SSRI), or 2 microM methiothepin, a 5-HT receptor antagonist, increased microdialysate concentrations of 5-HT. Estrous females and males showed nearly a 4-fold increase in microdialysate 5-HT in response to fluoxetine while smaller responses were seen in diestrous and proestrous rats. In contrast, proestrous rats showed the largest response to methiothepin. Estrous females showed a delayed response to methiothepin, but there was no methiothepin-induced increase in extracellular 5-HT in males. These findings are discussed in reference to the suggestion that extracellular 5-HT in the MBH is regulated in a manner that is gender and estrous cycle dependent. The 5-HT terminal autoreceptor may exert a greater role in proestrous females; the serotonin transporter appears to play a more active role in the regulation of extracellular 5-HT in estrous females and in males.     

Ovarian steroids influence the activity of neuroendocrine dopaminergic neurons

The secretion of prolactin (PRL) from the anterior lobe (AL) of the pituitary gland is tonically inhibited by dopamine (DA) of hypothalamic origin. While ovarian steroids play a role in the regulation of the secretion of PRL, their effect on all three populations of hypothalamic neuroendocrine dopaminergic neurons is not fully understood. In this study we describe the effects of ovarian steroids on regulation of the release of DA from tuberoinfundibular dopaminergic (TIDA), tuberohypophyseal dopaminergic (THDA) and periventricular-hypophyseal dopaminergic (PHDA) neurons. Adult female rats were bilaterally ovariectomized (OVX) and, 10 days following ovariectomy (day 0), injected with corn oil (vehicle), estrogen, or estrogen plus progesterone (day 1). Animals were sacrificed every 2 h from 09.00 to 21.00 h by rapid decapitation. Trunk blood was collected and the concentration of PRL in serum was determined by radioimmunoassay. The median eminence (ME) and the AL, intermediate (IL) and neural (NL) lobes of the pituitary gland were dissected and the concentration of DA and DOPAC in each was measured by HPLC-EC. OVX rats presented small but significant increases in the secretion of PRL at 15.00 and 17.00 h. Replacement of estrogen or estrogen plus progesterone increased the basal concentration of PRL. Moreover, injection of estrogen only, or estrogen plus progesterone increased the concentration of PRL in serum at 15.00 h through 19.00 h, respectively, followed by a decrease to baseline thereafter. The turnover of DA in the ME and NL of OVX rats increased at 13.00 and returned to low levels. Turnover of DA in the IL of OVX rats increased in the morning by 11.00 h and remained elevated before decreasing by 17.00 h. The turnover of DA in the ME, IL and NL of OVX rats increased by 19.00 h. Injection of estrogen advanced the increase of TIDA activity by 2 h in the ME compared to OVX rats. Moreover, administration of estrogen suppressed the activity of THDA and PHDA neurons in the afternoon compared to OVX rats. In estrogen plus progesterone-treated rats, the activity of hypothalamic neuroendocrine dopaminergic neurons terminating in the ME, IL, and NL was inhibited prior to the increase in the secretion of PRL. The concentration of DA in the AL diminished prior to the estrogen-induced increase of PRL. Administration of progesterone, in concert with estrogen, delayed the increase of PRL in serum and the decrease of DA in the AL, compared to estrogen-treated rats, by 4 h. These data suggest a major role for ovarian steroids in controlling increases in the secretion of PRL by not only stimulating PRL release from lactotrophs, but also by inhibiting the activity of all three populations of hypothalamic neuroendocrine DAergic neurons.     

Lack of effect of hyperprolactinemia on serotonin turnover in ovariectomized and ovariectomized estrogen-treated rats

Hyperprolactinemia suppresses luteinizing hormone (LH) and prolactin (PRL) secretion under a variety of experimental conditions. The secretion of both of these hormones is regulated at the hypothalamic level by several neurotransmitters, including serotonin (5-HT). Therefore, we examined the effect of hyperprolactinemia on 5-HT neuronal activity in key hypothalamic areas that are rich in 5-HT terminals and which are known to regulate the release of LH and PRL. Young cycling virgin rats were ovariectomized (day 0). From days 11-16, animals were injected with ovine prolactin (oPRL, 4 mg/kg, s.c.) or vehicle every 8 h. On day 14, one-half of the oPRL- and vehicle-treated rats were implanted with 20-mm long Silastic capsules containing estradiol (180 micrograms/ml). On day 16, animals were killed at 08.00, 12.00 or 18.00 h or treated with pargyline (75 mg/kg) and killed 10 min later. Trunk blood was collected and serum was radioimmunoassayed for LH and endogenous rat PRL (rPRL). Brains were removed, frozen, sectioned and the medial preoptic, suprachiasmatic, and arcuate nuclei, median eminence and globus pallidus were microdissected. Serotonin was measured using high pressure liquid chromatographic methodology. We were unable to detect any feedback effect of hyperprolactinemia on 5-HT turnover in any brain area of ovariectomized or ovariectomized estradiol-treated rats at any time of day that we examined. Several potential reasons for the absence of an effect of hyperprolactinemia on serotonergic function are discussed.     

The participation of histaminergic receptors of the rostral hypothalamus on the tonic release of luteinizing hormone (LH) in adult spayed rats under estrogen and progesterone treatment

Summary The influence of histaminergic sites in the preoptic-anterior hypothalamic area (POA-AHA) on the basal release of luteinizing hormone (LH) under a continuous regimen of estradiol, progesterone, or both was studied in ovariectomized rats. Different groups of animals were subjected to the following experimental schedule: at day 1, rats received a s.c. silastic implant filled with oil, estradiol, progesterone, or estradiol plus progesterone. Seven days later (day 7), animals were implanted into the POA-AHA with microinjection cannulae. At day 8 and 9, the different groups of rats were microinjected with 1 l of saline solution containing 35 nMol of pyrilamine or metiamide, or 20 nMol of alpha-fluoro-methyl-histidine. At day 10, blood samples were taken through a permanent jugular cannulae implanted in situ the day before. LH concentrations were determined in plasma by RIA. Results showed that the increase of LH plasma levels induced by the ovariectomy was inhibited by the estrogen implant, as expected. Treatment of metiamide or alpha-fluoro-methyl-histidine did not affect the pattern of LH secretion. Nevertheless, treatment of metiamide induced a transient increase in the gonadotropin concentrations that extended for two hours (16:00 and 17:00 H). No change in LH plasma levels was observed in rats bearing the progesterone implant. Treatments (pyrilamine, metiamide, or alpha-fluoro-methyl-histidine into the POA-AHA) had no effect. The transient increase in the hormone levels observed in rats treated with pyrilamine in the estrogen-implanted rats was absent in rats bearing the estrogen-progesterone implant. Present data support the concept that histamine is involved in the POA-AHA to control the pituitary LH release and emphasize the role of plasma estrogen to facilitate the expression of HA receptors.     

Effect of estrogen on the lordosis-inhibiting action of ketanserin and SB 206553

The effects of the 5-HT(2A/2C) receptor antagonist, ketanserin, and the 5-HT(2C) receptor antagonist, SB 206553, on lordosis behavior were investigated in ovariectomized rats hormonally primed with estradiol benzoate (EB) (0.5 or 25 microg) and progesterone (500 microg). Both ketanserin and SB 206553 inhibited lordosis behavior after infusion into the ventromedial nucleus of the hypothalamus (VMN), but ketanserin was slightly more effective than the 5-HT(2C) receptor antagonist. Either drug was more effective in rats primed with 0.5 microg EB than in rats hormonally primed with 25 microg EB. These findings support the suggestion that estrogen may enhance functioning of the 5-HT(2) receptor family and thereby protect against the 5-HT(2) receptor antagonists. These data are consistent with prior suggestions that estrogen modulates functioning of 5-HT(2) receptors within the VMN and that 5-HT(2) receptors play a facilitatory role in the modulation of female rat lordosis behavior.     

Acute Action of Oestrogen on Medial Preoptic Gamma-Aminobutyric Acid Neurons: Correlation with Oestrogen Negative Feedback on Luteinizing Hormone Secretion

The acute effects of oestrogen on the medial preoptic area (MPOA) γ-aminobutyric acid (GABA) system were examined by delivering an intravenous bolus of 17β-oestradiol (5 μg/100 g body wt) to conscious ovariectomized rats implanted with microdialysis probes. Fifteen-min blood samples were taken to determine the time-course of negative feedback effects of oestrogen on luteinizing hormone (LH) secretion. Two h after administration of 17β-oestradiol, GABA release from the MPOA was significantly elevated compared with vehicle-treated controls (P<0.05). The rise in GABA levels continued until the end of the experiment, 4 h after 17β- oestradiol, at which time it was over 50% higher than controls (P<0.01). The pulsatile pattern of LH secretion was significantly depressed 2 and 3 h after administration of 17β-oestradiol compared with controls (P<0.05). To determine the effects of the 17β-oestradiol treatment on pituitary responsiveness to LH-releasing hormone (LHRH), a further group of rats were given exogenous LHRH (50ng/100g body wt, intravenously) before and 3 h after vehicle or 17β-oestradiol treatment and blood samples taken to determine the effect on LH secretion. The maximal LH response to LHRH in 17β-oestradiol-treated rats was approximately 50% that of control-treated values. This study demonstrates the acute and potent action of 17β-oestradiol on GABA release in the MPOA and lends support to a genomic site of action for oestrogen in modulating neural elements regulating GABA release from the MPOA. These results, showing a parallel decrease in LH secretion with increased GABA levels in the MPOA, suggest a role for GABA elements within the MPOA as a site of oestrogen negative feedback on LH secretion.     

A Comparison of the Effects of Medial Preoptic Electrical Versus Electrochemical Stimulation on Luteinizing Hormone-Releasing Hormone Neuronal Responsiveness to Norepinephrine

Recently, we reported that luteinizing hormone-releasing hormone (LHRH) neurons of estrogen-treated, ovariectomized rats have only limited responsiveness to norepinephrine (NE). These conclusions were based upon observations that NE, when infused intracerebroventricularly, produced only minor increases in plasma luteinizing hormone (LH), whereas, similar infusions following preliminary medial preoptic area (MPOA) electrochemical stimulation (ECS) markedly amplified LH secretion. One difficulty with this approach is that ECS produces an irritative lesion and deposits iron within the tissue, whereas, electrical stimulation (ES) does not have such effects. Accordingly, in the present study, we compared the effects of MPOA–ECS versus –ES on LHRH neuronal responsiveness to NE. While equivalent peak LH concentrations occurred within 15 min after MPOA–ECS or –ES, in the ECS group, LH release was sustained, whereas, it abruptly ceased upon termination of ES (at 15 min). The intracerebroventricular pulse infusion of NE at the time of peak LH secretion (30 min) in MPOA–ECS animals markedly amplified LH release. In these animals, plasma LH remained significantly elevated for 75 min before a decline was observed. In contrast, an infusion of NE at the time of maximal LH release in ES rats (16 min) did not augment LH secretion. The second series of studies examined the effects of MPOA infusions of NE in animals receiving preoptic ES. A single infusion of NE 16 min after ES (i.e. one min after termination of ES) did not amplify LH release, but when two NE pulses were given at 5 and 16 min after beginning preoptic ES, peak plasma LH levels were maintained for an additional 30 min before a decline occurred. Pretreatment of rats with a yS-adrenoreceptor antagonist (propranolol) or a monoamine oxidase inhibitor did not affect peak LH responses obtained after either MPOA–ES alone or combined with two pulses of NE infused into the MPOA at 5 and 16 min. We conclude that following cessation of MPOA–ES, LHRH neurons rapidly lose their responsiveness to NE, whereas, rats which received MPOA–ECS retain such responsiveness possibly due to the stimulative properties of the iron deposited by the ECS. Presumably, for NE to trigger an LH surge requires prior removal of some intrinsic inhibitory control which regulates LHRH neuronal responsiveness to NE.     

Steroid-induced alterations in mRNA expression of the long form of the prolactin receptor in the medial preoptic area of female rats: Effects of exposure to a pregnancy-like regimen of progesterone and estradiol

It is firmly established that the onset of maternal behavior in the female rat is stimulated by a combination of hormones that include prolactin (PRL), estradiol (E2), and progesterone (P(4)). Specifically, nulliparous rats display short latencies to respond to foster young when primed with Silastic capsules filled with P4 and E2 and then administered PRL centrally to the medial preoptic area (MPOA), an area integrally involved in the expression of maternal behavior in this species. PRL or P4 treatments alone are ineffective in stimulating the expression of maternal care. Since the actions of PRL in the MPOA appear to be mediated by PRL receptors, it was of interest to determine whether and how treatment with P4 and E2 together or separately might alter mRNA expression of the long form of the PRL receptor (PRL-R(L)) in the MPOA. Using in situ hybridization histochemistry (ISHH), mRNA expression of the PRL-R(L) was measured in the MPOA of ovariectomized, nulliparous rats treated with various combinations of P4 and E2. Treatment of animals with P4 alone for 10 days or with P4 followed by E2 for 1 or 4 days resulted in reductions in PRL receptor mRNA expression in the MPOA when compared with the expression in animals treated with E2 alone or blank capsules. The actions of P4 on mRNA expression of the PRL-R(L) were more pronounced in the dorsal MPOA. Circulating PRL levels collected at the time of sacrifice were elevated in all groups treated with E2, but no association between PRL levels and receptor mRNA expression within the MPOA was evident. These findings indicate that the dorsal MPOA may be one site of progesterone's action in facilitating prolactin-mediated maternal behavior.     

Effect of Hypothalamic Deafferentation on the Pulsatile Secretion of Luteinizing Hormone in Ovariectomized Lactating Rats*

The pulsatile luteinizing hormone (LH) secretion in ovariectomized lactating rats bearing complete (CD), anterior (AD), anterolateral (ALD), posterior (PD), or roof (RD) deafferentation of the hypothalamus was determined. All lactating rats were ovariectomized on Day 2 of lactation (Day 0, day of parturition). The deafferentation of nerve fibres to the mediobasal hypothalamus was performed on Day 6 or 7 of lactation. Twenty-four h after the surgery, blood samples were taken through the indwelling atrial catheter every 6 min for 3 h. Plasma concentrations of LH and prolactin (PRL) were measured by radioimmunoassay. The loss of LH pulses associated with lactation was still apparent following AD, PD and sham-deafferentation (SD); pulsatile LH secretion was, however, present in rats with CD, ALD and RD despite continued suckling. The only significant difference in plasma PRL concentrations among the various groups was a reduction in the PRL level in rats with RD in comparison to those with SD. We conclude that the neural signal responsible for the inhibition of pulsatile LH release by suckling is conveyed through the dorsal part of the hypothalamus and PRL does not mediate the suppression of LH pulses in mid-lactation.     

Role of serotonin in estrogen-progesterone induced luteinizing hormone release in ovariectomized rats

Pharmacological agents were used to manipulate the surge of luteinizing hormone (LH) induced by progesterone in ovariectomized rats primed with estradiol benzoate. The LH surge was abolished with p-chlorophenylalanine (PCPA), an inhibitor of tryptophan hydroxylase, and restored by 5-hydroxytryptophan, a serotonin precursor. Serotonin receptor agonists, quipazine and N-N-dimethyl-5-methoxytryptamine, were also capable of inducing an LH surge in rats pretreated with PCPA. The serotonin reuptake blocker chlorimipramine was ineffective in stimulating LH release in PCPA blocked animals. Another reuptake blocker, zimelidine was only partially effective in this regard. These two reuptake blockers, as well as amitriptyline, when injected to non-PCPA treated rats led to the reduction or inhibition of the expected LH surge. Four serotonin receptor antagonists, cyproheptadine, methysergide, cinanserin and SQ-10,631, were each able to reduce or abolish the progesterone induced surge of LH. These results suggest that some of the reuptake blockers of serotonin are also capable of inhibiting receptor binding for this neurotransmitter and strongly indicate that serotonin has a stimulatory role in the steroid induced release of LH in castrated rats.     

LHRH release depends on Locus Coeruleus noradrenergic inputs to the medial preoptic area and median eminence

We tested the hypothesis that Locus Coeruleus (LC) inputs to the medial preoptic area (MPOA) and median eminence (ME) are essential for gonadotropin release. Proestrus and ovariectomized (OVX) rats were decapitated at 16:00 h. LC electrolytic lesion was performed at 11:00 h during proestrus and 24h before decapitation in OVX rats. Plasma luteinizing hormone (LH) and follicle stimulating hormone (FSH) were measured and MPOA and ME were microdissected for LHRH content measurement. In addition, FOS protein in LC and MPOA were studied in proestrus and OVX rats at 12:00, 15:00, and 17:00 h. On proestrus, LC lesion blocked the LH surge and only decreased plasma FSH; in OVX rats the lesion induced only a slight decrease on plasma LH without affecting FSH secretion. An increased content of LHRH in the MPOA and ME of both groups accompanied the decreases of plasma LH. In proestrus, the number of FOS-immunoreactive (FOS-ir) neurons increased from 12:00 to 17:00 h in the LC and MPOA. In OVX rats, there was an increase at 15:00 h in the LC and a decrease at 17:00 h in both areas. The number of FOS-ir neurons was lower in OVX than in proestrus animals. Thus, LC (1) is responsible, at least in part, for gonadotropin release through the activation of LHRH neurons, (2) is more closely related to the positive than the negative feedback, and (3) seems to show an intrinsic cyclic activity which is amplified by ovarian steroids.