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Many species are threatened by climate change and must rapidly respond to survive changing environments. Epigenetic modifications, such as DNA methylation, can facilitate plastic responses by regulating gene expression in response to environmental cues. Understanding epigenetic responses is therefore essential for predicting species’ ability to rapidly adapt in the context of global environmental change. Here, we investigated the functional significance of DNA methylation on temperature-dependent life history in seed beetles, Callosobruchus maculatus. We assessed changes in DNA methyltransferase (Dnmt1 and Dnmt2) expression levels under ambient conditions and thermal stress, and reproductive performance following artificially-induced epimutation via 3-aminobenzamide (3AB) and Zebularine (Zeb), at a range of ambient and warmer temperatures over two generations. We found that Dnmt1 and Dnmt2 were greatly expressed in females, throughout the body, and exhibited temperature-dependence; in contrast, Dnmt expression was minimal in males. Epimutation led to shifts in female reproductive life history trade-off allocation, and differentially altered thermal optima of fecundity and offspring viability. This study revealed the optimal allocation strategy among these fitness components is temperature-dependent, and trade-offs become increasingly difficult to resolve epigenetically under more extreme warming. Results suggest that epigenetic mechanisms are strongly implicated in, and perhaps limiting of, invertebrate life history responses to temperature change. Further investigation will reveal targeted DNA methylation patterns and specific loci associated with temperature-dependent life history trade-offs in seed beetles and other invertebrates.

Seasonal changes in day length enhance and suppress immune function in a trait-specific manner. In Siberian hamsters (Phodopus sungorus) winter-like short days (SDs) increase blood leukocyte concentrations and adaptive T cell dependent immune responses, but attenuate innate inflammatory responses to simulated infections. Thyroid hormone (TH) signaling also changes seasonally and has been implicated in modulation of the reproductive axis by day length. Immunologically, TH administration in long days (LD) enhances adaptive immune responses in male Siberian hamsters, mimicking effects of SDs. This experiment tested the hypothesis that T3 is also sufficient to mimic the effects of SD on innate immune responses. Adult male hamsters housed in LDs were pretreated with triiodothyronine (T3; 1 μg, s.c.) or saline (VEH) daily for 6 weeks; additional positive controls were housed in SD and received VEH, after which cytokine, behavioral, and physiological responses to simulated bacterial infection (lipopolysaccharide; LPS) were evaluated. SD pretreatment inhibited proinflammatory cytokine mRNA expression (i.e. interleukin 1β, nuclear factor kappa-light-chain-enhancer of activated B cells). In addition, the magnitude and persistence of anorexic and cachectic responses to LPS were also lower in SD hamsters, and LPS-induced inhibition of nest building behavior was absent in SD. T3 treatments failed to affect behavioral (food intake, nest building) or somatic (body mass) responses to LPS in LD hamsters, but one CNS cytokine response to LPS (e.g., hypothalamic TNFα) was augmented by T3. Together these data implicate thyroid hormone signaling in select aspects of innate immune responses to seasonal changes in day length.

Many temperate zone animals engage in seasonal reproductive physiology and behavior as a strategy to maximise the propagation of the species. The hypothalamus integrates environmental cues and hormonal signalling to optimize the timing of reproduction. Recent work has revealed that epigenetic modifications, such as DNA methylation, vary across seasonal reproductive states. Multiple hormones act in the hypothalamus to permit or inhibit reproductive physiology, and the increase in thyroid hormone triiodothyronine (T3) has been implicated in the initiation of breeding in many species. The objective of this study was to examine the effect of T3 on the photoperiod-dependent regulation of reproductive physiology and hypothalamic DNA methyltransferase enzyme expression in female Siberian hamsters (Phodopus sungorus). We tested the hypothesis that T3 in short days (SD) would stimulate hypothalamic Rfrp3 and de novo DNA methyltransferase (Dnmt) expression in female Siberian hamsters. 10 weeks of SD lengths induced a decrease in body and uterine mass. Hamsters maintained in SD were found to express lower levels of GnRH, Rfrp3, Dnmt3a and Dnmt3b. Two weeks of daily T3 injections did not affect body mass, uterine mass, Gnrh, Rfrp3, Dnmt3a or Dnmt3b expression in neuroendocrine tissues. SD significantly lowered Tshβ mRNA expression and T3 reduced Tshβ in LD hamsters. Our data indicate sex-dependent effects of T3 for the neuroendocrine regulation of seasonal reproduction in hamsters.

Biomedical research is dominated by relatively few nonhuman animals to investigate healthy and disease conditions. Research has overrelied on these models due to their well-described genomes, the capability to control specific genes, and the high rate of reproduction. However, recent advances in large-scale molecular sequencing experiments have revealed, in some cases, the limited similarities in experimental outcomes observed in common rodents (i.e., mice) compared with humans. The value of more varied comparative animal models includes examples such as long-term body weight regulation in seasonally breeding hamsters as a means to help understand the obesity epidemic, vocal learning in songbirds to illuminate language acquisition and maintenance, and reproduction in cichlid fish to discover novel genes conserved in humans. Studying brain genes in prairie voles and cichlids advanced knowledge about social behavior. Taken together, experiments on diverse animal species highlight nontraditional systems for advancing our understanding of human health and well-being.

TLQP-21, a peptide encoded by the highly conserved vgf gene, is expressed in neuroendocrine cells and has been the most prominent VGF-derived peptide studied in relation to control of energy balance. The recent discovery that TLQP-21 is the natural agonist for the complement 3a receptor 1 (C3aR1) has revived interest in this peptide as a potential drug target for obesity. We have investigated its function in Siberian hamsters (Phodopus sungorus), a rodent that displays natural seasonal changes in body weight and adiposity as an adaptation to survive winter. We have previously shown that intracerebroventricular administration of TLQP-21 reduced food intake and body weight in hamsters in their long-day fat state. The aim of our current study was to determine the systemic actions of TLQP-21 on food intake, energy expenditure and body weight, and to establish whether adiposity affected these responses. Peripheral infusion of TLQP-21 (1mg/kg/day for 7 days) in lean hamsters exposed to short photoperiods (SP) reduced cumulative food intake in the home cage (p

Significance The genome and hypothalamic transcriptome of the Siberian hamster were sequenced and annotated to identify transcriptional pathways that exhibit seasonal plasticity in energy balance. Adaptation to short winter days reversed seasonal obesity and down-regulated hypothalamic proopiomelanocortin, and exogenous triiodothyronine reinstated weight gain and proopiomelanocortin expression. In silico analyses identified the evolution of thyroid hormone receptor binding motifs in the proximal promoter of the proopiomelanocortin gene of hamsters and other Cricetidae. Energetic challenges imposed by food restriction elicited orexigenic and anorexigenic neuropeptide responses in the hypothalamus, but did not affect proopiomelanocortin, which was regulated only by photoperiod. Hypothalamic proopiomelanocortin is maintained by photoperiod-driven triiodothyronine signaling and thereby affords adaptive long-term temporal organization of physiological systems that regulate energy balance., Synthesis of triiodothyronine (T3) in the hypothalamus induces marked seasonal neuromorphology changes across taxa. How species-specific responses to T3 signaling in the CNS drive annual changes in body weight and energy balance remains uncharacterized. These experiments sequenced and annotated the Siberian hamster (Phodopus sungorus) genome, a model organism for seasonal physiology research, to facilitate the dissection of T3-dependent molecular mechanisms that govern predictable, robust, and long-term changes in body weight. Examination of the Phodopus genome, in combination with transcriptome sequencing of the hamster diencephalon under winter and summer conditions, and in vivo-targeted expression analyses confirmed that proopiomelanocortin (pomc) is a primary genomic target for the long-term T3-dependent regulation of body weight. Further in silico analyses of pomc promoter sequences revealed that thyroid hormone receptor 1β-binding motif insertions have evolved in several genera of the Cricetidae family of rodents. Finally, experimental manipulation of food availability confirmed that hypothalamic pomc mRNA expression is dependent on longer-term photoperiod cues and is unresponsive to acute, short-term food availability. These observations suggest that species-specific responses to hypothalamic T3, driven in part by the receptor-binding motif insertions in some cricetid genomes, contribute critically to the long-term regulation of energy balance and the underlying physiological and behavioral adaptations associated with the seasonal organization of behavior.

It is becoming clear that epigenetic modifications such as DNA methylation can be dynamic and, in many cases, reversible. Here we investigated the photoperiod and hormone regulation of DNA methylation in testes, ovaries, and uterine tissue across multiple time scales. We hypothesized that DNA methyltransferase 3a (dnmt3a) is driven by photoperiodic treatment and exhibits natural variation across the female reproductive cycle and that melatonin increases whereas estrogen reduces DNA methylation. We used Siberian hamsters (Phodopus sungorus) due to their robust changes in reproductive physiology across seasonal and estrus time scales. Our findings indicate that short-day (SD) winter-like conditions significantly increased global DNA methylation and dnmt3a expression in the testes. Using immunohistochemistry, we confirm that increased dnmt3a expression was primarily localized to spermatogonium. Conversely, the ovaries did not exhibit variation in DNA methylation or dnmt3a/3b expression. However, exposure to SD significantly increased uterine dnmt3a expression. We then determined that dnmt3a was significantly decreased during the estrus stage. Next, we ovariectomized females and subsequently identified that a single estrogen+progesterone injection was sufficient to rapidly inhibit dnmt3a and dnmt3b expression. Finally, we demonstrate that treatment of human embryonic kidney-293 cells with melatonin significantly increased both dnmt3a and dnmt3b expression, suggesting that long-duration nocturnal signaling in SD may be involved in the regulation of DNA methylation in both sexes. Overall, our data indicate that dnmt3a shows marked photoperiod and estrus plasticity that likely has broad downstream effects on the timing of the genomic control of reproductive function.

Seasonal trade-offs in reproduction and immunity are ubiquitous in nature. The mechanisms that govern transitions across seasonal physiological states appear to involve reciprocal switches in the local synthesis of thyroid hormone. In long-day (LD) summer-like conditions, increased hypothalamic triiodothyronine (T3) stimulates gonadal development. Alternatively, short-day (SD) winter-like conditions increase peripheral leukocytes and enhance multiple aspects of immune function. These data indicate that the localized effects of T3 in the hypothalamus and leukocytes are photoperiod dependent. We tested the hypothesis that increased peripheral T3 in SD conditions would increase aspects of reproductive physiology and inhibit immune function, whereas T3 injections in LD conditions would facilitate aspects of immune function (i.e., leukocytes). In addition, we also examined whether T3 regulates hypothalamic neuropeptide expression as well as hypothalamic and splenic proinflammatory cytokine expression. Adult male Siberian hamsters were maintained in LD (15L:9D) or transferred to SD (9L:15D) for 8 weeks. A subset of LD and SD hamsters was treated daily with 5 µg T3 for 2 weeks. LD and SD controls were injected with saline. Daily T3 administration in SD hamsters (SD+T3) resulted in a rapid and substantial decrease in peripheral leukocyte concentrations and stimulated gonadal development. T3 treatment in LD (LD+T3) had no effect on testicular volumes but significantly increased leukocyte concentrations. Molecular analyses revealed that T3 stimulated interleukin 1β messenger RNA (mRNA) expression in the spleen and inhibited RFamide Related Peptide-3 mRNA expression in the hypothalamus. Moreover, there was a photoperiod-dependent decrease in splenic tumor necrosis factor–α mRNA expression. These findings reveal that T3 has tissue-specific and photoperiod-dependent regulation of seasonal rhythms in reproduction and immune function.

European starlings (Sturnus vulgaris) exhibit seasonal changes in singing and in the volumes of the neural substrate. Increases in song nuclei volume are mediated at least in part by increases in day length, which is also associated with increases in plasma testosterone (T), reproductive activity, and singing behavior in males. The correlations between photoperiod (i.e. daylength), T, reproductive state and singing hamper our ability to disentangle causal relationships. We investigated how photoperiodic-induced variation in reproductive state modulates the effects of T on singing behavior and song nuclei volumes in adult female starlings. Female Starlings do not naturally produce measureable levels of circulating T but nevertheless respond to exogenous T, which induces male-like singing. We manipulated photoperiod by placing birds in a photosensitive or photorefractory state and then treated them with T-filled or empty silastic implants. We recorded morning singing behavior for three weeks, after which we assessed reproductive condition and measured song nuclei volumes. We found that T-treated photosensitive birds sang significantly more than all other groups including T-treated photorefractory birds. All T-treated birds had larger song nuclei volumes than with blank-treated birds (despite photorefractory T-treated birds not increasing song-rate). There was no effect of photoperiod on the song nuclei volumes of T-treated birds. These data show that the behavioral effects of exogenous T can be modulated by reproductive state in adult female songbirds. Furthermore, these data are consistent with other observations that increases in singing rate in response to T are not necessarily due to the direct effects of T on song nuclei volume.

Precise timing of gene transcription is a fundamental component of many biological rhythms. DNA methylation and histone acetylation are two epigenetic modifications that can affect the probability of gene transcription and RNA expression. Enzymes involved in DNA methylation (dnmts) have been shown to exhibit photoperiodic rhythms in expression in the hypothalamus, which coincide with hypothalamic expression of deiodinase type III (dio3), a gene involved in the photoperiodic regulation of reproduction. It is currently unknown whether enzymes involved in histone deacetylation (hdacs) also vary in response to photoperiod, nor have seasonal changes in the circadian waveforms of methylation and/or acetylation enzymes been examined. The present work documents circadian and photoperiodic changes in dnmts and hdacs in whole hypothalamic dissections obtained from male Siberian hamsters (Phodopus sungorus) after 5–6 weeks of exposure to SD. The data indicate that short days (SD) markedly inhibit dnmt3a expression, and that SD inhibition of dnmt3a was evident regardless of the alignment of circadian waveforms. Among hdacs, photoperiodic and circadian changes in expression were only observed in hdac4 expression. Recurrent temporal waveforms in epigenetic enzyme expression may provide molecular inputs to the timing systems that reprogram RNA expression to generate daily and annual phenotypic plasticity.

Annual rhythms in morbidity and mortality are well-documented, and host defense mechanisms undergo marked seasonal phenotypic change. Siberian hamsters (Phodopus sungorus) exhibit striking immunological plasticity following adaptation to short winter day lengths (SD), including increases in blood leukocytes and in the magnitude of T cell-mediated immune responses. Thyroid hormone (TH) signaling is rate-limited by tissue-level expression of iodothyronine deiodinase types II and III (dio2, dio3), and dio2/dio3 expression in the central nervous system gate TH-dependent transduction of photoperiod information into the neuroendocrine system. THs are also potent immunomodulators, but their role in seasonal immunobiology remains unexamined. Here we report that photoperiod-driven changes in triiodothyronine (T3) signaling mediate seasonal changes in multiple aspects of immune function. Transfer from long days (LD) to SD inhibited leukocyte dio3 expression, which increased cellular T4→T3 catabolism. T3 was preferentially localized in the lymphocyte cytoplasm, consistent with a non-nuclear role of T3 in lymphoid cell differentiation and maturation. Exposure to SD upregulated leukocyte DNA methyltransferase expression and markedly increased DNA methylation in the dio3 proximal promoter region. Lastly, to bypass low endogenous T3 biosynthesis in LD lymphocytes, LD hamsters were treated with T3, which enhanced T cell-dependent delayed-type hypersensitivity inflammatory responses and blood leukocyte concentrations in a dose-dependent manner, mimicking effects of SD on these immunophenotypes. T3 signaling represents a novel mechanism by which environmental day length cues impact the immune system: changes in day length alter lymphoid cell T3-signaling via epigenetic transcriptional control of dio3 expression.

Annual changes in day length induce marked changes in reproductive function in temperate zone vertebrates. In many avian species, in contrast to other seasonally breeding animals, plasticity in hypothalamic gonadotropin-releasing hormone - 1 (GnRH1) expression rather than (or in addition to) release governs changes in pituitary-gonadal activity. Investigations of the cellular and molecular mechanisms that govern GnRH1 plasticity were previously hindered by a collective inability of scientists in the field to characterize the gnrh1 cDNA in songbirds. We finally overcame this roadblock after data from the zebra finch (Taeniopygia guttata) genome project enabled us to rapidly clone the gnrh1 cDNA from hypothalamic RNA of zebra finches and European starlings (Sturnus vulgaris). Here, we review the original data that identified GnRH1 protein plasticity in the songbird brain and discuss earlier failed attempts to clone gnrh1 in these animals. Then, we present recent efforts, including our own, that successfully characterized gnrh1 in zebra finch and starling, and demonstrated dynamic regulation of gnrh1 mRNA expression, particularly in sub-populations of preoptic area neurons, in the latter. Overall, this paper highlights GnRH1 plasticity in the avian brain, and weaves into the narrative the previously untold story of the challenges to sequencing gnrh1 in songbirds.

Sex differences in ultradian activity rhythms (URs) and circadian rhythms (CRs) were assessed in Siberian hamsters kept in long day (LD) or short day (SD) photoperiods for 40 weeks. For both sexes URs of locomotor activity were more prevalent, greater in amplitude and more robust in SDs. The UR period was longer in females than males in both day lengths. The reproductive system underwent regression and body mass declined during the initial 10 weeks of SD treatment, and in both sexes these traits spontaneously reverted to the LD phenotype at or before 40 weeks in SD, reflecting the development of neuroendocrine refractoriness to SD patterns of melatonin secretion. Hamsters of both sexes, however, continued to display SD-like URs at the 40 weeks time point. CRs were less prevalent and the waveform less robust and lower in amplitude in SDs than LDs; the SD circadian waveform also did not revert to the long-day phenotype after 40 weeks of SD treatment. Short day lengths enhanced ultradian and diminished circadian rhythms in both sexes. Day length controls several UR characteristics via gonadal steroid and melatonin-independent mechanisms. Sex differences in ultradian timing may contribute to sex diphenisms in rhythms of sleep, food intake and exercise.

Epigenetic modifications in reproductive tissues have predominantly focused on pathological conditions, such as ovarian and uterine cancers. The contribution of DNA methylation and histone acetylation to the timing and control of fertility is not well described. Siberian hamsters provide an important model to investigate the relatively short-term regulation of fertility (e.g. estrous) as well as long-term timing of breeding (e.g. seasonal). Recent work has shown that DNA methyltransferase 3a (dnmt3a) expression is associated with reproductive involution. Here, the objectives were to identify the impact of photoperiod on hdac1–3 expression in hamster testicular, ovarian and uterine tissue. Then, we assessed the effect of E2P4 and estrous cycling on hdac1–3 expression in uterine tissue. Testicular expression of hdac1 was significantly reduced, whereas hdac3 increased in reproductively photoregressed male hamsters; hdac2 expression did not significantly change across photoperiod conditions. There was no significant photoperiodic effect on ovarian expression of hdac1–3. Uterine expression of hdac3 expression was greater in long day hamsters; exposure to short days significantly reduced uterine hdac2 expression. Ovariectomized hamsters administered a single bolus injection of oil were found to have elevated uterine hdac2 compared to E2P4 treated females 12 h and 24 h post injection. Uterine hdac1–3 expression was relatively constant across the estrous cycle. Altogether these data indicate tissue-dependent photoperiodic regulation of hdac1–3 expression and that E2P4 may inhibit uterine hdac2 over long-term breeding cycles.

Seasonal breeding in temperate zone vertebrates is characterised by pronounced variation in both central and peripheral reproductive physiology as well as behaviour. In contrast, many tropical species have a comparatively longer and less of a seasonal pattern of breeding than their temperate zone counterparts. These extended, more “flexible” reproductive periods may be associate with a lesser degree of annual variation in reproductive physiology. Here we investigated variation in the neuroendocrine control of reproduction in relation to the changes in the neural song control system in a tropical breeding songbird the rufous-collared sparrows (Zonotrichia capensis). Using in situ hybridization, we show that the optical density of GnRH1 mRNA expression is relatively constant across pre-breeding and breeding states. However, males were found to have significantly greater expression compared to females regardless of breeding state. Both males and females showed marked variation in measures of peripheral reproductive physiology with greater gonadal volumes and concentrations of sex steroids in the blood (i.e. testosterone in males; estrogen in females) during the breeding season as compared to the pre-breeding season. These findings suggest that the environmental cues regulating breeding in a tropical breeding bird ultimately exert their effects on physiology at the level of the median eminence and regulate the release of GnRH1. In addition, histological analysis of the song control system HVC, RA and Area X revealed that breeding males had significantly larger volumes of these brain nuclei as compared to non-breeding males, breeding females, and non-breeding females. Females did not exhibit a significant difference in the size of song control regions across breeding states. Together, these data show a marked sex difference in the extent to which there is breeding-associated variation in reproductive physiology and brain plasticity that is dependent on the reproductive state in a tropical breeding songbird.

Gonadotropin-releasing hormone-I (GnRH-I) cells are localized primarily to the septopreoptic area (POA) and are responsible for regulating gonadotropin release from the anterior pituitary. Some songbird species exhibit dramatic seasonal variation in the number of detectable GnRH-I immunoreactive cells, with higher numbers being observed during the breeding season. Here we investigated the anatomical distribution of GnRH-I-immunoreactive cells in male starlings that varied in response to manipulations of reproductive state, social context, and gonadal condition. We housed photostimulated, intact and castrated male starlings with a female or alone. Additionally, a fifth treatment group consisted of photorefractory males (i.e., in a nonreproductive state) housed alone. All photostimulated males had significantly greater numbers of GnRH-I cells compared with photorefractory male starlings. There was a significant main effect of castration and social context. Castrated males had significantly greater numbers of GnRH-I cells compared with intact males, and males housed in male-female dyads also had greater numbers of GnRH-I cells. Furthermore, the significant main effects of castration and social context were the result of an increase in GnRH-I cell numbers specifically in the rostral and intermediate regions of the POA. These findings indicate that social context and hormonal milieu have profound effects on GnRH-I immunoreactivity in addition to the previously described effects of reproductive state. These data provide novel insight into the environmental regulation of the hypothalamopituitary axis and suggest that gonadal hormones and female presence independently regulate GnRH-I cells in specific regions of the POA in male starlings.

Many seasonally breeding avian species exhibit marked changes in hypothalamic content of gonadotropin-releasing vhormone 1 (GNRH1) protein that are reflective of breeding condition. We recently cloned the GNRH1 cDNA in European starlings and demonstrated that changes in GNRH1 mRNA levels occur with a time course similar to what has been observed with GNRH1 protein. However, we did not previously resolve whether these differences were attributable to changes in the number of cells expressing the gene. Herein, we investigated photoperiod-induced changes in the number and distribution of GNRH1 mRNA-expressing cells in the preoptic area of male starlings. GNRH1 mRNA-expressing cell number was significantly greater in breeding birds than in nonbreeding birds. Starlings maintained in short nonstimulatory day length (i.e., prebreeding) showed intermediate cell numbers. Detailed analysis of the rostrocaudal and mediolateral distribution revealed that breeding birds had greater numbers of cells expressing GNRH1 mRNA in the medial intermediate, mediocaudal, and lateral intermediate preoptic area compared with prebreeding and nonbreeding birds. These data demonstrate that photoperiodic changes in reproductive state in starlings are associated with region-specific alterations in the number of cells expressing the GNRH1 gene. It remains to be determined whether these changes reflect quantitative differences in gene expression among an otherwise stable population of cells or a phenotypic switch in which cells gain or lose the ability to make GNRH1 mRNA in response to environmental cues.

The effect of circadian rhythm (CR) disruption on immune function depends on the method by which CRs are disrupted. Behavioral and thermoregulatory responses induced by lipopolysaccharide (LPS) treatment were assessed in female Siberian hamsters in which circadian locomotor activity (LMA) rhythms were eliminated by exposure to a disruptive phase-shifting protocol (DPS) that sustains arrhythmicity even when hamsters are housed in a light-dark cycle. This noninvasive treatment avoids genome manipulations and neurological damage associated with other models of CR disruption. Circadian rhythmic (RHYTH) and arrhythmic (ARR) hamsters housed in a 16L:8D photocycle were injected with bacterial LPS near the onset of the light (zeitgeber time 1; ZT1) or dark (ZT16) phase. LPS injections at ZT16 and ZT1 elicited febrile responses in both RHYTH and ARR hamsters, but the effect was attenuated in the arrhythmic females. In ZT16, LPS inhibited LMA in the dark phase immediately after injection but not on subsequent nights in both chronotypes; in contrast, LPS at ZT1 elicited more enduring (~4 day) locomotor hypoactivity in ARR than in RHYTH hamsters. Power and period of dark-phase ultradian rhythms (URs) in LMA and Tb were markedly altered by LPS treatment, as was the power in the circadian waveform. Disrupted circadian rhythms in this model system attenuated responses to LPS in a trait- and ZT-specific manner; changes in UR period and power are novel components of the acute-phase response to infection that may affect energy conservation.

Production of T3 in the mediobasal hypothalamus is critical for regulation of seasonal reproductive physiology. Type 2 iodothyronine deiodinase (DIO2) and DIO3 enzymes catalyze the prohormone T4 into biologically-active T3 and biologically-inactive rT3, respectively. In several seasonally-breeding vertebrates, DIO2 and DIO3 expression is implicated in photoperiod signal transduction in adulthood. These experiments tested the hypothesis that juvenile Siberian hamsters, which are highly responsive to photoperiod at weaning (postnatal day [PND]18), exhibit rapid and sustained changes in hypothalamic dio3 mRNA expression during photoperiod-induced and photoperiod-inhibited puberty. Hypothalamic dio2 and dio3 expression was measured via quantitative PCR in hamsters born and reared in a long-day photoperiod (15L:9D) and weaned on PND18 into short-day photoperiods (9L:15D). In SD males, hypothalamic dio3 mRNA was elevated 2.5-fold within 3 days (PND21) and continued to increase (>20-fold) through PND32; changes in dio3 mRNA preceded inhibition of gonadotropin (FSH) secretion and gonadal regression in SD. Females exhibited comparable dio3 responses to SD. In LD males, dio3 remained low and invariant from PND18–PND32. In contrast, dio2 mRNA rose conspicuously on PND21, independent of photoperiod, returning to basal levels thereafter. In LD, a single afternoon melatonin (MEL) injection on PND18 or PND20 was sufficient to increase hypothalamic dio3 mRNA, and dio3 increased in proportion to the number of successive days of MEL treatment. SD photoperiods and MEL exert rapid, sustained, and additive effects on hypothalamic dio3 mRNA, which may play a central role in inhibiting maturation of the peripubertal hypothalamo-pituitary-gonadal axis.

The extent to which changes in ultradian and circadian rhythms (URs and CRs) reflect seasonal variations in pineal melatonin secretion was assessed in male Siberian hamsters transferred from long to short day lengths. The period of the locomotor activity UR increased from 2.5 h in long days to 4.5 h in short day lengths, but this and most other features of the short-day ultradian phenotype were unaffected by pinealectomy; only the short-day increase in UR amplitude was counteracted by pineal extirpation. Virtually all UR components were unaffected by gonadectomy or replacement testosterone or estradiol treatment; changes in testicular hormone secretion appear insufficient to account for seasonal fluctuation in URs. Pinealectomy did not affect activity onsets and offsets or phase angles of CR entrainment in short and long day lengths; the duration of nocturnal activity was equivalently longer in short than long days in both pinealectomized and pineal-intact hamsters. CR robustness of pinealectomized hamsters in short days was intermediate between values of long-day and short-day sham-pinealectomized males. Hourly nocturnal locomotor activity was markedly reduced in SD, and this effect was completely reversed by PINx. We conclude that seasonal transitions in UR and CR waveforms controlled by day length are mediated primarily by melatonin-independent mechanisms, with lesser contributions from melatonin-dependent processes. Most seasonal changes in ultradian and circadian rhythms in males of this species are not influenced by gonadal hormones. URs may allow animals to respond appropriately to changing environmental contingencies. In winter reduced activity combined with temporal restructuring of activity to include longer intervals of rest may be adaptive in maintaining body temperature at lower values and down-regulating energy expenditure when above ground temperatures are extremely low.

Estradiol facilitates the expression of male sexual behavior in Japanese quail within a few minutes. These rapid behavioral effects of estradiol could result from rapid changes in its local production in the preoptic area by aromatase, the enzyme converting testosterone into estradiol. Alternatively, aromatase activity may remain constant but fluctuations of local estradiol production could arise from rapid changes in the concentration of the enzymatic substrate, namely testosterone. Rapid increases of circulating testosterone levels have been observed in males of various species following social encounters. Surprisingly, in quail, the interaction with a female seems to result in a decrease in circulating testosterone levels. However, in that study conducted in quail, the samples were collected at intervals longer than the recently observed rapid effects of estradiol on sexual behavior. In the present study we investigated whether plasma testosterone concentrations fluctuate on a shorter time-frame. Eleven male were tested 5 min before and 5, 15 or 30 min after being allowed to have visual access to a female or to copulate with a female for 5 min. Both types of interactions resulted in a significant decline in circulating testosterone levels at latencies as short as 5 min. These data demonstrate that the decrease in testosterone levels is initiated shortly after sexual encounters. Because visual interactions with a female did not result in a rapid increase in testosterone concentrations, these findings rule out the possibility that a rapid rise in circulating testosterone levels participates in the rapid increase in brain estrogen synthesis and its facilitatory effects on copulatory behavior.

In songbirds, three isoforms of gonadotropin-releasing hormone (GnRH) have been reported: chicken GnRH-I (GnRH-I); and -II (GnRH-II); and lamprey GnRH-III (lGnRH-III). In galliformes, the distribution of GnRH-I and -II have been extensively studied. GnRH-I cell bodies are located primarily in the preoptic-septal region and primarily project to the median eminence. GnRH-II cell bodies are located in the midbrain and project to various hypothalamic and extrahypothalamic regions. In songbirds, the distributions of only GnRH-I and lGnRH-III have been thoroughly described. Songbirds differ from galliformes in that they possess specialized neural regions for the learning and production of complex vocalizations, the song-control system. In songbirds lGnRH-III was found within many song nuclei. In this study, we investigated the distribution of GnRH-II in the male house sparrow (Passer domesticus) brain. Our objectives were (1) to determine the distribution of GnRH-II in the male songbird brain and (2) to investigate the presence of GnRH-II in song control nuclei. Our study identified immunoreactive magnocellular GnRH-II cell bodies in the oculomotor region of the mesencephalon and parvicellular cells in the lateral hypothalamus. Also, an extensive network of GnRH-II immunoreactive fibres was observed in hypothalamic nuclei and extrahypothalamic regions. Immunoreactive GnRH-II fibres were most abundant in the preoptic area, ventromedial nucleus, hippocampus and the medial septum. Sparse immunoreactivity was also observed in the mesencephalon and diencephalon. GnRH-II was not identified in any of the song-control nuclei. Unlike lGnRH-III, these data suggest that GnRH-II is not directly involved with song learning, perception or production.

Background Production of contextually appropriate social behaviors involves integrated activity across many brain regions. Many songbird species produce complex vocalizations called ‘songs’ that serve to attract potential mates, defend territories, and/or maintain flock cohesion. There are a series of discrete interconnect brain regions that are essential for the successful production of song. The probability and intensity of singing behavior is influenced by the reproductive state. The objectives of this study were to examine the broad changes in gene expression in brain regions that control song production with a brain region that governs the reproductive state. Results We show using microarray cDNA analysis that two discrete brain systems that are both involved in governing singing behavior show markedly different gene expression profiles. We found that cortical and basal ganglia-like brain regions that control the socio-motor production of song in birds exhibit a categorical switch in gene expression that was dependent on their reproductive state. This pattern is in stark contrast to the pattern of expression observed in a hypothalamic brain region that governs the neuroendocrine control of reproduction. Subsequent gene ontology analysis revealed marked variation in the functional categories of active genes dependent on reproductive state and anatomical localization. HVC, one cortical-like structure, displayed significant gene expression changes associated with microtubule and neurofilament cytoskeleton organization, MAP kinase activity, and steroid hormone receptor complex activity. The transitions observed in the preoptic area, a nucleus that governs the motivation to engage in singing, exhibited variation in functional categories that included thyroid hormone receptor activity, epigenetic and angiogenetic processes. Conclusions These findings highlight the importance of considering the temporal patterns of gene expression across several brain regions when engaging in social behaviors.