Photic and nonphotic seasonal cues differentially engage hypothalamic kisspeptin and RFamide-related peptide mRNA expression in Siberian hamsters.

Seasonally breeding animals use a combination of photic (i.e. day length) and nonphotic (e.g. food availability, temperature) cues to regulate their reproduction. How these environmental cues are integrated is not understood. To assess the potential role of two candidate neuropeptides, kisspeptin and RFamide-related peptide-3 (RFRP), we monitored regional changes in their gene expression in a seasonally breeding mammal exposed to moderate changes in photoperiod and food availability. Adult male Siberian hamsters (Phodopus sungorus) were housed under a long (16 h light/day; 16 L) or intermediate (13.5 L) photoperiod and fed ad lib. or a progressive food restriction schedule (FR; reduced to 80% of ad lib.) for 11 weeks. Gonadal regression occurred only in FR hamsters housed under 13.5 L. Immunohistochemistry was used to identify diencephalic populations of kisspeptin- and RFRP-immunoreactive cells, and quantitative PCR was used to measure gene expression in adjacent coronal brain sections. Photoperiod, but not food availability, altered RFRP mRNA expression in the dorsomedial sections, whereas food availability but not photoperiod altered Kiss1 expression in the arcuate sections; intermediate photoperiods elevated RFRP expression, and food restriction suppressed Kiss1 expression. Regional- and neuropeptide-specific activity of RFamides may provide a mechanism for integration of multi-modal environmental information in the seasonal control of reproduction.

Affective and adrenocorticotrophic responses to photoperiod in Wistar rats.

The present study tested the hypothesis that seasonal intervals of exposure to modest changes in photoperiod, typical of those experienced by humans living in temperate latitudes (10-14 h light/day), engage changes in emotional behaviour of Wistar rats, a commonly-used animal model for investigations of affective physiology. Short day lengths (

Refractoriness to short day lengths augments tonic and gonadotrophin-releasing hormone-stimulated lutenising hormone secretion.

Siberian hamsters (Phodopus sungorus) undergo reproductive involution following exposure to short winter day lengths. Following approximately 20 weeks of exposure to short day (SD) lengths, hamsters become refractory to the inhibitory effects of SD, and reproductive competence is restored in anticipation of spring. The extent to which changes in gonadal steroid-dependent and -independent regulation of gonadotrophin secretion participate in this vernal reactivation of the gonads is not known. This experiment tested whether tonic and gonadotrophin-releasing hormone (GnRH)-stimulated regulation of lutenising hormone (LH) secretion differs between photoresponsive and photorefractory Siberian hamsters. Male hamsters born into long day (LD) lengths were castrated or subjected to a sham-castration surgery at 17 days of age, implanted s.c. with blank or testosterone-filled capsules, and housed in LD or SD thereafter. Baseline LH and LH responses to GnRH (200 ng/kg, s.c) were measured at 14 (photoresponsive) and 40 (photorefractory) weeks of age. Despite lower circulating testosterone concentrations in gonadally regressed SD hamsters on week 14, tonic LH concentrations were comparable among all groups of gonad-intact hamsters on weeks 14 and 40; however, week 14 SD hamsters exhibited significantly higher GnRH-stimulated LH responses. Tonic LH concentrations were indistinguishable among all groups of castrated hamsters bearing empty implants on week 14, but prolonged exposure to LD led to a decrease in resting LH, whereas prolonged exposure to SD resulted in an increase in LH. In castrated hamsters bearing testosterone implants, baseline LH concentrations were comparable in all groups, but GnRH treatment resulted in significantly higher LH concentrations in photorefractory (week 40, SD) hamsters relative to all other groups. The data suggest that the development of photorefractoriness in Siberian hamsters is characterised by enhanced gonadal hormone-independent stimulation of LH secretion, and diminished sensitivity to inhibitory negative-feedback effects of testosterone on LH secretion. Decreases in responsiveness of gonadotrophin secretion to gonadal hormone negative feedback may contribute to the process of photorefractoriness and assist in maintaining the growth of reproductive organs during the process of gonadal recrudescence.

Short day lengths enhance skin immune responses in gonadectomised Siberian hamsters.

In Siberian hamsters and other photoperiodic rodents, exposure to short photoperiods simultaneously inhibits gonadal hormone secretion and enhances some measures of immune function. The present study tested whether gonadal hormones mediate the effects of short days on skin immune function (delayed-type hypersensitivity reactions) in male Siberian hamsters. The magnitude of delayed-type hypersensitivity reactions was greater in hamsters exposed to short days relative to those in long days. Comparable effects of photoperiod were obtained in castrated hamsters bearing empty or testosterone-filled implants. The data suggest that contemporary gonadal hormone secretion is neither necessary, nor sufficient to mediate the effects of short photoperiods on skin immune function.

Photoperiod alters the time course of brain cyclooxygenase-2 expression in Siberian hamsters.

Fever is initiated by activation of the arachidonic acid cascade and the biosynthesis of prostaglandins within the brain. Inducible cyclooxygenase (COX-2) is a rate-limiting enzyme in prostaglandin synthesis, and the number of blood vessels expressing COX-2 correlates with elevated body temperature following peripheral lipopolysaccharide (LPS). Despite its importance in host defense, fever is energetically expensive and we hypothesized that fever may be limited by available metabolic resources. During winter, when competing metabolic demands are constrained by low temperatures and food availability, it was predicted that fever duration would be reduced in seasonally breeding Siberian hamsters (Phodopus sungorus). We measured LPS-induced COX-2 expression in blood vessels of hamsters to test whether photoperiodic alterations in fever duration are centrally mediated, or whether they reflect changes in peripheral modulation of body temperature. Hamsters housed in long, 'summer-like' or short, 'winter-like' day lengths for 10 weeks were injected with LPS, and brains were collected 2, 4, or 8 h later. COX-2 expression was comparably increased in long- and short-day hamsters by 2 h and 4 h post-LPS; however, short-day hamsters exhibited significantly fewer COX-2-positive cells and blood vessels by 8 h post-LPS compared to long-day hamsters, corresponding with reduced fever duration in short-day hamsters. Cortisol concentrations increased more than two-fold in short-day compared to long-day hamsters by 4 h; this increase may have contributed to the decrease in COX-2 expression observed by 8 h in short days. We conclude that short photoperiods significantly altered the time course of central COX-2 protein expression in hamsters in a manner consistent with reduced fever duration.

Photorefractoriness of immune function in male Siberian hamsters (Phodopus sungorus).

Short days induce multiple changes in reproductive and immune function in Siberian hamsters. Short-day reproductive inhibition in this species is regulated by an endogenous timing mechanism; after approximately 20 weeks in short days, neuroendocrine refractoriness to short-day patterns of melatonin develops, triggering spontaneous recrudescence of the reproductive system. It is unknown whether analogous mechanisms control immune function, or if photoperiodic changes in immune function are masked by prevailing photoperiod. In Experiment 1, 3 weeks of exposure to long days was not sufficient to induce long-day-like enhancement of in vitro lymphocyte proliferation in short-day adapted male Siberian hamsters. Experiment 2 tested the hypothesis that immunological photorefractoriness is induced by prolonged exposure to short days. Adult male hamsters were gonadectomized or sham-gonadectomized and housed in long (14 h light/day) or short (10 h light/day) photoperiods for 12, 32 or 40 weeks. Somatic and reproductive regression occurred after 12 weeks in short days, and spontaneous recrudescence was complete after 32-40 weeks in short days, indicative of somatic and reproductive photorefractoriness. In gonad-intact hamsters, 12 weeks of exposure to short days decreased the number of circulating granulocytes and increased the number of B-like lymphocytes. After 32 weeks in short days, these measures were restored to long-day values, indicative of photorefractoriness; castration eliminated these effects of photoperiod. In both intact and castrated hamsters, in vitro proliferation of splenic lymphocytes was inhibited by 12 weeks of exposure to short days. After 40 weeks in short days lymphocyte proliferation was restored to long-day values in intact hamsters, but remained suppressed in castrated hamsters. These results suggest that short-day-induced inhibition of lymphocyte function does not depend on gonadal regression, but that spontaneous recrudescence of this measure is dependent on gonadal recrudescence. In Experiment 3, in vitro treatment with melatonin enhanced basal proliferation of lymphocytes from male hamsters exposed to short days for 12 weeks, but had no effect on lymphocytes of photorefractory hamsters or long-day control hamsters. Lymphocytes of castrated hamsters were unresponsive to in vitro melatonin, suggesting that photoperiodic changes in gonadal hormone secretion may be required to activate mechanisms which permit differential responsiveness to melatonin depending on phase in the annual reproductive cycle. Together, these data indicate that, similar to the reproductive system, the immune system of male Siberian hamsters exhibits refractoriness to short days.

Spontaneous "regression" of enhanced immune function in a photoperiodic rodent Peromyscus maniculatus.

Short days inhibit reproduction and enhance immune function in deer mice (Peromyscus maniculatus). Their reproductive inhibition is sustained by an endogenous timing mechanism: after ca. 20 weeks in short days, reproductive photorefractoriness develops, followed by spontaneous recrudescence of the reproductive system. It is unknown whether analogous seasonal timing mechanisms regulate their immune function or whether enhanced immune function is sustained indefinitely under short days. In order to test this hypothesis, we housed adult male deer mice under long (16 h light day(-1)) or short (8 h light day(-1)) day conditions for 32 weeks or under long day conditions for 20 weeks followed by 12 weeks of short days. Mice under the long day conditions remained photostimulated over the 32 weeks, whereas mice housed under the short day conditions exhibited gonadal regression followed by photorefractoriness and spontaneous recrudescence. Mice transferred to short days at week 20 were reproductively photoregressed at week 32. Total splenocytes, relative splenic mass and mitogen-activated splenocyte proliferation were greater in those mice transferred to short days at week 20 than in those mice housed under either long or short day conditions for 32 consecutive weeks, and immune function in mice exposed to short days for 32 weeks was comparable with that of long day animals. These data suggest that short day enhancement of immune function is not indefinite. With prolonged (< or = 32 weeks) exposure to short days, several measures of immune function exhibit "spontaneous" regression, restoring long day-like immunocompetence. The results suggest that formal similarities and, possibly, common substrates exist among the photoperiodic timekeeping mechanisms that regulate seasonal transitions in reproductive and immune function.

Photoperiodic polyphenisms in rodents: neuroendocrine mechanisms, costs, and functions.

Annual changes in daylength figure prominently in the generation of seasonal rhythms in reproduction, and a wide variety of mammals use ambient photoperiod as a proximate cue to time critical reproductive events. Nevertheless, within many reproductively photoperiodic mammalian species, there exist individuals--termed "photoperiod nonresponders"--that fail to adopt a seasonal breeding strategy and instead exhibit reproductive competence at a time of year when their conspecifics are reproductively quiescent. Photoperiod nonresponsiveness has been principally characterized by laboratory observations--over half of the species known to be reproductively photoperiodic contain a proportion of nonresponsive individuals. The study of nonresponders has generated basic insights regarding photic regulation of reproduction in mammals. The neuroendocrine mechanisms by which the short-day photoperiodic signal is degraded or lost in nonresponders varies between species: differences in features of the circadian pacemaker, which provides photoperiodic input to the reproductive neuroendocrine system, have been identified in hamsters; changes in the responsiveness of hypothalamic gonadotrophs to melatonin and as-yet-unspecified inhibitory signals have been implicated in voles and mice. Individuals that continue to breed when their conspecifics refrain might enjoy higher fitness under certain circumstances. Statements regarding the adaptive function of reproductive nonresponsiveness to photoperiod require additional information on the costs (metabolic and fitness) of sustaining reproductive function during the winter months and how these costs vary as a function of environmental conditions. Reproductive nonresponders thus continue to represent a challenge to theories that extol the adaptive function of seasonality. Several nonexclusive hypotheses are proposed to account for the maintenance of nonresponsive individuals in wild rodent populations.

Photoperiod modulates the inhibitory effect of in vitro melatonin on lymphocyte proliferation in female Siberian hamsters.

In Siberian hamsters (Phodopus sungorus), short days suppress reproductive function and lymphocyte proliferation. To determine whether melatonin influences cell-mediated immunity through a direct action on lymphocyte proliferation, in vitro responsiveness to mitogens and melatonin was assessed in systemic and splenic lymphocytes from adult female Siberian hamsters housed in either long or short days for 13 weeks. Short days provoked reproductive regression and reduced lymphocyte proliferation. Physiological concentrations of melatonin (50 pg/ml) inhibited in vitro proliferation of circulating lymphocytes, whereas higher concentrations (> or = 500 pg/ml) were required to inhibit proliferation of splenic lymphocytes. Immunomodulatory effects of melatonin were restricted to lymphocytes from long-day hamsters-in vitro melatonin had no effect on circulating or splenic lymphocytes from females in short days. Responsiveness to melatonin in short-day lymphocytes may be restrained by the already expanded nightly pattern of melatonin secretion in short days. These data support the hypothesis that melatonin acts directly on lymphocytes from long-day hamsters to suppress blastogenesis.

Testicular development in Siberian hamsters depends on frequency and pattern of melatonin signals.

We investigated the impact of frequency and pattern of melatonin signals on reproductive development in Siberian hamsters. Juvenile males gestated in short day lengths and housed in constant illumination to suppress melatonin secretion were infused with melatonin for 5 h either once or twice per day for 20 days. Melatonin infusions at either frequency produced equivalent increases in testes and body weights that exceeded those of animals infused with saline but were indistinguishable from those of hamsters transferred to long day lengths. The reproductive system appears to be maximally stimulated by a single short melatonin signal each day. Other animals kept from birth in a short photoperiod were treated 6 h after onset of darkness with the beta-adrenergic receptor antagonist DL-propranolol to shorten melatonin secretion on the night of injection but not on subsequent nights. This permitted interpolation of short nightly melatonin signals of 4-5 h duration against a background of long melatonin signals of 10-12 h duration on other nights. Treatment regimes that maintained a 1:1 ratio of short to long melatonin signals for 8 wk stimulated reproductive development; a 1:2 signal ratio, in each of three different patterns, was uniformly ineffective. The number of successive short melatonin signals had little influence on the interval across which successive melatonin signals were summated to influence photoperiodic traits. The neuroendocrine axis appears more responsive to short melatonin signal frequency than pattern for development of the summer phenotype.

Establishment and persistence of photoperiodic memory in hamsters.

Long summer days unequivocally stimulate, and short winter days inhibit reproduction in Siberian hamsters. By contrast, intermediate-duration day lengths (12.5-14 h long) either accelerate reproductive development or initiate regression of the reproductive apparatus. Which of these outcomes transpires depends on an animal's photoperiodic history, suggesting that hamsters must encode a representation of prior photoperiods. The duration of nocturnal melatonin secretion is the endocrine representation of day length, but nothing is known about how long it takes to establish photoperiodic histories or how long they endure. Hamsters exposed for 2 or more weeks to long summer day lengths acquired a long-day photoperiodic history that determined subsequent reproductive responses to intermediate-duration day lengths and melatonin signals. The memory for long-day lengths persisted in pinealectomized hamsters for 6.5 weeks, faded significantly after 13 weeks, and was functionally absent after 20 weeks. These findings indicate that hamsters are influenced only by relatively recent day lengths and melatonin signals and ignore earlier ones that might cause them to misinterpret the salience of current day lengths.

Triggering of neuroendocrine refractoriness to short-day patterns of melatonin in Siberian hamsters.

Short day lengths induce gonadal regression within 6 weeks in Siberian hamsters, but this inhibitory effect eventually wanes and reproductive competence is regained after 4-6 months in short days. These experiments were undertaken to determine whether continuous exposure to short days for several months is necessary to induce neuroendocrine refractoriness, or if a few weeks of short days are sufficient to trigger processes that culminate in refractoriness several months later. Adult male Siberian hamsters transferred from their natal long-day photoperiod of 15L (15 h light/day) to a short-day photoperiod of 10L were pinealectomized (PINx) after 0, 6, 12, 21 or 40 weeks of exposure to 10L. Intact hamsters kept in 10L manifested gonadal regression by week 6 and completed gonadal recrudescence by week 40, at which time they too were PINx. Beginning at week 40 all hamsters were infused s.c. with melatonin for 10 h/day for 6 consecutive weeks. This treatment induces gonadal regression in photosensitive hamsters. At the end of melatonin treatment, maximal gonadal regression was recorded for hamsters PINx at week 0 and those PINx after 40 weeks in long days. Hamsters PINx after 40 week of short day treatment were completely unresponsive to melatonin whereas those PINx after 6 and 12 weeks had intermediate responses. The percentage of hamsters whose reproductive apparatus was refractory to melatonin at week 40 increased with increasing duration of exposure to short days prior to PINx. Refractoriness was induced by relatively few weeks of short days in some hamsters, whereas others required much more extensive exposure. Induction of refractoriness is triggered by a fraction of the short days that hamsters experience in nature but may not be manifested until many weeks later when it coincides with gonadal recrudescence. In nature all hamsters are exposed to short days for at least 25 weeks, ensuring refractoriness in most individuals.

Frequency coding of melatonin signals sufficient to induce testicular growth in photoregressed Siberian hamsters.

Integration of melatonin signals over the course of several weeks was studied in male Siberian hamsters maintained in a short day length (10 h light/day) from birth (d 0). On d 35, hamsters with undeveloped testes were housed in constant light and received 5 h s.c. infusions of melatonin (MEL; 100 ng/infusion) or saline (SAL) over the next 30 days. Infusions were provided either every day (30 MEL), every other day (1 MEL/1 OFF), for 2 consecutive days followed by 2 days with no infusion (2 MEL/2 OFF), for 5 consecutive days followed by 5 days with no infusion (5 MEL/5 OFF), for 15 consecutive days followed by 15 days with no infusion (15 MEL/15 OFF), or saline every day (30 SAL). On d 65 testes weights of 30 MEL hamsters were greater than those from 30 SAL, 1 MEL/1 OFF, or 2 MEL/2 OFF groups, but did not differ significantly from those of 5 MEL/5 OFF and 15 MEL/15 OFF animals. Serum FSH concentrations of 30 MEL hamsters exceeded those of all other groups which did not differ among each other. Between 15 and 30 consecutive daily melatonin signals are necessary and sufficient to initiate and sustain maximal gonadotropic activity in juvenile male hamsters kept in constant light. The neuroendocrine system responsive to melatonin does not bridge intervals of more than a day in any of several combinations and apparently is frequency-coded for maximal responsiveness to daily signals.

Pineal-independent regulation of photo-nonresponsiveness in the Siberian hamster (Phodopus sungorus).

The pineal hormone melatonin influences circadian rhythms and also mediates reproductive responses to photoperiod. The authors tested whether pinealectomy influences circadian oscillators responsible for induction of nonresponsiveness to short day lengths by preventing normal short-day patterns of circadian entrainment. Adult male Siberian hamsters were pinealectomized or sham operated, maintained in either 18 h light per day (18L) or 15L for 10 weeks, and then tested for responsiveness to 10L. Because pinealectomized hamsters do not show gonadal regression in short day lengths, responsiveness was assessed by measuring phase angle of entrainment and the length of the nightly activity period following transfer to 10L. The incidence of nonresponsiveness was significantly higher in 18L hamsters than in 15L hamsters but was unaffected by pineal status. Fully 88% of 18L hamsters failed to entrain to 10L in the normal short-day manner; the duration of nightly activity remained compressed, and the phase angle of entrainment was large and negative relative to lights off. The 15L hamsters entrained normally to 10L. Exposure to constant light after 10L treatment was equally effective in inducing arrhythmicity in pinealectomized and intact hamsters. Changes in the period of morning and evening circadian oscillators subsequent to 18L treatment did not predict circadian responsiveness to short photoperiod. Long-day induction of photo-nonresponsiveness, which prevents winter responses to short day lengths, occurs independently of pineal melatonin feedback on the circadian system.

Melatonin chimeras alter reproductive development and photorefractoriness in Siberian hamsters.

Nightly melatonin (MEL) durations > 8 h provoke gonadal regression and decreases in body mass, whereas signals < 7 h stimulate gonadal and somatic growth in male Siberian hamsters. The authors sought to determine the minimum frequency of short MEL signals sufficient to induce the long-day phenotype in several photoperiodic traits. D,L-propranolol (hereafter propranolol) injections shortened MEL signals on the night of treatment without altering MEL on the subsequent night; this permitted interpolation of short MEL signals at variable frequencies against a background of long MEL signals (chimeras). Hamsters kept in short days (10 h light/day, 10L) were injected with propranolol 6 h after dark onset for 28 consecutive weeks beginning at 30 days of age (Week 0) either every other day or once every 3, 6, or 9 days. Control animals were injected with saline or with propranolol during the light phase or were transferred to long days (16L) at Week 0. Hamsters in 16L underwent rapid gonadal development and increases in body mass and displayed summer pelage color, as did hamsters treated with propranolol every other day. Animals treated with propranolol less frequently than every other day uniformly maintained undeveloped gonads and winter-like body weights, but pelage color became proportionately darker with increased frequency of propranolol treatments. The onset of spontaneous testicular development in 10L was unaffected by propranolol injections. After termination of injections at Week 28, testicular regression was not observed in most 10L animals that previously had undergone spontaneous testicular development; however, 40% of hamsters that had been injected with propranolol every 3rd night did manifest the winter phenotype after Week 28. In an alternating sequence, short MEL signals completely override long signals and induce the summer phenotype. Threshold frequencies differ for MEL stimulation of long-day pelage and gonadal phenotypes. The timing and development of refractoriness to MEL does not depend in any simple manner on the number of long MEL signals or on the accumulation of a reaction product produced by long, and depleted by short, MEL signals.

Enhanced reproductive responses to melatonin in juvenile Siberian hamsters.

Reproductive responses to melatonin (Mel) exposure were assessed in male Siberian hamsters gestated in long (16 h light/day; 16L) or short (8 h light/day; 8L) day lengths (DLs). Hamsters were maintained in constant light beginning at 14 days of age (day 14) and were infused for 3 successive days beginning on day 18, 25, or 32 with long (12 h/day) or short (6 h/day) durations of Mel or saline. Two weeks later, testis and body weights were determined. Responsiveness to Mel was enhanced in hamsters beginning shortly after weaning and was influenced by photoperiodic history. Gonadal growth in 16L hamsters was inhibited by 12-h Mel infusions initiated on day 18 or day 25, but infusions initiated on day 32 were ineffective. Hamsters gestated in 8L were not responsive to Mel treatments initiated on day 18, but discriminated 12-h from 6-h infusions at day 25 and manifested reduced gonadal growth after 12-h Mel infusions on day 32. Hamsters born in 8L to dams judged as photoinsensitive to short DLs did not respond to 12-h infusions on day 32 as inhibitory signals. We propose that maternal Mel secretion provides juvenile males with a photoperiodic history, which allows them to compare gestational with early postweaning Mel signals and thereby program seasonally appropriate developmental trajectories.

Comparative studies on the Mauthner cell of teleost fish in relation to sensory input.

Most physiological and behavioral studies of the Mauthner cells have used the goldfish and a few other fish from the superorder Ostariophysi, series Otophysi (= otophysans). We first provide some background and recent findings on the Mauthner cells of otophysan fish and then compare this information to that known about the Mauthner cells in certain non-otophysan fish. These comparisons are meant to provide the impetus for a comparative approach to understanding the role of the Mauthner cells in behavior.