Hamster refractoriness: the role of insensitivity of pineal target tissues.

Hamsters exposed to short days undergo gonadal collapse followed by recrudescence and insensitivity to the regressive effects of such photoperiods. This refractoriness may be due to exhaustion of the pineal gland or desensitization of its target. Hamsters whose gonads had spontaneously recrudesced were injected with melatonin (25 micrograms per injection) once daily (known to induce regression in intact hamsters) or twice daily (reported to arrest reproduction in pinealectomized hamsters) for 7 weeks. In neither case did refractory hamsters respond to melatonin treatment. The gonads of intact hamsters treated with melatonin for 21 weeks regressed and spontaneously recrudesced along a normal time course. These results indicate that gonadal refractoriness is due to insensitivity of the target tissues of the pineal gland and imply that melantonin participates in photoperiodic regulation of reproduction in the golden hamster.

Regulation of the phase and period of circadian rhythms restored by suprachiasmatic transplants.

The influence of exogenous signals on circadian rhythms restored by transplants of the suprachiasmatic nucleus (SCN) of the hypothalamus has received little study. The authors tested the responsiveness of hamsters bearing SCN transplants to photic and pharmacological treatments. Light intensities as high as 6,500 lux were insufficient to produce entrainment, although masking was observed frequently. Triazolam failed to produce statistically significant phase shifts when administered during the subjective day, but 2 animals bearing functional SCN grafts responded to this benzodiazapine during the subjective night. The authors next tested the hypothesis that the host can retain circadian aftereffects that influence the period of the circadian system reconstituted by the graft. Intact hamsters were entrained to light:dark cycles of short (23.25-h) and long (25-h) period (T) for at least 3 months. Control hamsters released into constant darkness exhibited profound and long-lasting aftereffects of entrainment to T cycles. Hamsters that received SCN lesions after exposure to these T cycles and SCN grafts 3 weeks later exhibited marginal but statistically significant aftereffects that disappeared within 3 months. On subsequent transfer to constant light, tau lengthened by 0.25 +/- 0.6 h in hamsters with intact SCN (p < .05). Animals bearing SCN grafts continued to free run in constant light but differed from intact animals in that circadian period did not lengthen. Functional SCN grafts contained vasoactive intestinal polypeptide (VIP), neurophysin (NP), and cholecystokinin (CCK) immunoreactive (ir) cells. Inputs of neuropeptide Y-and serotonin-ir fibers from the host brain to grafted SCN peptide cell clusters were variable. Limited observations using retrograde and anterograde tracers do not support the existence of extensive input to the graft. Retinal input overlapped only rarely with clusters of VIP-ir, CCK-ir, or NP-ir cells. The authors conclude that the circadian system reinstated by SCN transplants is relatively impervious to photic influences that exert parametric and nonparametric influences in intact hamsters. The transient expression of aftereffects induced in the host before transplantation indicates that extra-SCN systems of the host can influence the period of the reconstituted circadian system to at least a limited degree.

Pineal melatonin secretion drives the reproductive response to daylength in the ewe.

This study was conducted to determine whether the pineal indoleamine melatonin mediates the effects of photoperiod on the capacity of estradiol to inhibit LH secretion in the ewe. Patterns of serum melatonin were characterized in pineal-intact ovariectomized ewes treated with sc Silastic estradiol implants and exposed to 90-day alternations between long and short photoperiods. High fluctuating levels of serum melatonin were found during the night, with the duration of elevated serum levels corresponding to the length of the dark period. Transfer from long to short photoperiods caused a rapid change in the duration of nightly melatonin secretion and reduced the negative feedback potency of estradiol upon LH secretion during the natural anestrous season. In pinealectomized ewes, the night-time rise of melatonin was absent, and transfer from long to short days failed to reverse the capacity of estradiol to inhibit LH secretion during anestrus. Nightly infusions of melatonin restored patterns of this indoleamine similar to those observed in pineal-intact ewes exposed to the 90-day alternation between long and short days. The melatonin infusions also restored the reproductive response to the inductive photoperiod: in every ewe, the negative feedback effects of estradiol upon LH secretion were diminished after transfer from long to short days. The amplitude and latency of this escape matched those of pineal-intact animals. We conclude that the pineal mediates the reproductive response of the ewe to inductive photoperiods through its daily rhythm of melatonin secretion.

Role of the pineal gland in ovine photoperiodism: regulation of seasonal breeding and negative feedback effects of estradiol upon luteinizing hormone secretion.

In this study we determined whether the pineal is required for the photoperiodic control of reproduction in ewes and, if so, whether its effect is mediated via negative feedback effects of estradiol on LH. Two groups of Suffolk ewes were pinealectomized and challenged with 90-day alternations between long (16 h of light, 8 h of darkness) and short (8 h of light, 16 h of darkness) photoperiods for 2 yr. One group of pinealectomized ewes had intact ovaries; the other was ovariectomized and implanted with estradiol to monitor feedback inhibition of LH. Reproductive effects of pinealectomy were correlated with elimination of the nighttime rise of serum melatonin. In pineal-intact control ewes, reproductive function was readily manipulated by photoperiod. Long days inhibited ovarian cyclicity and increased the negative feedback potency of estradiol regardless of time of year; short days had stimulatory effects on both variables. In most animals, pinealectomy abolished both the inductive effects of short days and the inhibitory effects of long days, and eliminated the nighttime rise in serum melatonin. Although seasonal changes in reproductive function persisted after pinealectomy, the timing of these changes coincided most closely with that of pineal-intact animals housed outdoors. With the passage of time after pinealectomy, however, these seasonal fluctuations became progressively damped. These results document pineal mediation of the photoperiodic control of seasonal breeding in ewes and indicate that the pineal exerts its effect, at least in part, by changing the capacity of estradiol to inhibit LH secretion. Furthermore, the pineal mediates the response to both stimulatory and inhibitory photoperiods. Finally, ewes that are unresponsive to photoperiod remain seasonal, either in response to other environmental cues or owing to the expression of an endogenous circannual rhythm of reproduction.

Effects of photoperiod and androgen on proopiomelanocortin gene expression in the arcuate nucleus of golden hamsters.

In golden hamsters, seasonal changes in day length act via a pineal-dependent mechanism to regulate feedback and behavioral effects of androgen. Endogenous opiates participate in photoperiodically regulated neuroendocrine functions, but the effects of androgen on expression of the gene encoding POMC, the precursor of beta-endorphin, have been controversial. We used quantitative in situ hybridization to examine regulation of POMC messenger RNA (mRNA) by testosterone and to test the hypothesis that short day lengths act through the pineal gland to amplify POMC mRNA expression. We studied intact hamsters and castrates with or without androgen treatment held in long (14 h of light, 10 h of darkness) or short (5 h of light, 19 h of darkness) days for 10 weeks. POMC gene expression differed with rostral-caudal plane, photoperiod, and surgical treatment (castration and testosterone administration). Testosterone increased the number of silver grains in labeled cells throughout the arcuate nucleus, and short day castrates given androgen consistently had more silver grains per labeled cell than did their long day counterparts. Testosterone exerted an inhibitory effect, however, on the number of POMC mRNA-positive cells, and more POMC mRNA-labeled cells were found in the arcuate nucleus of long than short day castrates treated with testosterone. Photoperiod had no significant influence in castrates not receiving androgen. Testosterone treatment had generally similar effects whether it was begun at the time of castration or 5 weeks later. Pinealectomy blocked the influence of photoperiod on both the mean number of silver grains per labeled cell and the number of labeled cells. The results indicate that day length regulates POMC gene expression when androgen levels are held constant, but that androgen is necessary for photoperiod effects to be expressed.

A role for estradiol in enhancing luteinizing hormone pulse frequency during the follicular phase of the estrous cycle of sheep.

Experiments were conducted to test the hypothesis that the increased frequency of LH pulses during the follicular phase of the sheep estrous cycle can be explained by the withdrawal of progesterone. This steroid imposes a profound inhibition of LH pulse frequency in the luteal phase. Experimental ewes were ovariectomized in the late luteal phase of the estrous cycle and divided into three groups: 1) no estradiol provided; 2) basal estradiol maintained at 1-2 pg/ml by small sc Silastic estradiol implants; and 3) peak estradiol of 5-6 pg/ml provided by larger estradiol implants. Control ewes had intact ovaries; their follicular phases were synchronized by insertion and subsequent removal of progesterone implants. LH pulses were monitored beginning 24 h after ovariectomy of experimental ewes or progesterone implant removal from intact control ewes. In the follicular phase controls, LH pulse frequency increased 3- to 4-fold after progesterone withdrawal, reaching up to two pulses per h. When estradiol was not provided after ovariectomy of experimental ewes, LH pulse frequency also increased, but not to the extent seen in the follicular phase control. This high frequency was achieved, however, in experimental ewes treated with either basal or peak estradiol. Both estradiol treatments also reduced LH pulse amplitude. These results fail to support the hypothesis that the high frequency of LH pulses in the follicular phase is solely a consequence of progesterone withdrawal at luteolysis. Rather, they suggest that estradiol (but not necessarily rising estradiol) also contributes to the high frequency pulses of LH that occur in the ewe at this time.

The endocrine basis of the synergistic suppression of luteinizing hormone by estradiol and progesterone.

The basis of the synergism between estradiol and progesterone in suppressing tonic (pulsatile) LH secretion was examined in the ewe, making use of the observation that progesterone exerts its inhibition selectively on LH pulse frequency, while estradiol decreases only pulse amplitude. To accomplish this, we analyzed changes in LH pulse patterns produced by a low level of progesterone in ovariectomized ewes treated with Silastic estradiol implants from the time of gonadectomy. A serum progesterone level of about 1 ng/ml was chosen because it inhibits LH only in the presence of estradiol. Under these circumstances, a decrease in LH pulse amplitude during progesterone treatment would suggest that progesterone increased the response to the existing level of estradiol; a decrease in pulse frequency would suggest that estradiol increases the effectiveness of progesterone. It was found that the low level of progesterone produced a decrease in LH pulse frequency in estradiol-treated ovariectomized ewes without altering pulse amplitude. These results are consistent with the hypothesis that the synergism of these two steroids reflects, at least in part, an estradiol-induced increase in the sensitivity of the central nervous system to the negative feedback action of progesterone.

Effects of suprachiasmatic transplants on circadian rhythms of neuroendocrine function in golden hamsters.

Grafts of fetal tissue including the suprachiasmatic nucleus (SCN) of the hypothalamus restore locomotor rhythmicity to behaviorally arrhythmic, SCN-lesioned Syrian hamsters. We sought to determine whether such transplants also reinstate endocrine rhythms in SCN-lesioned hamsters. In Exp 1, SCN lesions interrupted estrous cycles in a 14 h light, 10 h dark photoperiod and locomotor rhythms in constant dim red light (DD). SCN grafts that reinstated behavioral circadian rhythms consistently failed to reestablish estrous cycles. After ovariectomy, estradiol implants triggered LH surges at approximately circadian time 8 in 10 of 12 brain-intact control females and 0 of 9 SCN-lesioned, grafted females. Daily rhythms of the principal urinary melatonin metabolite, 6alpha-sulfatoxymelatonin, were not reestablished by behaviorally functional grafts. In Exp 2, SCN lesions eliminated locomotor rhythmicity in adult male hamsters maintained in DD. Seven to 12 weeks after restoration of locomotor activity rhythms by fetal grafts, hosts and sham-lesioned controls were decapitated at circadian times 4, 8, 12, 16, 20, or 24. Clear circadian rhythms of both serum corticosterone and cortisol were seen in sham-lesioned males, with peaks in late subjective day. No circadian rhythms in either adrenal hormone were evident in serum from lesioned-grafted males. Testicular regression, observed in intact and sham-lesioned males maintained in DD, was absent not only in arrhythmic SCN-lesioned hamsters given grafts of cerebral cortex, but also in animals in which hypothalamic grafts had reinstated locomotor rhythmicity. The pineal melatonin concentration rose sharply during the late subjective night in control hamsters, but not in SCN-lesioned animals bearing behaviorally effective transplants. Even though circadian rhythms of locomotor activity are restored by SCN transplants, circadian endocrine rhythms are not reestablished. Endocrine rhythms may require qualitatively different or more extensive SCN outputs than those established by fetal grafts.

Influences of photoperiod on sexual behaviour, neuroendocrine steroid receptors and adenohypophysial hormone secretion and gene expression in female golden hamsters.

Exposure to short daylengths arrests the oestrous cycle, provokes daily gonadotrophin surges and reduces the ability of exogenous oestradiol to trigger behavioural receptivity in golden hamsters. In order to examine neuroendocrine effects of photoperiod which might underlie these responses, ovariectomized hamsters were maintained under long or short photoperiods for 54 days before treatment with cholesterol or various doses of oestradiol-17 beta. Short days reduced the ability of low doses of oestrogen to prime hamsters for the induction of oestrus by progesterone. Upon repetition of oestrogen priming 2 weeks later, photoperiod was without significant influence on the concentrations of nuclear oestrogen receptors or cytosolic progestin receptors in a block of tissue containing the hypothalamus and preoptic area. Oestradiol treatment provoked significant increases in serum concentrations of LH and prolactin in the afternoon, but photoperiod did not alter the positive-feedback efficacy of this gonadal steroid hormone. Adenohypophysial LH-beta subunit and prolactin mRNAs were suppressed by short days in ovariectomized hamsters not treated with oestradiol. Oestradiol decreased expression of the LH-beta subunit gene in both stimulatory and inhibitory photoperiods, but increased prolactin mRNA abundance in both long and short days. Photoperiod therefore exerts pronounced steroid-independent effects on phasic LH and prolactin secretion, but regulation of adenohypophysial abundance of LH-beta subunit and prolactin mRNAs by oestradiol is not markedly influenced by daylength. Photoperiodic regulation of the priming effects of oestradiol on behavioural receptivity may result from modulation of events occurring subsequent to steroid-receptor interactions, or involve changes in receptor populations not detectable by the present methods.

Serum levels of gonadotrophins in hamsters exposed to short photoperiods: effects of adrenalectomy and ovariectomy.

Female hamsters rendered acyclic by exposure to short photoperiods (10 h light: 14 h darkness) showed diurnal surges of both LH and FSH which persisted after removal of the ovaries and adrenal glands. The reduced increase in gonadotrophin secretion after ovariectomy, typical of hamsters exposed to short days, was also not contingent upon the presence of the adrenal glands. The results demonstrate that 'supersensitivity' of the neuroendocrine axis to feedback of ovarian or adrenal steroids cannot fully account for either the daily surges of gonadotrophins or the diminished response of LH and FSH to ovariectomy in hamsters maintained on short days. It is suggested that these changes result instead from other pineal-mediated actions of short photoperiods upon the brain.

Regulation by the pituitary gland of circadian rhythms in the hamster.

Locomotor activity of male hamsters was recorded during long-term exposure to constant light (LL), constant darkness (DD) and during entrainment (modification of a circadian rhythm) to a 14 h light : 10 h darkness photoperiod (14L : 10D). In LL the period of the activity cycle was substantially longer in hypophysectomized than in control animals. This difference persisted during tests in DD. Although hypophysectomy reduced the duration of the active phase in some hamsters, overall the difference between the groups was not significant. The phase angle of onset of activity in 14L : 10D was not affected by hypophysectomy. Hypophysectomized female hamsters tested in DD had activity rhythms whose periods were longer than those of control animals; they were also significantly less active than corresponding controls during the first 4 h of the subjective night but the duration of the active phase did not differ significantly between the groups. These results suggest that hormones of the pituitary-gonadal axis modulate the period of circadian oscillation.

Steroidal and photoperiodic regulation of opiate binding in male golden hamsters.

This study was designed to investigate interactions between daylength and testosterone (T) in the regulation of 3H-naloxone binding which may contribute to seasonal changes in the negative feedback and behavioral effects of androgens in the golden hamster. Photoperiod influenced opiate binding in hamsters with intact gonads only in the medial amygdala. Castration elevated specific 3H-naloxone binding in the medial amygdala, medial preoptic nucleus, bed nucleus of the stria terminalis, and in the basolateral amygdaloid nucleus of hamsters exposed to either long (14L:10D) or short (5L:19D) days. Exposure to SD renders hamsters less sensitive to T maintenance in reversing these effects. Delay of T replacement until 5 weeks after castration eliminated the ability of this androgen to reverse the influence of castration upon opiate receptors in the medial amygdala. Pinealectomy markedly increased 3H-naloxone binding in short days in several brain areas. The data demonstrate that androgens and photoperiod interact to regulate 3H-naloxone binding, particularly in the medial amygdala. These effects may play a functionally relevant role in seasonal changes in the expression of sexual behavior and/or gonadotropin secretion.

Pituitary hormone gene expression in male golden hamsters: interactions between photoperiod and testosterone.

Abstract Daylength regulates neuroendocrine function in male golden hamsters. Exposure to short days triggers gonadal regression and decreases serum luteinizing hormone (LH), prolactin and testosterone concentrations. Inhibitory photoperiods also amplify the negative feedback actions of androgens upon gonadotropin secretion. To examine whether these changes arise from altered adenohypophyseal gene expression, we measured the abundance of the messenger ribonucleic acids (mRNAs) encoding beta-LH, prolactin and proopiomelanocortin in anterior pituitaries of male golden hamsters which were either left intact, castrated, castrated and implanted with testosterone, or pinealectomized and maintained in either long (14 h light/10 h dark) or short (5 h light/19 h dark) days. Short days caused testicular atrophy in intact male hamsters and reduced serum LH in intact and castrated, testosterone-replaced hamsters. The relative abundance of beta-LH mRNA was suppressed by exposure to short days only in castrated hamsters. Serum prolactin was decreased by short days regardless of circulating testosterone concentrations. Prolactin mRNA abundance was decreased by short days in all pineal-intact groups. Castration reduced proopiomelanocortin mRNA abundance in long days and testosterone replacement reversed this effect. In the presence of testosterone, photoperiod influenced serum LH concentrations without altering hypophyseal abundance of beta-LH mRNA. In contrast, photoperiodic influences on prolactin secretion were correlated with alterations in steady-state mRNA abundance.

Oestrogen receptor-alpha-immunoreactive neurones project to the suprachiasmatic nucleus of the female Syrian hamster.

Ovarian steroid hormones regulate circadian period and phase, but classical receptors for these hormones are absent in the circadian pacemaker localized in the suprachiasmatic nucleus of the hypothalamus (SCN). In order to determine whether effects of oestrogen may be exerted through steroid-binding systems afferent to the SCN we have performed double label immunocytochemistry for oestrogen receptor-alpha(ER-alpha) and the retrograde tracer cholera toxin B subunit (CtB) after its application to the SCN. Most of the areas that contain ER-alpha-immunoreactive (ERalpha-ir) cells also contained cells afferent to the SCN. The percentage of neurones afferent to the SCN which show ERalpha-immunoreactivity varies between areas. As many as one-third of the neurones afferent to the SCN in some parts of the preoptic area and the corticomedial amygdala are ERalpha-ir. Very few of the afferent neurones from the septum and the central grey are ERalpha-ir, whereas an intermediate proportion of afferents from the bed nucleus of the stria terminalis and the arcuate nucleus are ERalpha-ir. Our retrograde tracing results were compared with results of anterograde tracing from some of the sites containing SCN afferents. Using a combined retrograde and anterograde tracing technique we tested the possibility that single ERalpha-ir neurones afferent to the SCN could receive reciprocal innervation by SCN efferents. Although we found SCN input to some SCN afferent neurones, we found no evidence of reciprocity between single ERalpha-ir cells and the SCN. Our results indicate the existence of oestrogen binding systems afferent to the SCN. These neuroanatomical pathways may mediate effects of gonadal steroid hormones on circadian rhythms.

Effects of photoperiod duration and melatonin signal characteristics on the reproductive system of male Syrian hamsters.

Three experiments tested effects of photoperiod and the pineal hormone melatonin (MEL) on reproductive function among male Syrian-hamsters. In Experiment 1, hamsters were exposed for 32 weeks to 1 of 4 short photoperiods which varied in duration (11.5 L; 10 L; 8 L; 6 L). A fifth group was shifted from 11.5 L to 6 L after 6 weeks. Shorter photoperiods were associated with more rapid regression of the testes, but all groups eventually regressed to the same extent. In contrast, the temporal profile of testicular recrudescence, expressed as males became photorefractory, was not significantly different between groups. A decrease in photoperiod from 11.5 L to 6 L after 6 weeks did not delay the onset of recrudescence. The 11.5 L group was subdivided at week 32 and transferred to either 13 L or 16 L for the next 8 weeks to break photorefractoriness. Upon subsequent exposure to 8 L, both subgroups regressed their testes in similar fashion over weeks 40-52, indicating that the two long photoperiods were equally effective in breaking photorefractoriness. Nevertheless, FSH and prolactin were more consistently suppressed in the 16 L group following the switch to 8 L. Experiment 2 tested whether differing durations of MEL, administered s.c. each night for 9 weeks, elicit graded rates of reproductive regression in pinealectomized males. Testicular regression was more rapid in the group receiving MEL for 12 h than it was in the group receiving MEL for 8.5 h, thus supporting the hypothesis that the faster rates of testicular regression in the shorter photoperiods of Experiment 1 were due to their concomitant longer durations of nightly MEL secretion. Experiment 3 tested the hypothesis that rates of testicular regression in males receiving exogenous MEL would be affected by their prior photoperiodic history. Males were exposed to 18 L or 14 L for 7 weeks, then pinealectomized and administered 9.5 h MEL infusions s.c. each night for 9 weeks. In contrast to predictions, photoperiodic history had only transitory effects on MEL-induced testicular regression. Although the differences in MEL duration that accompany different short photoperiods have reproductive consequences (Experiment 1), the extent to which MEL duration expands during the transition from stimulatory to inhibitory photoperiods appears to be a less significant variable (Experiment 3).

Co-expression of melatonin (MEL1a) receptor and arginine vasopressin mRNAs in the Siberian hamster suprachiasmatic nucleus.

Durational melatonin signals, cued by the photoperiod and generated by the pineal gland, are processed in the brain to induce seasonally appropriate physiological and behavioural adaptations. The melatonin receptor subtype MEL1a (also known as mt1) appears to regulate seasonal responses. Single label in situ hybridization for MEL1a receptor mRNA revealed labelled cells in several brain regions of Siberian hamsters, including the suprachiasmatic nucleus, the paraventricular nucleus of the thalamus, and the reuniens nucleus of the thalamus. To characterize suprachiasmatic nucleus cells containing MEL1a receptor mRNA, we used 35S-labelled cRNA probes for MEL1a receptor mRNA in combination with digoxigenin-labelled cRNA probes for vasopressin, somatostatin, or orphan retinoid Z receptor beta (RZRbeta; a putative nuclear melatonin receptor). Cells in the suprachiasmatic nucleus that contained MEL1a receptor mRNA also contained mRNAs for vasopressin and RZRbeta, but not for somatostatin. These data suggest that suprachiasmatic nucleus vasopressin cells may respond to melatonin signals, raising the possibility that suprachiasmatic nucleus vasopressin output mediates some of the effects of melatonin on seasonal or circadian responses.

Regional distribution of iodomelatonin binding sites within the suprachiasmatic nucleus of the Syrian hamster and the Siberian hamster.

The pineal hormone melatonin is a potent regulator of seasonal and circadian rhythms in vertebrates. In order to characterize potential target tissues of melatonin, the distribution of iodomelatonin (IMEL)-binding sites was examined within neurochemically and anatomically defined subdivisions of the suprachiasmatic nucleus (SCN), a structure necessary for seasonal and circadian rhythms in mammals. Studies were carried out in both the adult Syrian (Mesocricetus auratus) and Siberian (Phodopus sungorus) hamster. The retinoreceptive zone of the SCN was identified anatomically by immunocytochemical (ICC) visualization of cholera toxin B subunit tracer (ChTB-ir) following its intra-ocular injection. Photically-responsive SCN cells were identified by immunostaining for the protein product of the immediate-early gene c-fos (Fos-ir) following exposure of the animal to light. The non-photoresponsive zone of the SCN was identified using in situ hybridization (ISH) for arginine vasopressin (AVP) mRNA, whilst sites of IMEL-binding in the SCN were identified by in vitro film autoradiography using the specific ligand 2-[125I]-iodomelatonin. To compare directly the distribution of IMEL-binding sites and one of the functional zones of the nucleus, alternate serial coronal sections through the SCN were processed for autoradiography for IMEL and one of the following: ICC for ChTB-ir or Fos-ir, or ISH for AVP mRNA. Overall, the regional distribution of the various markers within the SCN was comparable in the two species. The retinorecipient (ChTB-ir) and photically-responsive (Fos-ir) zones of the SCN mapped together to the middle and caudal thirds of the nucleus, predominantly in its ventro-lateral division.(ABSTRACT TRUNCATED AT 250 WORDS)

The suprachiasmatic area in the female hamster projects to neurons containing estrogen receptors and GnRH.

This study investigated whether the circadian regulation of luteinizing hormone (LH) release may be through direct input of the suprachiasmatic nucleus (SCN) to estrogen receptor (ER)- and/or gonadotropin releasing hormone (GnRH)-immunoreactive neurons. We used Phaseolus vulgaris leucoagglutinin (PHA-L) as an anterograde tracer of SCN efferents and performed double label immunocytochemistry for PHA-L and ER or GnRH. Between 8 and 30% of ER cells and 11-13% of the GnRH cells showed appositions with SCN efferents. Efferent projections of the subparaventricular hypothalamic nucleus and the retrochiasmatic area, relay stations of the circadian system, also made appositions with these two cell types. Results suggest that the circadian system could regulate the timing of the LH surge via two pathways, through input to GnRH and to ER cells.

Photoperiodic changes in opiate binding and their functional implications in golden hamsters.

Daylength modulates gonadotropin secretion, gonadal steroid hormone feedback, sexual behavior and body weight in male golden hamsters. Endogenous opiates regulate each of these phenomena, and the ability of opiate receptor blockade to elevate serum LH secretion is photoperiod-dependent. We used in vitro autoradiography to localize and quantify effects of daylength in golden hamsters. Hamsters were exposed to stimulatory (14 h light: 10 h dark) or inhibitory (10 h light: 14 h dark) photoperiods for 10 weeks before specific [3H]naloxone binding was assessed. Short days significantly decreased binding in medial amygdala and the intercalated amygdaloid nucleus. This effect was reversed by superior cervical ganglionectomy. No significant effects of daylength were observed in other amygdaloid, hypothalamic or preoptic areas. Lesions of the medial amygdala decreased copulatory behavior, short day-induced weight loss, and anogenital chemoinvestigation but did not affect gonadal regression or other forms of chemoinvestigation. These lesions facilitated testosterone's negative feedback on luteinizing hormone in long days but did not interfere with the potentiation of negative feedback by short days.

Melatonin binding sites in sciurid and hystricomorph rodents: studies on ground squirrels and guinea pigs.

Little is known about the distribution of binding sites for the pineal hormone melatonin in non-myomorph rodents. We used 2-[125I]iodomelatonin (IMEL) to analyze the distribution, affinity, and specificity of binding sites in the golden-mantled ground squirrel, a sciurid rodent that reportedly lacks IMEL binding sites in the brain. Specific binding was found not only in the pars tuberalis, but also in several telencephalic and diencephalic areas including the hypothalamic suprachiasmatic region. The affinity and specificity of IMEL binding are comparable to those reported in other rodents. IMEL binding studies in a hystricomorph rodent, the guinea pig, revealed high concentrations of receptor in the nucleus accumbens and dorsolateral thalamus. Central melatonin binding sites have now been demonstrated in species of all three rodent families. The heterogeneous distribution of melatonin receptors appears similar in the species studied, and no evidence is found to link IMEL binding sites at any particular locus to photoperiodic, circannual, or non-seasonal breeding patterns.