Disrupted seasonal biology impacts health, food security and ecosystems.

The rhythm of life on earth is shaped by seasonal changes in the environment. Plants and animals show profound annual cycles in physiology, health, morphology, behaviour and demography in response to environmental cues. Seasonal biology impacts ecosystems and agriculture, with consequences for humans and biodiversity. Human populations show robust annual rhythms in health and well-being, and the birth month can have lasting effects that persist throughout life. This review emphasizes the need for a better understanding of seasonal biology against the backdrop of its rapidly progressing disruption through climate change, human lifestyles and other anthropogenic impact. Climate change is modifying annual rhythms to which numerous organisms have adapted, with potential consequences for industries relating to health, ecosystems and food security. Disconcertingly, human lifestyles under artificial conditions of eternal summer provide the most extreme example for disconnect from natural seasons, making humans vulnerable to increased morbidity and mortality. In this review, we introduce scenarios of seasonal disruption, highlight key aspects of seasonal biology and summarize from biomedical, anthropological, veterinary, agricultural and environmental perspectives the recent evidence for seasonal desynchronization between environmental factors and internal rhythms. Because annual rhythms are pervasive across biological systems, they provide a common framework for trans-disciplinary research.

Egr1 involvement in evening gene regulation by melatonin.

Seasonal photoperiodic responses in mammals depend on the pineal hormone melatonin. The pars tuberalis (PT) region of the anterior pituitary has emerged as a principal melatonin target tissue, controlling endocrine responses. Rising melatonin levels acutely influence the expression of a small cluster of genes either positively (exemplified by cryptochrome-1, cry1) or negatively (exemplified by the type 1 melatonin receptor, mt1). The purpose of this study was to characterize the pathways through which these evening actions of melatonin are mediated. In vitro experiments showed that cAMP signaling in the PT directly influences mt1 but not cry1 expression. Analysis of nuclear extracts from sheep PT tissue collected 90 min after melatonin or saline control injections highlighted the response element for the immediate early gene egr1 (EGR1-RE) as a candidate for acute melatonin-dependent transcriptional regulation. We identified putative EGR1-RE's in the proximal promoter regions of the ovine cry1 and mt1 genes, and confirmed their functionality in luciferase reporter assays. Egr1 expression is suppressed by melatonin in PT cell cultures, and is rhythmic in the ovine PT with a nadir in the early night. We propose that melatonin-dependent effects on EGR1-RE's contribute to evening gene expression profiles in this pituitary melatonin target tissue.

Energy conservation.

We can no longer afford to ignore the serious potential consequences of our lavish use of energy. Continuation of the present rate of increase, particularly with the trend to imported fuels, will lead in short order to a level of dependency on imports which is disturbing for both the national security and the balance of payments. The inevitable rise in the price of energy will presumably lead to some increases in the domestic energy supply. But our reserves, particularly in the preferred forms of petroleum, gas, and even low-sulfur coal, are finite. Thus, the energy problem must also be attacked from the standpoint of energy conservation. The forthcoming rise in fuel prices will, of course, make more attractive some forms of conservation which at present are economically marginal. Nevertheless, consumers, industry, and government will have to make difficult choices in the years ahead: between greater convenience and lower energy bills, between the high capital costs of energy conservation measures and the long-term dollar savings from increased energy efficiency, and between environmental protection and the availability of needed energy supplies. Existing capabilities and technology, on which short- and midterm improvements must be based, appear to offer substantial possibilities for reducing U.S. energy consumption within the next decade (11). Long-term solutions to the energy problem, however, will depend to a considerable extent on the continuing appearance of new technological capabilities for increased efficiency of energy utilization and increased integration of energy applications. The capacity for continuing technological advances is, of course, dependent in turn on a strong relevant scientific base. A word of caution is necessary. Recent experience has shown that technological advances alone will not solve the problem. The problem spans not only the traditional physical and engineering sciences but also those sciences which deal with human attitudes and actions, that is, the social sciences, and includes a more fundamental understanding of underlying economic principles. The challenge to all sectors of American science should be clear.

Phase shifts in the circadian rhythm in plasma concentrations of melatonin in rams induced by a 1-hour light pulse.

The effect of a 1-hr light pulse, given at night, on the timing of the circadian rhythm in the plasma concentration of melatonin was examined in Soay rams to investigate the mechanisms involved in determining the duration of the nocturnal peak in melatonin secretion. Animals (n = 8) were housed under short days (LD 8:16) or long days (LD 16:8) and received a light pulse at various times of night. They were released into constant dim red light (DD) on day 1. Blood samples were collected hourly for 30 hr from 1000 hr on day 3, and the plasma concentration of melatonin was determined by radioimmunoassay to assess the timing of the melatonin peak. Control animals (n = 8) were maintained under the same conditions but received no light pulse. Under short days, a light pulse given early in the night caused a phase delay in the melatonin peak, and a light pulse given in the late night caused a phase advance. The mean duration of the melatonin peak was slightly reduced following a light pulse in the early or late night, and slightly increased following a pulse given near the middle of the night. Under long days, both light-pulse treatments given at night caused a phase delay in the melatonin peak, but there was no significant change in duration of the melatonin peak. The duration of the melatonin peak at day 3 under DD in the control animals was similar for all treatments, regardless of the previous entraining photoperiod (mean duration: 12.6-14.8 hr) and was similar to that under short days (14.6 hr), but was significantly longer than that under long days (8.2 hr). Information on the phase response curve in the Soay ram and on the period of the circadian oscillator governing the melatonin rhythm (c 23.0 hr under DD) predicts a close phase relationship between the end of the light phase and the onset of the melatonin peak as observed under normal 24-hr LD cycles. The current results also indicate that light acts to entrain the circadian rhythm influencing the onset and offset of melatonin secretion, and thus dictates the duration of the melatonin peak.

Light control of the duration of the daily melatonin signal under long and short days in the Soay ram. Role of inhibition and entrainment.

Light acts in two ways to control the duration of the nocturnal melatonin rhythm. It inhibits the production of melatonin from the pineal gland and it entrains the underlying circadian rhythm generators located in the suprachiasmatic nuclei. To investigate the role of these two mechanisms under long and short days, four experiments were carried out using groups of adult Soay rams (n = 6-8). The animals were housed in individual pens in light-controlled rooms and entrained to long (LD 16:8) or short (LD 8:16) days for at least 8 wk. The treatments were as follows: (i) dark period extended by 4 h under long days (L dark-delay), (ii) dark period advanced by 4 h under long days (L dark-advance), (iii) dark period extended by 4 h under short days (S dark-delay), and (iv) dark period advanced by 4 h under short days (S dark-advance). Each treatment was given on a single day and the animals were subsequently maintained in, or transferred to, constant dim red light (DD) for 24 h. A control group (C) was run in parallel with each treatment group. Blood samples were collected every 30 min for 6-9 h during the dark-shift to monitor the light-induced changes in the secretion of melatonin, and during DD to monitor any phase shift in the endogenous rhythm (phase markers provided by onset or offset of melatonin secretion). L dark-delay resulted in a significantly (p < 0.01, ANOVA) later offset of the melatonin peak (3.4-h delay) with no phase shift of the onset of the rhythm under DD.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of constant darkness and constant light on circadian organization and reproductive responses in the ram.

The relationship between circadian rhythms in the blood plasma concentrations of melatonin and rhythms in locomotor activity was studied in adult male sheep (Soay rams) exposed to 16-week periods of short days (8 hr of light and 16 hr of darkness; LD 8:16) or long days (LD 16:8) followed by 16-week periods of constant darkness (dim red light; DD) or constant light (LL). Under both LD 8:16 and LD 16:8, there was a clearly defined 24-hr rhythm in plasma concentrations of melatonin, with high levels throughout the dark phase. Periodogram analysis revealed a 24-hr rhythm in locomotor activity under LD 8:16 and LD 16:8. The main bouts of activity occurred during the light phase. A change from LD 8:16 to LD 16:8 resulted in a decrease in the duration of elevated melatonin secretion (melatonin peak) and an increase in the duration of activity corresponding to the changes in the ratio of light to darkness. In all rams, a significant circadian rhythm of activity persisted over the first 2 weeks following transfer from an entraining photoperiod to DD, with a mean period of 23.77 hr. However, the activity rhythms subsequently became disorganized, as did the 24-hr melatonin rhythms. The introduction of a 1-hr light pulse every 24 hr (LD 1:23) for 2 weeks after 8 weeks under DD reinduced a rhythm in both melatonin secretion and activity: the end of the 1-hr light period acted as the dusk signal, producing a normal temporal association of the two rhythms. Under LL, the 24-hr melatonin rhythms were disrupted, though several rams still showed periods of elevated melatonin secretion. Significant activity rhythms were either absent or a weak component occurred with a period of 24 hr. The introduction of a 1-hr dark period every 24 hr for 2 weeks after 8 weeks under LL (LD 23:1) failed to induce or entrain rhythms in either of the parameters. The occurrence of 24-hr activity rhythm in some rams under LL may indicate nonphotoperiodic entrainment signals in our experimental facility. Reproductive responses to the changes in photoperiod were also monitored. After pretreatment with LD 8:16, the rams were sexually active; exposure to LD 16:8, DD, or LL resulted in a decline in all measures of reproductive function. The decline was slower under DD than LD 16:8 or LL.(ABSTRACT TRUNCATED AT 400 WORDS)

Compensatory response of the luteinizing-hormone (LH)-releasing hormone (LHRH)/LH pulse generator after administration of a potent LHRH antagonist in the ram.

It is established that the blockade of the pituitary LHRH receptor by an LHRH antagonist will suppress pituitary LH secretion and reduce serum concentrations of gonadal steroids. Little is known, however, about the activity of the LHRH/LH pulse generator during this inhibitory period or during the recovery phase. To investigate this, a potent LHRH antagonist [N-Ac-D-pCl-Phe1,D-pCl-Phe2,D-Trp3,D-hArg(Et2)6, D-Ala10 LHRH was injected iv into sexually active rams and the changes in the blood plasma concentrations of LH, FSH, testosterone, and PRL were measured in samples collected every 15 min for 24-48 h. The treatment induced an immediate blockade of pulsatile LH secretion and a parallel decline in blood levels of testosterone. Plasma levels of FSH were not suppressed by treatment with the LHRH antagonist and there was no consistent effect on plasma levels of PRL. The duration of the inhibition of LH was dose dependent lasting 4.3 +/- 0.4 h, 18.0 +/- 1.0 h, and 31.8 +/- 1.3 h for the low (6 micrograms/kg), medium (36 micrograms/kg), and high (365 micrograms/kg) doses of LHRH antagonist, respectively. During the recovery period there was an approximate 2-fold increase in the frequency of LH pulses. These results suggest a compensatory response to the decline in the negative feedback effect of testosterone secretion. Even the lowest dose of antagonist elicited a decrease in the level of testosterone and an increase in LH pulse frequency. At this dose, the decline in testosterone was very transitory indicating an acute sensitivity of the hypothalamus to changes in the negative feedback signal. These results suggest that the suppression of LH and testosterone secretion in the ram by LHRH antagonist is associated with a compensatory increase in the activity of the LHRH pulse generator.

Beta-endorphin secretion in rams related to season and photoperiod.

A newly established RIA was used to measure changes in the concentration of beta-endorphin in peripheral blood and pituitary tissue from adult Soay rams living outside under natural conditions and housed indoors under artificial photoperiods. A pronounced seasonal cycle in plasma beta-endorphin immunoreactivity occurred in the outdoor animals, with low levels in spring and early summer (February-May; less than 200 pg/ml plasma) and maximal levels 10-20 times higher in late summer and autumn (July-October). Seasonal changes in plasma levels of PRL, FSH, and cortisol, testis size, and body weight were also monitored; the seasonal cycle in the levels of immunoreactive beta-endorphin occurred in parallel with the cycle in plasma FSH and body weight. There were no significant seasonal changes in plasma cortisol concentrations. Marked changes in the plasma levels of beta-endorphin were also seen in rams kept under the artificial photoperiod regimen of alternating 12- to 16-week periods of long days (16 h of light and 8 h of darkness; 16L:8D) and short days (8L:16D). Transfer from long days to short days led to a greater than 20-fold increase in the levels of beta-endorphin, reaching a maximum after 4-8 weeks; the reverse switch in photoperiod led to a rapid decrease in the levels. There was no diurnal rhythm in the plasma levels of beta-endorphin based on hourly samples collected for 24 h under long and short days. The total content of immunoreactive beta-endorphin in the pituitary gland was lower in rams under short days than under long days, converse to the pattern in the blood. Sephadex chromatography of the plasma samples revealed that most of the beta-endorphin immunoreactivity coeluted with synthetic beta-endorphin-(1-31), and a small amount of activity eluted with beta-lipotropin. The seasonal and photoperiod-induced changes were largely due to changes in the levels of beta-endorphin. Extracts of pituitary tissue revealed a large proportion of beta-lipotropin to beta-endorphin compared to plasma, with no consistent change in ratio related to the photoperiod. The overall results illustrate that there are pronounced seasonal and photoperiod-induced changes in immunoreactive plasma beta-endorphin levels in the ram. Under artificial photoperiods, long days inhibit and short days stimulate beta-endorphin secretion. Under natural conditions, the development of refractoriness to both the inhibitory effects of long days and the stimulatory effects of short days may explain the timing of the annual cycle of beta-endorphin secretion.

Photoperiodic control of thyroid function and wool and horn growth in rams and the effect of cranial sympathectomy.

Eight adult Soay rams (four control and four superior cervical ganglionectomized) were housed in an artificial lighting regimen of alternating 16-week periods of long days [16 h of light, 8 h of darkness (16 L:8D)] and short days (8L:16D) for nearly 3 yr, and the long term variations in growth of the wool and horns were recorded along with measurements of the plasma concentration of T4, T3, and testosterone. In the control rams all of the parameters varied in relation to the imposed lighting regimen. An increase in the growth of the wool and horns occurred during each period of long days at a time when the circulating concentrations of T4 and T3 were high; at this time the animals were reproductively quiescent, with low plasma levels of testosterone. During each period of short days the rate of wool and horn growth declined, as did the plasma concentrations of T4 and T3, while the rams became sexually active with high plasma levels of testosterone. Moulting of the old coat occurred during the periods of long days, coinciding with the phase of increasing wool growth. The variations in the ganglionectomized rams were quite different from those in the controls, bearing no clear relationship to the imposed lighting regimen. In these operated animals there were long term changes in all of the parameters, but these were generally less pronounced than those in the controls. The unusual pattern in the growth of horns and wool was correlated with long term changes in the plasma levels of T4, T3, and testosterone. Moulting of the old coat did not occur in the normal manner in the animals after the first year.

Cell type-specific adenoviral transgene expression in the intact ovine pituitary gland after stereotaxic delivery: an in vivo system for long-term multiple parameter evaluation of human pituitary gene therapy.

Ablative therapies for pituitary tumors commonly cause irreversible damage to normal pituitary cells. Toxin gene therapy should therefore ideally be targeted to specific cell types to avoid collateral cell damage. To evaluate cell-type-specific adenoviral gene transfer in the intact pituitary gland we have used stereotaxic transcranial delivery of recombinant adenoviruses in the sheep with continuous assessment of endocrine function. Adenoviral ss-galactosidase expression was driven either by the human cytomegalovirus (hCMV) promoter or the human PRL gene promoter. The hCMV promoter directed adenoviral ss-galactosidase expression in all pituitary cell types, but the PRL promoter restricted this exclusively to lactotropic cells, indicating that this promoter conferred appropriate cell type specificity in the context of adenoviral transduction in vivo. Serial measurements of plasma hormones showed no disruption of endocrine function over 7 days after intrapituitary injection. In summary, this work shows cell type-specific expression of an adenoviral transgene in the mixed cell population of the intact pituitary gland in vivo in a large animal model and indicates that stereotaxic intrapituitary delivery does not disrupt normal endocrine function.

Rhythmic melatonin secretion does not correlate with the expression of arylalkylamine N-acetyltransferase, inducible cyclic amp early repressor, period1 or cryptochrome1 mRNA in the sheep pineal.

The pineal gland, through nocturnal melatonin, acts as a neuroendocrine transducer of daily and seasonal time. Melatonin synthesis is driven by rhythmic activation of the rate-limiting enzyme, arylalkylamine N-acetyltransferase (AA-NAT). In ungulates, AA-NAT mRNA is constitutively high throughout the 24-h cycle, and melatonin production is primarily controlled through effects on AA-NAT enzyme activity; this is in contrast to dominant transcriptional control in rodents. To determine whether there has been a selective loss of circadian control of AA-NAT mRNA expression in the sheep pineal, we measured the expression of other genes known to be rhythmic in rodents (inducible cAMP early repressor ICER, the circadian clock genes Period1 and Cryptochrome1, as well as AA-NAT). We first assayed gene expression in pineal glands collected from Soay sheep adapted to short days (Light: dark, 8-h: 16-h), and killed at 4-h intervals through 24-h. We found no evidence for rhythmic expression of ICER, AA-NAT or Cryptochrome1 under these conditions, whilst Period1 showed a low amplitude rhythm of expression, with higher values during the dark period. In a second group of animals, lights out was delayed by 8-h during the final 24-h sampling period, a manipulation that causes an immediate shortening of the period of melatonin secretion. This did not significantly affect the expression of ICER, AA-NAT or Cryptochrome1 in the pineal, whilst a slight suppressive effect on overall Per1 levels was observed. The attenuated response to photoperiod change appears to be specific to the ovine pineal, as the first long day induced rapid changes of Period1 and ICER expression in the hypothalamic suprachiasmatic nuclei and pituitary pars tuberalis, respectively. Overall, our data suggest a general reduction of circadian control of transcript abundance in the ovine pineal gland, consistent with a marked evolutionary divergence in the mechanism regulating melatonin production between terrestrial ruminants and fossorial rodents.

Photoperiodic control of seasonal breeding in the ram: participation of the cranial sympathetic nervous system.

Four mature Soay rams, cranially sympathectomized by removal of the superior cervical ganglia, were housed alongside four normal rams controlled lighting conditions of alternating 16 week periods of short days of 8 h light: 16 h darkness (8L : 16D) and long days (16L : 8D). The changes in the concentration of FSH, LH, prolactin and testosterone in the plasma, the size of the testes, the intensity of the sexual flush and the sexual and aggressive behaviour of the animals were recorded. While the control rams were able to respond to the artificial lighting conditions with synchronized cycles of reproductive activity, the ganglionectomized animals failed to respond. The treated rams had well-developed testes and relatively high levels of gonadotrophins and testosterone in the blood throughout the experiment. It is concluded that the cranial sympathetic nervous system is involved in the photoperiodic control of seasonal breeding in the ram, probably through its role in the innervation of the pineal gland.

Effects of placing micro-implants of melatonin in the pars tuberalis, pars distalis and the lateral septum of the forebrain on the secretion of FSH and prolactin, and testicular size in rams.

Previous studies involving the placement of microimplants of melatonin in the brain in sheep exposed to long days have provided evidence that melatonin acts within or close to the mediobasal hypothalamus (MBH) to mediate the effects of daylength on cycles in reproduction, moulting and other seasonal characteristics. To extend these observations, groups of Soay rams have now been treated with micro-implants of melatonin placed in the pars tuberalis (PT) and pars distalis (PD) of the pituitary gland, and in the lateral septum of the forebrain (septum). The PT and septum are potential target sites for the action of melatonin based on the localized binding of iodomelatonin assessed by in situ autoradiography. The animals were initially exposed to alternating 16-week periods of long days (16 h light: 8 h darkness; 16L:8D) and short days (8L:16D) to entrain the seasonal cycles. The treatments were started at 10 weeks into a period of long days when the animals had a physiology normally observed in summer (low blood plasma concentrations of FSH and high concentrations of prolactin), and they remained under long days throughout the experiments. In experiment 1, animals received micro-implants of melatonin placed in the PT (n = 6) or PD (n = 4), or received empty implants in similar sites (n = 4) or no surgery (n = 4; total control, n = 8). In experiment 2, groups of animals received microimplants of melatonin placed in the lateral septum (septum, n = 7) or received corresponding control treatments (total control, n = 8). The micro-implants consisted of 22 gauge stainless-steel needles with melatonin fused inside the tip. They were inserted bilaterally in the selected sites and left in place for 14 weeks. The biological effects of the treatments were assessed by measuring the changes in the blood plasma concentrations of FSH and prolactin, growth of the teses and moulting of the pelage over a period of 28 weeks (14 weeks treatment and 14 weeks post-treatment). The administration of melatonin in the PT, but not in the PD or septum, affected the photoperiodically induced cycle in the secretion of FSH and prolactin. In the PT group there was no significant change in the plasma concentrations of FSH during the treatment with melatonin, but there was a significant (P < 0.001, ANOVA) decrease in the levels of FSH after the treatment associated with premature regression of the testes.(ABSTRACT TRUNCATED AT 250 WORDS)

Changes in pituitary responsiveness to luteinizing hormone releasing hormone in rams exposed to artificial photoperiods.

Six adult Soay rams were subjected to an artificial lighting régime of alternating 4 month periods of long days (16 h light:8 h darkness) and short days (8 h light:16 h darkness) which induced a seasonal cycle in testicular development and regression during a period of 32 weeks. On 15 occasions during the study pituitary responsiveness was assessed by measuring the concentration of luteinizing hormone (LH) in jugular blood samples collected at frequent intervals after the intravenous injection of 1 microgram synthetic luteinizing hormone releasing hormone (LH-RH). It was shown that both the magnitude and duration of the induced release of LH changed in relation to the photoperiod; the magnitude was greatest during long days when the rams were sexually quiescent, while the duration of the LH release was greatest during short days at the peak of testicular activity. This pattern of responsiveness was modified by implantation of the rams with testosterone.

Plasma testosterone profiles in male macropodid marsupials.

Serial blood samples were collected over a 3.5-8 h period from ten adult male macropodid marsupials belonging to five different species to study the pattern of testosterone secretion. The concentration of testosterone in the plasma ranged from 0.5 to 9.5 ng/ml, and in each animal the levels declined progressively during the study; this effect was attributed to the stress effects of handling. Injection of 5 microgram synthetic luteinizing hormone releasing hormone induced a rapid and substantial increase in the level of testosterone.

Seasonal variation in the episodic secretion of luteinizing hormone and testosterone in the ram.

Rams of an ancient breed of domestic sheep (Soay) were housed under artificial lighting conditions to study the way in which the secretion of LH and testosterone changes in relation to the mating season. Conspicuous changes were found in the short-term fluctuations in plasma LH concentrations related to the cycle of testis growth and regression; serial blood samples collected at short intervals revealed episodic peaks in plasma LH at all times, but there were changes in the frequency (lowest when the testes were regressed and highest when fully active), amplitude (lowest at the peak of testis activity, and highest during the developing phase), and duration of the peaks (shortest when the testes were regressed). In addition, the basal levels changed from being lowest in the regressed phase of the testis cycle, and highest when the gonads were most active. Plasma testosterone concentrations changed in parallel with the cycle of testis size and were correlated with the fluctuating levels of LH. Each episodic peak in plasma LH was associated with an increase in the levels of testosterone, beginning after 0-30 min and rising to a peak at 60-90 min; the speed and magnitude of the response being greatest when the testes were largest, but was not correlated with the magnitude of the LH change. Injections of LH releasing hormone (5 mug) stimulated an increase in plasma LH and testosterone proportional to the endogenous fluctuations in the hormones at the various stages of the seasonal cycle; LH concentrations were raised to supra-physiological levels after the injections, while testosterone concentrations seldom exceeded the normal peak values at any stage. These observations are used to discuss the role of the hypothalamus in the control of male seasonality with emphasis on the dynamic interplay between the hypothalamus, pituitary and testis.

Use of a pulsed infusion of luteinizing hormone releasing hormone to mimic seasonally induced endocrine changes in the ram.

Adult Soay rams were housed indoors under natural lighting during the spring non-mating season when gonadotrophin secretion was low. Four animals received small doses (100 ng or 500 ng) of synthetic LH releasing hormone (LH-RH) infused into the jugular vein by a mechanical device for 60 s every 2 h for 33-57 days: two other rams acted as controls. The prolonged treatment with LH-RH resulted in growth of the testes and the development of the sexual skin flush; these effects were lost when treatment stopped. The plasma concentrations of LH, FSH and testosterone were low at the beginning; each short infusion of LH-RH resulted in a transitory increase in the level of LH and testosterone while the concentration of FSH was only marginally affected. After prolonged treatment with 500 ng pulses of LH-RH the plasma concentrations of all three hormones were permanently raised. The response to the individual injections of LH-RH was also modified, the peak in LH being reduced in amplitude but more prolonged while the FSH and testosterone responses were both enhanced. When the pulsed infusion was stopped the concentration of LH and testosterone declined rapidly while the decline in FSH levels took many days. These endocrine changes induced by the pulsed infusion are comparable to those that occur naturally in the ram during testicular redevelopment before the mating season.

Differential control of luteinizing hormone and follicle-stimulating hormone by luteinizing hormone releasing hormone in the ram.

Adult Soay rams with low concentrations of gonadotrophins in the circulation as a result of 12 weeks of exposure to long daylengths (16 h light : 8 h darkness) were given small doses (100 ng) of synthetic luteinizing hormone releasing hormone (LH-RH) into the jugular vein two, four or seven times/day for 10 days. Each injection of LH-RH induced a transitory increase in the concentration of LH and testosterone in the plasma, whereas the concentration of FSH showed little immediate change. After repeated treatment with pulses of LH-RH, the responses of LH and testosterone became slightly enhanced and the plasma concentration of FSH became permanently raised; these changes were most conspicuous in the animals receiving the most frequent injections. At the end of the study when the injections of LH-RH were stopped, the concentrations of LH and testosterone remained low but the concentrations of FSH continued to be maintained at a high level for at least 24 h.

Central effects of photoperiod on reproduction in the ram revealed by the use of a testosterone clamp.

Over a 3-year period eight adult Soay rams were exposed to an artificial lighting regimen of alternating 16-week periods of long days (16 h light: 8 h darkness; 16L : 8D) and short days (8L : 16D) to induce a seasonal cycle in reproduction and wool growth every 32 weeks. Early in the study the rams were castrated (four during long days and four during short days) and 48 weeks later the castrated animals were each given an s.c. implant of testosterone to increase the blood plasma concentration of testosterone to 14-20 nmol/l. The changes in the concentrations of LH, FSH and testosterone were measured in blood samples collected once or twice weekly while records were made of the changes in the size of the testes (before castration), the intensity of the sexual skin flush, the expression of aggressive and sexual behaviour and the rate of wool growth. The results showed that in the castrated rams there were only minor changes in the blood levels of LH, FSH and the expression of aggressive behaviour related to the 32-week light cycle, while the sexual skin flush was permanently absent. However, after the commencement of the constant testosterone therapy, there were major changes in all the reproductive parameters related to the lighting regimen with a similar temporal relationship as observed in the rams before castration. Cyclic variation in wool growth occurred throughout the study related to the changes in photoperiod but this was not markedly affected by castration and testosterone replacement.(ABSTRACT TRUNCATED AT 250 WORDS)

Endogenous opioids and the control of seasonal LH secretion in Soay rams.

Soay rams were treated with naloxone and/or morphine at different stages of their annual reproductive cycle to study the role of endogenous opioid peptides in the control of pulsatile LH secretion. The responses in intact rams were compared with those shown by pinealectomized (PNX) or superior cervical ganglionectomized (SCGX) rams which had a different annual testicular cycle. Naloxone (4-6 mg/kg i.v.) given to intact rams at four times of the year induced significant increases in LH pulse frequency in the breeding season in September and December, but minimal responses in the non-breeding season in June. Similar treatments given to PNX or SCGX rams induced good responses in March, June and September and the poorest response in December; the different seasonal pattern between the intact and PNX/SCGX rams was correlated with differences in the timing of their testicular cycles. Morphine (1 mg/kg i.v.) induced a significant decrease in LH pulse frequency when given to intact rams in October, but no significant effects were observed when morphine was given to sexually inactive rams in early July. Naloxone (1 mg/kg i.v.) given concurrently with morphine in October reversed the suppressive effect and resulted in an actual increase in LH pulse frequency above pretreatment levels. Morphine-treated rams showed normal LH responses to injections of LH-releasing hormone (LHRH) indicating that the site of opiate inhibition was on hypothalamic LHRH secretion rather than on pituitary LH release. Chronic treatment of intact and PNX rams with naloxone (1 mg/kg every 4 h for 7 days) in April and October produced the expected acute increase in LH pulse frequency in the intact rams in October, and at both times of year in the PNX rams, however there was no sustained increase in LH secretion in response to chronic naloxone in any of the treatment groups. The response to the second naloxone injection was much reduced and was absent after 3 days; responsiveness to naloxone was restored within 2 days of stopping the chronic treatment. The overall results indicate that endogenous opioid mechanism is involved in the tonic inhibition of LH secretion and that this mechanism is most active in the breeding season when both naloxone and morphine have marked effects on pulsatile release of LH. Regulation of endogenous opioids in the hypothalamus may be part of the mechanism by which environmental factors modulate steroid negative-feedback control of LHRH and thus LH secretion in seasonally breeding mammals.