Passive immunization against chicken vasoactive intestinal polypeptide suppresses plasma prolactin and crop sac development in incubating ring doves

Passive immunization of incubating ring doves with daily injections of sheep anti-chicken vasoactive intestinal polypeptide (cVIP) serum prevented the proliferation of crop sac tissue observed in control doves given nonimmune serum. Daily injections of anti-cVIP serum did not prevent crop sac development in nonbreeding doves simultaneously treated with ovine prolactin. The concentrations of plasma prolactin were significantly depressed in birds given anti-cVIP serum although this effect became progressively less pronounced during the course of the 7- or 14-day treatment periods. Body weights and weights of regressed reproductive organs were unaffected by treatment with anti-cVIP serum and did not differ significantly from control birds. Doves showing a decreased prolactin response to anti-cVIP serum treatment developed an immune response to sheep serum which may have immunoneutralized the administered antibody. Concentrations of plasma LH were not consistently affected by anti-cVIP serum administration and were low throughout the study. The depression in plasma LH normally seen in females after their young hatch was not observed in females treated with anti-cVIP serum. No effect of treatment was observed upon the birds' incubation behavior or in their readiness to feed and brood their young. These results suggest that in the ring dove, VIP is the physiological prolactin-releasing factor responsible for stimulating prolactin secretion and consequently the development of the crop sac, during incubation. They further indicate that increased concentrations of plasma prolactin may not be essential for gonadal regression or the maintenance of full incubation and brooding behavior in ring doves under laboratory conditions.     

Facilitation of parental behavior in ring doves by systemic or intracranial injections of prolactin

Systemic administration of ovine prolactin (PRL) has been previously reported to stimulate parental feeding behavior toward 7-day-old foster squabs by nonbreeding ring doves with previous breeding experience. The first of the present experiments reexamined this claim in experienced, nonbreeding birds given test squabs of different ages. Each visually isolated male and female dove was given twice-daily subcutaneous injections of ovine PRL or vehicle for 7 days and then tested for parental responses toward a single 1-3, 6-8, or 11- to 13-day-old foster squab. Prolactin significantly increased the incidence or frequency of parental regurgitation-feeding episodes in tests with all three squab age groups and, in addition, increased the incidence of parental feeding invitations (squab-oriented bill openings) in tests with 6- to 8-day-old squabs. A second study explored the degree to which PRL can act directly on the central nervous system to facilitate parental activity in the absence of peripheral cues generated from PRL-induced changes in other target organs, such as those associated with crop sac growth and distension. In this experiment, 6- to 8-day-old test squabs were used to determine if parental behavior is enhanced by twice-daily intracerebroventricular (ICV) injections of PRL in doses below those required to stimulate peripheral target organs. Injection schedules and behavior testing procedures were the same as those used in Experiment 1. However, half of the ICV PRL-treated and ICV vehicle-treated birds were food deprived for 16 hr before and during the test in order to control for PRL-induced hyperphagia and resulting crop sac distension, which could confound the results by generating peripheral stimuli conductive to the display of regurgitation-feeding behavior. Intracranial injections of prolactin significantly increased the incidence of feeding behavior, parental feeding invitations, and crouching or sitting in the nest in food-deprived doves but not in freely fed animals. Empty crop sac weights of freely fed and food-deprived PRL-treated birds were not increased above control values, thus indicating that ICV PRL treatment did not result in significant stimulation of peripheral target organs. These results demonstrate a facilitative action of PRL on regurgitation-feeding responses and associated parental behaviors that is not restricted to squabs of one particular age range. They also indicate that PRL is capable of acting directly on the brain to promote these activities in the absence of PRL-induced changes in the crop sac and other peripheral target organs.     

The Roles of Stimuli from Young, Previous Breeding Experience, and Prolactin in Regulating Parental Behavior in Ring Doves (Streptopelia risoria)

In addition to stimulating crop "milk" formation in ring doves, prolactin (PRL) may promote the parental regurgitation behavior that transfers this "milk" to the young at the time of hatching. Although earlier studies suggest that previous breeding experience is an important modulator of PRL-induced parental regurgitation behavior in ring doves, the ways in which experience, hormones, and stimuli from young interact to promote parental behavior have not been well characterized in this species. In the first study, untreated, nonbreeding female doves with and without previous breeding experience were given 10 daily parental behavior tests (2 h/day) with a hungry 5- to 10-day-old foster squab. Experienced females exhibited a higher incidence of regurgitation behavior, defensive behavior, and crouching or sitting in the nest than did inexperienced females. In a second study, nonbreeding females were given 10 daily tests for parental behavior while they received sc injections of ovine PRL or vehicle. Prolactin reduced squab-directed aggression and increased the incidence of regurgitation feeding behavior of foster squabs in both experienced and inexperienced females. However, the average number of regurgitation feeding acts displayed by those PRL-treated females that showed the behavior was over eight times higher in experienced females than in inexperienced females. Previous experience also enhanced the stimulatory effects of PRL on defensive behavior and crouching or sitting in the nest. The parental behavior exhibited by nonbreeding, PRL-treated experienced females was qualitatively and quantitatively similar to that observed in normally breeding females during a single test with their own hungry 5- to 10-day-old squabs. These findings indicate that PRL and previous breeding experience both enhance the parental responsiveness of nonbreeding female doves and that under optimal hormonal, experiential, and squab exposure conditions, nonbreeding doves exhibit levels of parental activity that rival those of normally breeding parents.     

Effects of presence of squabs upon plasma concentrations of prolactin and LH and length of time of incubation in ringdoves on "extended" incubatory patterns

The effects of a brief exposure to newly hatched squabs upon the concentrations of plasma prolactin and LH secretion and length of incubation time in ring doves were investigated using a novel experimental procedure. After 5 days of incubation birds were deprived of their nests for 3 days. On return to their nests, the birds immediately resumed incubation and laid a second clutch of eggs after 5 days at which time the first clutch was removed. A consequence of this procedure was that at the time of hatching, concentrations of plasma prolactin had passed their peaks and were falling to baseline values, while concentrations of plasma LH were beginning to increase. This experimental manipulation made it possible to demonstrate that brief exposure to newly hatched squabs significantly affects the concentration of plasma prolactin, an effect which was more pronounced in females than in males. This increased secretion of prolactin was associated with a reduction of plasma LH which reached significance in females but not in males. Birds exposed to squabs spent a significantly longer period of time on their nests as compared to those birds not exposed to squabs. This observation is consistent with a role for prolactin in maintaining readiness to incubate.     

Concentrations of plasma prolactin and luteinizing hormone following nest deprivation and renesting in ring doves (Streptopelia risoria)

Ring doves with increased plasma prolactin and low plasma LH (Group A) or with low plasma prolactin and low plasma LH (Group B) which had been incubating sterile eggs for 12 or 18 days, respectively, had their nests and eggs removed for 3 days. Upon nest return, observations were made on the birds' readiness to renest and on changes in plasma prolactin and LH. Birds from Group A demonstrated a far greater tendency to resume incubation than birds from Group B. Nest deprivation resulted in a sharp fall in the concentration of plasma prolactin in birds which were deprived of their nests after 12 days of incubation (Group A). Following resumption of incubation no subsequent increase in the prolactin levels was observed in Group A or B. The concentration of plasma LH rose sharply after nest deprivation in both sexes of both groups and declined after return of the nests. Birds in Groups A and B which returned to their nests laid a new clutch of eggs while continuing to incubate. The total length of uninterrupted sitting following nest return was 20.9 +/- 0.48 days (n = 8). These results suggest that (1) once the mechanism responsible for the increase in plasma prolactin during incubation is disrupted, it cannot be reactivated unless the whole reproductive cycle is repeated. (2) The inhibition of LH secretion during incubation involves neural mechanisms which do not necessarily involve the anti-gonadotrophic action of prolactin.     

Hypothalamic monoamine turnover in ring doves (streptopelia risoria), courting,incubating and brooding young

1. The turnovers of hypothalamic 5-hydroxytryptamine (5HT), dopamine (DA) and noradrenaline (NE) were measured in male and female ring doves (Streptopelia risoria) at three stages of the breeding cycle: courtship, 3 days after pairing; early incubation, 1–2 days after egg laying; and brooding, 1–3 days after the squabs had hatched.2. In both sexes plasma LH decreased progressively from courtship through incubation to brooding young. Crop sacs were fully developed in doves brooding young but not at other stages of the reproductive cycle, indicating increased concentrations of plasma prolactin.3. The turnovers of 5HT and DA in both sexes were significantly higher (P<0.001) in doves brooding young than in birds incubating eggs or nest building. The turnover of DA was higher in females than in males at the onset of incubation. The turnover of NE was lower (P > 0.01) in females at the onset of incubation than during courtship or brooding.4. Increased turnover of hypothalamic DA may be more closely related to brooding behaviour than to changes in prolactin or LH secretion.5. Increased hypothalamic 5HT turnover in brooding doves appears to be more directly related to crop sac development, and by inference increased prolactin secretion, than to depressed plasma LH concentrations.     

Gonadal hormones determine sex differences in timing of incubation by doves.

Male and female ring doves express a sexually dimorphic pattern of incubation. The dimorphism is temporal rather than motoric. The male incubates for a block of time in the middle of the day and the female incubates the rest of the time. The present study explored the role of gonadal hormones in the control of the temporal dimorphism. Female-female pairs incubated their eggs, but it could not be predicted which of the partners would be sitting on the nest at any given time. Male-male pairs did not incubate and instead destroyed the nests that were provided and displayed aggressive behavior. Some intact males incubated when paired with gonadectomized males, although the castrates tended to ignore the nest. In contrast, when castrated birds were given heterotypical hormonal replacement therapy, they and their same-sex partners incubated the eggs that had been provided, with the gonadectomized birds sitting at a time appropriate to the hormonal state. The results indicate that gonadal hormones influence not only the expression of incubation behavior, but also its phase and duration.     

Termination of incubation in doves: Influence of egg fertility and absence of mate

The present experiments examine the role of the eggs, the mate, and hormonal state in the termination of incubation by ring doves. Doves given either infertile eggs or newly laid fertile eggs at midincubation did not prolong incubation significantly beyond that observed in doves with hatching eggs. However, there was substantial lack of synchrony between mates in the experimental groups. Half the experimental females laid new eggs, often without interrupting incubation. To clarify the role of the mate in maintaining sitting, pairs were separated on Days 1, 5, or 10 of incubation. Initially birds in each group incubated alone throughout the day. The decline in sitting was more rapid in pairs which had been separated early in incubation. To explore the hormonal correlates of the termination of incubation, mates were separated on the 10th day of incubation and observed until they quit sitting. At this time, males showed substantial crop development. Females had little crop development and large preovulatory folliclels, indicating readiness to lay new eggs despite the absence of the male. We conclude that the termination of incubation by parent doves is delayed by the presence of the mate and this, in turn, prevents follicular development and recycling by the female. Cues from the offspring are important in synchronizing the behavior of the parents and also in preventing a new breeding cycle.     

Hormonal Induction of Male Courtship Behavior in the Japanese Newt, Cynops pyrrhogaster

In the breeding season, the sexually mature male newt, Cynops pyrrhogaster, vibrates the tail in front of the female at an early stage of courtship. Effects of prolactin (PRL), gonadotropin (GTH), and sex steroids on this behavior were investigated in the male paired with the female receiving PRL and GTH. The behavior was elicited in the sexually inert male by injections of PRL of bovine, ovine, or bullfrog origin and human chorionic gonadotropin or bullfrog LH and FSH in combination. The effect of PRL or GTH alone was less marked than that of PRL plus GTH, especially in terms of frequency of the behavior. In the hypophysectomized male, combination of PRL and GTH significantly increased both the incidence and frequency of the behavior. However, PRL alone was not effective, and the effect of GTH alone was less pronounced than that in the intact animal receiving GTH injections. The effect of GTH was nullified by castration. In the PRL-treated castrated animal, testosterone or dihydrotestosterone, but not estradiol, was effective in inducing the behavior.     

Contributions of reproductive experience to observation-maintained crop growth and incubation in male and female ring doves

Previous work has established that experienced male ring doves (Streptopelia risoria) can maintain prolactin-dependent crop growth and readiness to incubate by observing an incubating partner. We report that this is also true for female ring doves. The role of experience in this phenomenon was examined in separate experiments with males and females. Observation of an incubating mate from 3 days after completion of egg laying is sufficient to maintain crop growth and incubation in both male and female ring doves in their second, but not in their first, reproductive cycle. Male and female doves in their first cycle must incubate for a greater part of the cycle before observation of an incubating mate is an effective stimulus; there are no differences between first and second cycle doves separated by a glass plate from the mate and nest 8 days after laying. Experience obtained within the first cycle apparently ensures that previously neutral stimuli come to elicit prolactin secretion. The effectiveness of these stimuli is reinstated early in a second cycle.     

Vasoactive intestinal polypeptide-like immunoreactivity during reproduction in doves: influence of experience and number of offspring

To assess the possibility that vasoactive intestinal polypeptide (VIP) plays a role in naturally occurring changes in prolactin secretion in ring doves, we used immunohistochemical techniques to measure VIP-like immunoreactivity in the brain as a function of stage of the reproductive cycle. Differences between parental and nonparental birds in VIP profiles were detected in the ventral portion of the infundibular region. More specifically, there is an increase in cell size and staining intensity in the ventral infundibulum of breeding birds compared to simultaneously processed tissue taken from control animals. In both sexes, an increase in size of VIP-like immunoreactive cells is detectable during courtship and early incubation, anticipating the increase in plasma prolactin levels. VIP cell size remains elevated from about Incubation Day 14 to Brooding Day 14, and a steady decrease is observed during the remaining posthatching period, as squab begin to feed independently. Compared to parents rearing one squab, those with two young have a prolonged interval of increased infundibular VIP immunoreactivity. Furthermore, doves with no previous experience of a breeding cycle exhibit prolonged VIP-like immunoreactivity compared to experienced parents, paralleling previously described differences between these groups in parental behavior.     

The transition from sitting on eggs to sitting on young in ring doves, Streptopelia risoria: squab-egg preferences during the normal cycle.

Ring doves of both sexes sit on young squabs after hatching in much the same manner as they sit on eggs before hatching, but this study demonstrates that the preferred stimulus varies with the state of the animal. A simultaneous, squab-egg choice test was given on days 1, 4, 10, and 13 of incubation and on the day following hatching in normal reproductive cycles of experienced and naïve male and female ring doves. Naïve doves were more likely than experienced doves to choose eggs throughout the cycle (P<0·005) and, overall, eggs were more likely to be chosen during early incubation and squabs posthatching (P<0·005).     

Gonadal and hormonal characteristics of captive adult mourning doves during the nesting cycle

Paired mourning doves ($\text{Zenaida}$$\text{macroura}$) of a captive breeding colony were selected and sacrificed on specific day intervals postlaying and posthatching during 1976–1977. Blood plasma samples were analyzed for androgens, estradiol, and progestins using radioimmunoassays. Testes, ovary and oviduct weights, spermatozoan numbers, diameter of the three largest follicles in the ovary, the presence or absence of eggs in the reproductive tract, and crop gland characteristics also were recorded. Differences (P<0.05) existed between nonbreeding and breeding pairs in testes weights, spermatozoan numbers, and ovary and oviduct weights. No distinct cyclic pattern was observed in gonadal variables among breeding males and females during the nesting cycle, although trends were observed. No differences (P > 0.05) in androgen and progestin concentrations in males, or estradiol and progestin concentrations in females, between nonbreeding and breeding pairs, or among breeding pairs during different stages of the nesting cycle, were observed. Analysis of reproductive variables by stage of crop gland activity tended to indicate an inverse relationship between gonadal and crop gland activity in both male and female doves during the nesting cycle.     

Prolactin-Brain Interactions and Reproductive Function

SYNOPSIS. There is mounting evidence that prolactin influencesreproductive function in several vertebrate species via directaction on the central nervous system. In most instances, however,the basic properties of these prolactin-induced alterationsin brain function, the neurochemical mechanisms underlying theseeffects, and the identity of the prolactinlike molecules thatmight normally promote these changes have yet to be adequatelycharacterized. Several properties of the prolactin-brain relationshipthat have emerged from our work in the ring dove (Streptopeliarisoria) are described in this paper and discussed in relationto similar work in other species. Receptor binding studies usingradiolabelled mammalian prolactin preparations consistentlyindicate the existence of saturable, high affinity binding sitesfor prolactin in the brains of several vertebrate species. Theapparent concentration of these sites in the diencephalon iscorroborated by recent mapping studies in rabbits, ring doves,and three song birds in which in vitro autoradiography and densitometrywas employed. Nevertheless, direct comparisons among the threesongbird species suggest some differences in the precise distributionand/or concentration of prolactin binding sites within the preoptic-hypothalamiccontinuumthat may relate to species differences in the display of prolactin-related incubation behavior and parental activities. Althoughdefinitive evidence is currently lacking, it would appear thatblood-borne prolactin gains access to binding sites in the centralnervous system and that a receptor-mediated, blood-to-cerebrospinalfluid transport process in the choroid plexus may be a majorconduit for such uptake. Recent findings from several vertebratespecies also suggest that the brain may synthesize one or moreprolactin-like molecules that could conceivably interact withthese binding sites. Direct actions of prolactin and relatedhormones on brain mechanisms underlying reproductive processesare suggested by the profound dose-dependent suppression ofgonadotropin secretion and gonadal activity in ring doves byintracerebroventricular administration of prolactin or growthhormone at doses that are below those required for significantperipheral actions. A major challenge for future investigationwill be to identify the brain site(s) at which these and othereffects of prolactin are exerted and to determine how blood-borneprolactin of pituitary origin, prolactin-like molecules of brainorigin, and growth hormone-like molecules interact with eachother and with other neurochemical systems to promote thesetypes of changes under normal physiological conditions.     

The role of prolactin in reproductive failure associated with heat stress in the domestic turkey

Reproductive failure associated with heat stress is a well-known phenomenon in avian species. Increased prolactin (PRL) levels in response to heat stress have been suggested as a mechanism involved in this reproductive malfunction. To test this hypothesis, laying female turkeys were subjected to 40 degrees C for 12 h during the photo-phase daily or maintained at 24-26 degrees C. Birds in each group received oral treatment with parachlorophenyalanine (PCPA; 50 mg/kg BW/day for 3 days), an inhibitor of serotonin (5-HT) biosynthesis, or immunized against vasoactive intestinal peptide (VIP). Both treatments are known to reduce circulating PRL levels. Nontreated birds were included as controls. In the control group, high ambient temperature terminated egg laying, induced ovarian regression, reduced plasma luteinizing hormone (LH) and ovarian steroids (progesterone, testosterone, estradiol) levels, and increased plasma PRL levels and the incidence of incubation behavior. Pretreatment with PCPA reduced (P < 0.05) heat stress-induced decline in egg production, increase in PRL levels, and expression of incubation behavior. Plasma LH and ovarian steroid levels of heat stressed birds were restored to that of controls by PCPA treatment. As in PCPA-treated birds, VIP immunoneutralization of heat-stressed turkeys reduced (P < 0.05) circulating PRL levels and prevented the expression of incubation behavior. But it did not restore the decline in LH, ovarian steroids, and egg production (P > 0.05). The present findings indicate that the detrimental effect of high temperature on reproductive performance may not be related to the elevated PRL levels in heat-stressed birds but to mechanism(s) that involve 5-HT neurotransmission and the induction of hyperthermia.     

Individual variation in baseline and stress-induced corticosterone and prolactin levels predicts parental effort by nesting mourning doves

Endocrine systems have an important mechanistic role in structuring life-history trade-offs. During breeding, individual variation in prolactin (PRL) and corticosterone (CORT) levels affects behavioral and physiological processes that drive trade-offs between reproduction and self-maintenance. We examined patterns in baseline (BL) and stress induced (SI; level following a standard capture-restraint protocol) levels of PRL and CORT for breeding mourning doves (Zenaida macroura). We determined whether the relationship of adult condition and parental effort to hormone levels in wild birds was consistent with life-history predictions. Both BL PRL and BL CORT level in adults were positively related to nestling weight at early nestling ages, consistent with the prediction of a positive relationship of hormone levels to current parental effort of adults and associated increased energy demand. Results are consistent with the two hormones acting together at baseline levels to limit negative effects of CORT on reproduction while maintaining beneficial effects such as increased foraging for nestling feeding. Our data did not support predictions that SI responses would vary in response to nestling or adult condition. The magnitude of CORT response in the parents to our capture-restraint protocol was negatively correlated with subsequent parental effort. Average nestling weights for adults with the highest SI CORT response were on average 10–15% lighter than expected for their age in follow-up visits after the stress event. Our results demonstrated a relationship between individual hormone levels and within population variation in parental effort and suggested that hormonal control plays an important role in structuring reproductive decisions for mourning doves.     

Prolactin involvement in the regulation of the hypothalamic-pituitary-ovarian axis during the early luteal phase of the porcine estrous cycle.

Our previous in vivo and in vitro studies revealed that prolactin (PRL) affected luteal function during the first days of the porcine estrous cycle. Since the mechanism by which the luteotrophic action of PRL might be mediated was not elucidated, the goal of the present study is to investigate the effects of short term, in vivo administration of PRL on in vitro functions of hypothalamic explants, adenohypophyseal cells and luteal cells of sows. Injections of PRL or saline (performed every 2h) started shortly after the preovulatory LH surge and lasted for 2 or 3 days. Peripheral blood plasma for determination of LH, PRL and progesterone (P(4)) was sampled at 4h intervals. Ovaries, pituitaries and the stalk median eminence (SME) dissected after slaughter were used for in vitro studies. Luteal and adenohypophysial cells as well as hypothalamic tissue were incubated/cultured with different treatments. Medium and plasma levels of GnRH, LH and P(4) were quantified by radioimmunoassays (RIAs). Corpora lutea (CL) were used for LH/human chorionic gonadotrophin (hCG) receptor analysis. In vivo and in vitro treatment with PRL increased the in vitro GnRH release by hypothalamic explants (P<0.05). GnRH-stimulated LH production was enhanced in PRL-treated sows compared to that of control sows (P<0.05). PRL injections had no effect on plasma P(4) concentrations during the treatment period. However, luteal secretion of P(4) (P=0.06) and LH/hCG receptor concentration (P=0.079) tended to be higher in PRL-treated sows in comparison to those of controls. The results indicate that PRL may be involved in the regulation of the hypothalamic-pituitary-ovarian axis at the beginning of the luteal phase of the porcine estrous cycle.     

Melatonin: Failure of pharmacological doses to induce testicular atrophy in the male golden hamster

Effects of graded doses of melatonin on reproductive function of the male golden hamster were studied. Daily injections of melatonin at 17:00 h brought about complete testicular regression when the hamster received 10, 25, and 100 ug melatonin; larger doses of 1,000 ug or 2,500 ug had a partial or no effect on testicular or accessory sex organ weights. The dose response relationship of testicular and accessory sex organ weights to melatonin treatment somewhat resembled an inverted bell-shaped curve. Plasma, LH and prolactin concentrations followed the pattern of the testicular and accessory sex organ weights; small doses of melatonin suppressed plasma, LH and prolactin while larger doses had no suppressive effects. These results indicate that melatonin can exert a reproductive inhibitory effect if injected with small, but not with larger doses.     

Prolactin-induced parental hyperphagia in ring doves: are glucocorticoids involved?

Hyperphagia is a prominent component of the parental behavior repertoire in male and female ring doves and is necessary in order for parents to successfully provision their growing young. Although previous studies implicate both prolactin and the endogenous glucocorticoid, corticosterone, in parental hyperphagia, the functional interactions between these two hormones in regulating changes in feeding activity have not been characterized. These studies examined the possibility that prolactin's orexigenic effects are mediated through the increased secretion of corticosterone. Twice-daily intracerebroventricular (icv) injection of prolactin increased plasma corticosterone concentration in non-breeding doves of both sexes, with males exhibiting more pronounced effects than females. To further test the importance of glucocorticoid signaling in prolactin-induced feeding responses, changes in food intake were investigated in icv prolactin-treated, non-breeding doves following icv infusion of the glucocorticoid receptor antagonist RU38486 or propylene glycol vehicle. No attenuation of prolactin-induced hyperphagia was observed in either sex following co-administration of RU38486 at a dose shown previously to block dexamethasone-induced feeding in doves. These findings suggest that elevated corticosterone titers in blood may contribute to the hyperphagia observed in response to prolactin, but corticosterone signaling through a mammalian-type glucocorticoid receptor is not essential.     

Reproductive effects of 6-chloromelatonin implants and/or injections in male and female Syrian hamsters (Mesocricetus auratus)

Adult female hamsters were injected each afternoon for 9 weeks with 2.5, 15 or 25 micrograms of melatonin or 6-chloromelatonin (CM). Each drug resulted in a significant dose-related depression in uterine, ovarian and anterior pituitary gland weights. Additionally, plasma and pituitary concentrations of prolactin fell with increasing dose of either indole whereas pituitary levels of LH and FSH were positively correlated with dose. There was no difference in effectiveness between the two drugs. Adult male hamsters treated for 10 weeks with daily afternoon injections of melatonin and a blank beeswax pellet had depressed testicular and accessory organ weights and plasma and pituitary concentrations of prolactin. Implantation of a 1 mg melatonin or CM beeswax pellet in hamsters concurrently receiving daily afternoon injections of melatonin prevented the organ weight and hormonal changes, except for plasma prolactin. Adult male hamsters treated for 10 weeks with daily afternoon injections of CM and a blank beeswax pellet had depressed reproductive organ weights and pituitary and plasma concentrations of prolactin; this depression in hormonal values and organ weights was totally prevented if the CM-injected hamsters were also bearing a beeswax-melatonin pellet. In conclusion, 6-chloromelatonin is as effective as melatonin with regards to antigonadotrophic and counter-antigonadotrophic effects in male and female Syrian hamsters.