Initiation of the breeding season in ewe lambs and goat kids with melatonin implants

Sixty-eight female and 4 male lambs of the Chios breed born in autumn (September–October) and 48 female and 4 male kids of the Damascus breed also born in autumn (November), were used to evaluate the effect of melatonin implants (Regulin^®) on the initiation of the breeding season. For each species and sex, half of the animals were either left untreated to serve as controls (C) or received ear implants (females one and males two implants) of melatonin (MEL) in May (spring). Each C and MEL group of females was kept separately and away from males for 5 weeks after implantation and then, C and MEL males were joined with the respective groups for 45 (sheep) and 35 days (goats). Animals not conceiving during the early June–July breeding season, were mated again in September (natural breeding season). The reproductive performance and the 60-day milk yield of animals giving birth during the two seasons were compared. Early in the season (mid November–December), significantly more MEL than C ewes and goats gave birth (75.5% versus 23.5%, p < 0.01; 70.8% versus 37.5%, p < 0.05, respectively). For both species, most of the other characteristics examined were significantly affected by season, but not by treatment. Females giving birth early in the season were approximately 80 days younger at parturition and recorded a lower body weight at mating than those giving birth in the natural February–March season. In the first mating period, the animals mated and conceived had similar body weights to those not mated. The natural, compared with the early June–July mating season, was associated in sheep with higher litter weight at birth (7.0 kg versus 5.8 kg, p < 0.05) and in sheep and goats with a higher number (p < 0.05) of offspring born live (1.78 versus 1.38; 2.0 versus 1.61, respectively) and weaned (1.74 versus 1.36; 1.82 versus 1.35, respectively). Total litter size at birth and 60-day milk yield after weaning, were not significantly different between the two seasons. It could be concluded that melatonin implants administered during the last month of spring in autumn-born female Chios sheep and Damascus goats, advanced the initiation of their breeding season by about 80 days when joined with young males also treated with melatonin implants. The overall reproductive performance and milk yield of animals breeding early, was satisfactory and comparable to that of animals breeding in the natural breeding season.     

Effect of melatonin on daily LH secretion in intact and ovariectomized ewes during the breeding season.

This study was conducted to find out whether daily LH secretion in ewes may be modulated by melatonin during the breeding season, when the secretion of both hormones is raised. Patterns of plasma LH were determined in luteal-phase ewes infused intracerebroventricularly (icv.) with Ringer-Locke solution (control) and with melatonin (100 microg/100 microl/h). Response in LH secretion to melatonin was also defined in ovariectomized (OVX) ewes without and after estradiol treatment (OVX+E2). Basal LH concentrations by themselves did not differ significantly before, during and after both control and melatonin infusions in intact, luteal-phase ewes. However, single significant (P<0.05) increases in LH concentration were noted during the early dark phase in the control and 1h after start of infusion in melatonin treated ewes. In both OVX and OVX+E2 ewes, melatonin decreased significantly (P<0.01, P<0.05, respectively) mean plasma LH concentrations as compared to the levels noted before the infusions. In OVX+E2 ewes, a single significant (P<0.05) increase in LH occurred 1h after start of melatonin treatment, similarly as in luteal-phase ewes. No significant differences in the frequencies of LH pulses before, during and after melatonin infusion were found in all treatments groups. In conclusion, melatonin may exert a modulatory effect on daily LH secretion in ewes during the breeding season, stimulating the release of this gonadotropin in the presence of estradiol feedback and inhibiting it during steroid deprivation. Thus, estradiol seems to be positively linked with the action of melatonin on reproductive activity in ewes.     

Importance of changing photoperiod and melatonin secretory pattern in determining the length of the breeding season in the Suffolk ewe

Three groups of ovariectomized Suffolk ewes bearing s.c. Silastic implants of oestradiol were subjected to a 90-day priming treatment of an inhibitory long photoperiod (16 h light/day; 16L:8D). On Day 0 of the experiment, they were moved to stimulatory photoperiods. One control group was transferred to 12L:12D and a second control group was transferred to 8L:16D; both groups remained in those photoperiods to determine the timing of reproductive induction and refractoriness. The experimental group was transferred to 12L:12D on Day 0 and then to 8L:16D on Day 55 to determine whether the further reduction in daylength could delay the development of refractoriness. Reproductive neuroendocrine condition was monitored by serum concentrations of LH and FSH. Both gonadotrophins remained elevated for a longer period of time in the experimental group receiving the second reduction in daylength than in either control group, indicating that the second photoperiodic drop delayed the onset of photorefractoriness. Measurement of 24-h patterns of circulating melatonin suggests that the prolonged stimulation of reproductive neuroendocrine activity in the experimental group resulted from a lengthening of the nocturnal melatonin rise. These findings indicate that refractoriness to an inductive photoperiod can be temporarily overcome by exposure to a shorter daylength, and that the change in duration of the nocturnal increase in melatonin secretion is important in photoperiodic signalling. Thus, in natural conditions, the decreasing autumnal daylength, and the resulting expansion of the nocturnal elevation in melatonin secretion, may be utilized to produce a breeding season of normal duration.     

Nightly duration of pineal melatonin secretion determines the reproductive response to inhibitory day length in the ewe

The pineal controls the reproductive response of ewes to both stimulatory (short) and inhibitory (long) day lengths. Melatonin, a pineal hormone whose nocturnal secretion is entrained by photoperiod, mediates the effect of stimulatory photoperiod. We now report that melatonin also mediates the effect of inhibitory day length, monitored as response to estradiol negative feedback on luteinizing hormone (LH) secretion. Ovariectomized, estradiol-implanted ewes were pinealectomized and intravenously infused with melatonin to restore the nightly melatonin rise. Following transfer from short to long days, and a concurrent switch from short- to long-day melatonin patterns, LH dropped precipitously in pinealectomized ewes, matching the photoinhibitory response of pineal intact controls. LH dropped similarly in pinealectomized ewes when long-day melatonin was infused under short days. Pinealectomized ewes transferred from long to short days displayed a marked LH rise, provided melatonin was also switched to the short-day pattern. LH remained suppressed if long-day melatonin was infused following transfer to short days. These data indicate the nighttime melatonin rise mediates reproductive responses to inhibitory, as well as stimulatory photoperiods; they further suggest the duration of this rise controls suppression of LH under long days. Rather than being strictly pro- or antigonadal, the pineal participates in measuring day length.     

Importance of duration of nocturnal melatonin secretion in determining the reproductive response to inductive photoperiod in the ewe

The pineal gland, through its nocturnal melatonin secretion, mediates the effects of inhibitory (long) and stimulatory (short) photoperiod on reproduction in female sheep. Earlier studies revealed that duration of the nighttime melatonin rise is important in determining the inhibitory effect of day length on reproduction in the ewe. The present study tested whether the duration is also important in mediating the inductive response to short days. Pinealectomized ewes, housed under long days, received a short-day melatonin infusion (16-h duration) for 90 days. Reproductive status was monitored from the response to estradiol negative feedback of luteinizing hormone (LH) secretion. This short-day melatonin pattern led to unambiguous reproductive induction, despite the exposure to inhibitory long days. The increase in serum LH was comparable, in terms of latency and magnitude, to that in pinealectomized controls receiving the same short-day melatonin pattern under short days, and in pineal-intact controls transferred from long to short days. Since the reproductive status conformed to the length of time that melatonin was elevated each day rather than to photoperiod, these results support the conclusion that duration of the nighttime melatonin rise mediates the reproductive response of the ewe to an inductive photoperiod. In all, the melatonin rhythm is considered an integral component of the physiologic mechanism measuring day length; through duration of its nocturnal secretion, melatonin encodes both inhibitory and stimulatory photoperiods.     

Growth hormone and prolactin secretion after hypothalamic deafferentation in pigs

Control of growth hormone (GH) and prolactin (PRL) secretion was investigated in ovariectomized, prepuberal Yorkshire gilts by comparing the effects of anterior (AHD), complete (CHD), and posterior (PHD) hypothalamic deafferentation with sham-operated controls (SOC). Blood samples were collected sequentially via an indwelling jugular catheter at 20-min intervals during surgery and recovery from anesthesia (Day 0) and Days 1 and 2 after cranial surgery. Mean serum concentrations of GH after AHD, CHD, and PHD were reduced (P less than 0.01) when compared with SOC gilts. Furthermore, episodic GH release evident in SOC animals was obliterated after hypothalamic deafferentation. PRL concentrations in peripheral serum of hypothalamic deafferentated gilts remained similar (P greater than 0.05) to those of SOC animals. These results indicate that anterior and posterior hypothalamic neural pathways play a minor role in the control of PRL secretion in the pig in as much as PRL levels remained unchanged after hypothalamic deafferentation. These findings may be interpreted to suggest that the hypothalamus by itself seems able to maintain tonic inhibition of PRL release. In contrast, the maintenance of episodic GH secretion depends upon its neural connections traversing the anterior and posterior aspects of the hypothalamus in the pig.     

Effect of climatic change on the duration of the breeding season in three European thrushes

Capsule Evidence for extension of breeding seasons in Song Thrushes and Blackbirds (multiple‐brooded species) and shortening in Fieldfares (a single‐brooded species).

Aims To analyse breeding data from central Europe during 1964–2006 in relation to climatic conditions operating at breeding and wintering grounds, and to compare breeding phenology in urban and rural habitats.

Methods Using chick ringing data we evaluated long‐term changes in breeding phenology of Fieldfares Turdus pilaris, Song Thrushes T. philomelos and Common Blackbirds T. merula. Changes in phenology were analysed for the 5th percentile, median, 95th percentile, inter‐quantile range (IQR) and brood size.

Results All thrushes showed consistent trends towards earlier onset of breeding. However, there was a contrasting pattern in the later phenological characteristics (median, 95th percentile and IQR) that were associated with single‐ versus multiple‐brooded species. The single‐brooded Fieldfares revealed an advancing trend in all phenological phases, which caused shortening of its breeding period. In contrast, the multiple‐brooded Song Thrushes and Blackbirds delayed both medians and the 95th percentiles of breeding dates leading to extension of their breeding seasons. Temperatures at both wintering and breeding grounds were generally negatively associated with the onset of breeding. Urban birds bred significantly earlier than their rural conspecifics, but brood sizes did not differ.

Conclusion Multiple‐brooded species may respond differently to increasing spring temperatures than single‐brooded species.     

Photoperiodic requirements for timing onset and duration of the breeding season of the ewe: Synchronization of an endogenous rhythm of reproduction

Summary A study was conducted to test the hypothesis that different portions of the annual photoperiodic cycle play different roles in timing the breeding season of the ewe, Ovis aries, an animal in which an endogenous rhythm generates the seasonal reproductive transitions. Adult female sheep were pinealectomized to disrupt transduction of photoperiodic cues at 4 times of the year (summer and winter solstices, vernal and autumnal equinoxes), and the effects on seasonal reproductive neuroendocrine activity were evaluated. Time of pinealectomy greatly influenced the subsequent seasonal reproductive cycle such that the following inferences are possible. Lengthening days between the winter and summer solstices synchronize reproductive onset to the appropriate time of year. The relatively long days around the summer solstice act to suppress reproductive activity and forestall the start of the breeding season until late summer/early autumn. The shortening days between the summer solstice and autumnal equinox maintain a normal intensity and duration of reproductive neuroendocrine induction during the impending breeding season. However, the shortening days between the autumnal equinox and winter solstice (i.e., after breeding season onset) do not appear to play a critical role in maintaining the breeding season of that year, but may provide important cues for timing the breeding season of the following year.Abbreviations LH luteinizing hormone Presented in preliminary form at the 21st Annual Meeting of the Society for the Study of Reproduction, 1988, Biol Reprod 38 (Suppl. 1): 184 (Abstract 408). This work was performed in partial fulfillment of the requirements for the degree of Ph.D. at The University of Michigan (to N.L.W.), and was funded by NIH-HD-18337 and NIH-HD-18258, T-32-HD-07048, the Institut National de la Recherche Agronomique (France) and the Office of the Vice President for Research of The University of Michigan.     

Effect of transport on pulsatile and surge secretion of LH in ewes in the breeding season.

The aim of this study was to elucidate the mechanism(s) involved in stress-induced subfertility by examining the effect of 4 h transport on surge and pulsatile LH secretion in intact ewes and ovariectomized ewes treated with steroids to induce an artificial follicular phase (model ewes). Transport caused a greater delay in the onset of the LH surge in nine intact ewes than it did in ten ovariectomized ewes (intact: 41.0 +/- 0.9 h versus 48.3 +/- 0.8 h, P < 0.02; ovariectomized model: 40.8 +/- 0.6 h versus 42.6 +/- 0.5 h, P < 0.02). Disruption of the hypothalamus-pituitary endocrine balance in intact ewes may have reduced gonadotrophin stimulation of follicular oestradiol production which had an additional effect on the LH surge mechanism. In the ovariectomized model ewes, this effect was masked by the exogenous supply of oestradiol. However, in these model ewes, there was a greater suppression of maximum LH surge concentrations (intact controls: 29 +/- 4 ng ml-1 versus intact transported 22 +/- 5 ng ml-1, P < 0.02; ovariectomized model controls: 35 +/- 7 ng ml-1 versus model transported 15 +/- 2 ng ml-1, P < 0.02). Subsequent exposure to progesterone for 12 days resulted in the resumption of a normal LH profile in the next follicular phase, indicating that acute stress leads to a temporary endocrine lesion. In four intact ewes transported in the mid-follicular phase, there was a suppression of LH pulse amplitude (0.9 +/- 0.3 versus 0.3 +/- 0.02 ng ml-1, P < 0.05) but a statistically significant effect on pulse frequency was not observed (2.0 +/- 0.4 versus 1.7 +/- 0.6 pulses per 2 h). In conclusion, activation of the hypothalamus-pituitary-adrenal axis by transport in the follicular phase of intact ewes interrupts surge secretion of LH, possibly by interference with LH pulsatility and, hence, follicular oestradiol production. This disruption of gonadotrophin secretion will have a major impact on fertility.     

Refractoriness to inhibitory day lengths initiates the breeding season of the Suffolk ewe

Recent evidence indicates that the breeding season of the Suffolk ewe ends because of loss of response to a day length that was previously inductive. This condition of photorefractoriness could potentially also initiate reproduction, as is the case in several long-day breeding rodents. In this study we determined if ewes enter their breeding season because they become refractory to the long ambient photoperiods of late summer. On the summer solstice, 3 groups of ovariectomized ewes (n = 6) bearing s.c. Silastic implants of estradiol (OVX + E) were placed in different day length treatments: 1) natural photoperiod; 2) artificial photoperiod, stimulating natural day lengths; or 3) artificial photoperiod equivalent to that of the summer solstice (16.25L). Entry into the breeding season is associated with a striking (greater than 30-fold) increase in circulating levels of luteinizing hormone (LH). Timing of the onset of the breeding season was not delayed in ewes maintained on the summer solstice photoperiod; LH levels rose simultaneously in all groups. We conclude that ewes normally begin breeding not because they are actively driven to do so by decreasing or short days, but because they become refractory to prevailing long days. Because the pattern of circulating melatonin, which is known to transduce the photoperiodic message, remained entirely appropriate to day length, we believe that the mechanism responsible for photorefractoriness resides in the postpineal processing of the melatonin signal.     

Effect of small doses of naloxone on the pulsatile secretion of prolactin in the crossbreed ewe during the non-breeding season

The objective of this work was to study the effect of small doses of naloxone (Nx) on the pulsatile secretion of prolactin (Pr). For this purpose 12 crossbreed ewes were selected and allocated to three groups of four. Group 1 was treated with two injections (at 7 and 19 h) of 40 microg of GnRH. Group 2 was treated with two i.m. injections (at 7 and 19 h) of 0.5mg of naloxone. And the control group 3 was sham treated with injections of 3 ml saline. Blood samples were taken at 20 min intervals during six consecutive hours after injections. When ewes were treated at 7h no significant changes were observed in concentrations of prolactin following treatment with GnRH. Values fluctuated between 200 and 210 ng/ml. In group 2 treated with naloxone there was no change in plasma Pr concentrations during the first 100 min of sampling, however 60 min after Nx treatment Pr decreased significantly (p<0.01) and thereafter Pr plasma levels were consistently less (p<0.001) than control and GnRH treated ewes for the duration of the experiment. The response of the three groups after the second injection (19 h): After the injection of GnRH plasma Pr levels followed much the same pattern observed after the initial treatment, Pr concentrations were similar to those of control ewes. Ewes treated with a second small dose of naloxone (0.5mg i.m.) however, showed a decrease in plasma Pr 60 min after the administration of the endogenous opioid antagonist. Thereafter Nx treated ewes had lesser (p<0.001) plasma Pr levels until the termination of the experiment. It was concluded that Nx an opioid antagonist administered in small intermittent doses can alter Pr plasma concentrations in the ewe, showing that endogenous opioids are important modulators of endocrine function and that the administration of small intermittent doses of opioid antagonists produce significant endocrine changes in ewes.     

Failure of melatonin to influence endogenous opioid effects on LH secretion in the anoestrous ewe

In May mature seasonally anoestrous ewes were implanted with melatonin which advanced the onset of cycles by about 1 month. The LH response to an opioid antagonist, WIN-3, was determined 5, 15, 25 and 60 days after melatonin implantation, by intravenous administration of WIN-3 (12.5 mg/dose) 4 times at 15-min intervals during both the 1st and the 5th hour of an 8-h treatment period. There was no effect of WIN-3 at 5, 15 and 25 days after melatonin implantation. At 60 days LH concentration and pulse frequency were significantly increased (P less than 0.05 and less than 0.01 respectively) in response to WIN-3 treatment, but only in those animals which had begun reproductive cycles, an effect known to be mediated by the presence of progesterone. We were therefore unable to find evidence to support the hypothesis that the influence of melatonin in advancing the breeding season may be via an opioidergic pathway.     

Effect of feeding melatonin to red deer (Cervus elaphus) on the onset of the breeding season

LHRH-stimulated LH and FSH secretion was studied in hemipituitaries, in vitro, obtained from several dystrophic mouse mutants (male: 129/ReJ-dy; 129B6F1/J-dy; C57BL/6J-dy and C57BL/6J-dy2J; female: 129B6F1/J-dy) and a dystrophic hamster mutant (male and female CHF-147). Without exception, pituitary tissue from dystrophic animals released significantly more FSH than did tissue obtained from controls. LH secretion was more variable; in the male mice released was inhibited, whereas in the male dystrophic hamsters secretion was elevated above normal. The female mouse mutant pituitary released more LH whereas in the female hamster LH secretion was normal. The reduction in body weight of the mutants studied could have contributed to the observations of impaired anterior pituitary function.     

Social cues can play a role in timing onset of the breeding season of the ewe

Female Suffolk sheep were pinealectomized around the vernal equinox to eliminate the major environmental input to the reproductive system (photoperiod) and then either isolated from, or maintained with, pineal-intact gonad-intact sheep. The ewes were ovariectomized and treated with constant-release oestradiol implants and reproductive state was monitored by measuring serum LH concentrations. Pinealectomized ewes that were isolated from the normal flock showed a 2 1/2-month delay in onset of the seasonal rise in LH values compared with that of pineal-intact controls (18 November vs 5 September). On the other hand, pinealectomized ewes that were maintained with the flock showed an onset of the seasonal rise in LH that was not delayed. These results suggest a timekeeping role for social cues for timing onset of the breeding season in an animal that normally relies on photoperiodic signals for temporal regulation of the seasonal reproductive cycle.