Courtship interactions stimulate rapid changes in GnRH synthesis in male ring doves

Many birds and mammals show changes in the hypothalamo-pituitary-gonadal (HPG) axis in response to social or sexual interactions between breeding partners. While alterations in GnRH neuronal activity play an important role in stimulating these changes, it remains unclear if acute behaviorally-induced alterations in GnRH release are accompanied by parallel changes in GnRH synthesis. To investigate this relationship, we examined changes in the activity of GnRH neurons in the brains of male ring doves following brief periods of courtship interactions with females. Such interactions have been previously shown to increase plasma LH in courting male doves at 24 h, but not at 1 h, after pairing with females. In the first study, males allowed to court females for 2 h had 60% more cells that showed immunocytochemical labeling for GnRH-I in the preoptic area (POA) of the hypothalamus than did control males that remained isolated from females. To determine whether an increase in GnRH gene expression preceded this increase in GnRH immunoreactivity in the POA, changes in the number of cells with detectable GnRH-I mRNA in the POA were measured by in situ hybridization following a 1 h period of courtship interactions with females. In this second study, courting males exhibited 40% more cells with GnRH-I in this region than did isolated control males. GnRH-immunoreactive neurons in two other diencephalic regions failed to show these courtship-induced changes. Plasma LH was not elevated after 1 or 2 h of courtship. These results demonstrate that the release of GnRH-I in the POA that is presumably responsible for courtship-induced pituitary and gonadal activation is accompanied by a rapid increase in GnRH synthesis that occurs before plasma LH levels increase. We suggest that this increase in GnRH synthesis is necessary to support the extended period of HPG axis activation that is seen in this species during the 5–10 day period of courtship and nest building activity.     

Effects of the antiandrogen cyproterone acetate on reproduction in male and female ring doves

The present experiment was designed to study the role of increased plasma androgens which are seen in male and female doves during courtship. In pairs of doves either the male or female was injected daily with oil or with 6 or 15 mg of cyproterone acetate (CA), starting 3 days before pairing. Cyproterone acetate at both dose levels reduced the amount of bow-cooing shown by the male on the day of pairing, and at the higher dose resulted in a reduction of nest soliciting by injected males and their untreated mates over the 7 days of testing. At the high dose only, the size of the testes was reduced. In pairs where females received CA, there was a decrease in levels of nest soliciting by both partners and ovarian development was blocked. The results are discussed in terms of the possible significance of fluctuating androgen levels found in doves during the prelaying period.     

The effects of photoperiod on androgen-induced reproductive behavior in male ring doves, Streptopelia risoria

The effect of photoperiod on reproductive behavior in male ring doves (Streptopelia risoria) was studied in androgen-injected castrates maintained under long (16L:8D) or short (86:16D) photoperiods. Behavioral recordings were made over a 2-week period during which each male was paired with a female for 6.5 hr/day. There was some indication that males held on long days display higher levels of courtship activity during the initial period following pairing, but the evidence was not conclusive. Day length had no effect upon copulatory behavior. Long-day males exhibited consistently higher levels of nest building than short-day males, indicating that photoperiod affects building through some mechanism other than changes in endogenous androgen levels.     

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.     

Effects of septal lesions on the courtship behavior of male ring doves (Streptopelia risoria)

Male ring doves exhibit several androgen-dependent behavior patterns. Preliminary studies indicated that one of these, the nest-soliciting display, was selectively increased following septal lesions. In the present study castrated male ring doves received a septal lesion followed by daily intramuscular injections of 30 μg testosterone propionate (TP). Although one of the androgen-dependent displays, the bow-coo, was unaffected by the lesions, nest soliciting was significantly elevated during the period of hormone treatment. The restoration of nest-soliciting behavior in the males with septal lesions was even more rapid than that of unlesioned males receiving 200 μg TP/day, although after 12 days of treatment the performances of the latter came to approximate those of the males bearing septal lesions. Males that had received lesions in brain areas other than the septum were no more responsive to 30 μg of TP than were nonlesioned males receiving similar hormone treatment. It is suggested that the influence of the septum differs among the various androgen-dependent displays.     

Glucocorticoids and parental hyperphagia in ring doves (Streptopelia risoria)

These studies explored the possibility that glucocorticoids promote parental care in ring doves by mediating, at least in part, the pronounced increase in food consumption that parent doves exhibit while provisioning their young. Plasma concentrations of the endogenous glucocorticoid corticosterone were found to be significantly higher in breeding females during the posthatching phase than during the incubation period. These differences were not observed in male breeding partners, but sex differences in daily activity rhythms are well documented in breeding doves, and blood sampling at different times of day would be required to adequately characterize the pattern of corticosterone in males during these breeding stages. In studies on nonbreeding doves, twice-daily intracerebroventricular (icv) injections of the synthetic glucocorticoid dexamethasone (DEX) increased food intake by 25-50% in both sexes, and further studies in males revealed that the increase was directly related to the dose of DEX administered. The highest dose of DEX given icv (1.0 microg/day) was not effective in stimulating feeding when given systemically, thereby suggesting that the hyperphagic action of DEX is exerted directly on the central nervous system. The icv infusion of the selective glucocorticoid receptor antagonist RU38486 blocked the hyperphagic effects of twice-daily icv injections of DEX in both sexes. Collectively, these data support the hypothesis that corticosterone contributes to the parental hyperphagia exhibited by breeding doves during the posthatching period. They also suggest that these orexigenic effects are mediated in part by CNS binding sites that resemble mammalian glucocorticoid receptors.     

Neural factors that stimulate ecdysteroid synthesis by the larval ring gland ofDrosophila melanogaster

Summary The larval ring gland ofDrosophila melanogaster is the source of ecdysteroids responsible for larval-larval and larval-pupal molting. An extract prepared from theDrosophila larval central nervous system, that presumably contains prothoracicotropic hormone, elicits a significant and dose-dependent in vitro increase in ecdysteroid synthesis by ring glands from wandering third instar larvae. The synthesis of all three ecdysteroids previously identified as ring gland products is elevated by more than two-fold in the presence of neural extract. The maximum response occurs within 30 min and can be sustained for at least two hours after a 30 min exposure to neural extract. No non-neural tissue extracts evoke a response and most of the prothoracicotropic activity originates in the ventral ganglion. However, while extract prepared from larval brains elicits only a slight increase in ecdysteroid synthesis, it enhances the activity of a submaximal dose of ventral ganglion extract. This suggests that two or more neural factors, at least one from the brain lobes and another from the ventral ganglion, interact to stimulate ecdysteroid synthesis by the larval ring gland.Abbreviations CNS central nervous system - HPLC high performance liquid chromatography - PTTH prothoracicotropic hormone - RIA radioimmunoassay     

Dynamic changes in social dominance and mPOA GnRH expression in male mice following social opportunity

Social competence - the ability of animals to dynamically adjust their social behavior dependent on the current social context – is fundamental to the successful establishment and maintenance of social relationships in group-living species. The social opportunity paradigm, where animals rapidly ascend a social hierarchy following the removal of more dominant individuals, is a well-established approach for studying the neural and neuroendocrine mechanisms underlying socially competent behavior. In the current study, we demonstrate that this paradigm can be successfully adapted for studying socially competent behavior in laboratory mice. Replicating our previous reports, we show that male laboratory mice housed in a semi-natural environment form stable linear social hierarchies. Novel to the current study, we find that subdominant male mice immediately respond to the removal of the alpha male from a hierarchy by initiating a dramatic increase in aggressive behavior towards more subordinate individuals. Consequently, subdominants assume the role of the alpha male. Analysis of brain gene expression in individuals 1 h following social ascent indicates elevated gonadotropin-releasing hormone (GnRH) mRNA levels in the medial preoptic area (mPOA) of the hypothalamus compared to individuals that do not experience a social opportunity. Moreover, hormonal analyses indicate that subdominant individuals have increased circulating plasma testosterone levels compared to subordinate individuals. Our findings demonstrate that male mice are able to dynamically and rapidly adjust both behavior and neuroendocrine function in response to changes in social context. Further, we establish the social opportunity paradigm as an ethologically relevant approach for studying social competence and behavioral plasticity in mammals.     

Insulin and growth factors stimulate rapid posttranslational changes in glucose transport in ovarian granulosa cells

The glucose analogues, 3-O-methyl-D-glucose and 2-deoxy-D-glucose, have been used to characterize glucose transport and its regulation by serum and growth factors in monolayer cultures of granulosa cells obtained from bovine ovaries. Uptake of 3-O-methylglucose was shown to be independent of the Na+-gradient, independent of energy, did not show accelerated exchange, and was stereospecific. Serum withdrawal resulted in a biphasic decrease in initial rates of glucose uptake with half-times for the two phases of 50 minutes and 3 hours. Insulin could prevent the decrease in uptake rates with a half-maximum concentration of 10.0 1/8 3 nM. Insulin was shown to stimulate DNA synthesis with a concentration of half-maximum response of 28 nM. Insulin or serum stimulation of 3-O-methylglucose uptakes in serum-starved cells resulted in a two threefold increase in initial rates, with a time for half-maximum stimulation of 3 minutes. The insulin-stimulated increase was insensitive to cycloheximide and cyanide during the first 30 minutes, and this early, rapid stimulation was also produced by brain FGF (fibroblast growth factor), pituitary FGF, epidermal growth factor, calf serum, and some but not all samples of follicular fluid. Insulin also stimulated 2-deoxyglucose and a-aminoisobutyric acid uptake during the first 5 minutes of addition and these early stimulations were shown to be posttranslational changes.     

Agonistic interactions elicit rapid changes in brain nonapeptide levels in zebrafish

The teleost fish nonapeptides, arginine vasotocin (AVT) and isotocin (IT), have been implicated in the regulation of social behavior. These peptides are expected to be involved in acute and transient changes in social context, in order to be efficient in modulating the expression of social behavior according to changes in the social environment. Here we tested the hypothesis that short-term social interactions are related to changes in the level of both nonapeptides across different brain regions. For this purpose we exposed male zebrafish to two types of social interactions: (1) real opponent interactions, from which a Winner and a Loser emerged; and (2) mirror-elicited interactions, that produced individuals that did not experience a change in social status despite expressing similar levels of aggressive behavior to those of participants in real-opponent fights. Non-interacting individuals were used as a reference group. Each social phenotype (i.e. Winners, Losers, Mirror-fighters) presented a specific brain profile of nonapeptides when compared to the reference group. Moreover, the comparison between the different social phenotypes allowed to address the specific aspects of the interaction (e.g. assessment of opponent aggressive behavior vs. self-assessment of expressed aggressive behavior) that are linked with neuropeptide responses. Overall, agonistic interactions seem to be more associated with the changes in brain AVT than IT, which highlights the preferential role of AVT in the regulation of aggressive behavior already described for other species.     

Vernal changes in the behavioral and endocrine responses to GnRH application in male European ground squirrels

This field study was aimed at examining hypothalamic involvement in the behavioral changes of male European ground squirrels (Spermophilus citellus) before, during, and after the mating season. The effects of exogenous gonadotropin-releasing hormone (GnRH) application on androgen secretion and behavioral patterns were investigated. Animals were captured, bled, and injected intramuscularly with 40 ng/100 g of GnRH. A second plasma sample was collected 40 min after the treatment to document changes in testosterone secretion. Behavioral parameters such as intra-sexual aggression, scent marking, and home range size were compared on the days before and after the stimulation. In the first two phases, before female emergence and during mating, GnRH-injection caused increases in plasma testosterone. In the post-mating phase, initial plasma testosterone levels had decreased and no elevation could be induced. Sham treatment of controls had no effect in any phase. Conditional parameters like emergence body mass and testicular size covaried with androgen increases only in the pre-mating period. Behavioral changes after GnRH administration occurred during the pre-mating period. Intra-sexual aggression, scent marking, and home range size increased significantly in experimental individuals. Later, during mating and post-mating, we found no behavioral changes associated with the GnRH treatment or the testosterone increase. The results demonstrate changes in the endocrine and behavioral sensitivity to GnRH application, according to the phases of the active season. An exogenous pulse of GnRH can apparently release behavior in male European ground squirrels, which is normally context dependent with the emergence of females.     

Serum prolactin levels and crop-sac development in ring doves during a breeding cycle

Using a turkey prolactin radioimmunoassay, the serum prolactin levels of male and female ring doves (Streptopelia risoria) during the breeding cycle were measured and their circulating prolactin levels were compared to crop-sac weight on a within-bird basis. During the early phase of the incubation period, crop weight showed a delayed response to prolactin stimulation and there was no correlation; during the midincubation period when prolactin and crop-sac weight were increasing, there was a strong positive correlation; around hatching, when prolactin was at its peak, there was no correlation. There were again strong positive correlations at later samples, when squabs were developing and both prolactin and crop-sac weight were declining. Thus, it appears that the correlation between the circulating prolactin level and crop-sac development depends on the stage of the breeding cycle. While males and females showed similar pattern of circulating prolactin during the period of incubation and parental care, only females consistently showed a postovulatory rise of prolactin. These results were discussed in the context of the role of prolactin in the breeding cycle.     

Proximity between the sexes in ring doves: Social bonds or surveillance?

Most bird species are monogamous, and in many instances the male provides a substantial contribution to the care of the young. Traditionally, the initial phase of the relationship between the sexes has been characterized in terms of pair formation and an increasing bond between the sexes. But recent formulations emphasize the role of guarding, surveillance, and other behavioural devices that protect the genetic relationship of the male to the offspring for which he cares. Ring doves were studied to determine whether the details of the relationship during the initial period are consistent with this latter view. Experiment 1 reveals that males remain in close proximity to their mates until the first egg is laid and the fertile period of the female ends; social contact then falls to a low level. Experiment 2 shows that when the female is removed during her fertile period, the male searches and calls more vigorously than does a female when her partner is absent. Finally, Experiment 3 demonstrates that it is visual contact rather than simply proximity that the male maintains during the fertile period of the female. These observations are consonant with the view that males maintain surveillance over their mates, but they do not exclude the possibility that the behaviour patterns might provide other benefits as well.     

Lesion-induced neurogenesis in the hypothalamus is involved in behavioral recovery in adult ring doves

Although neurogenesis in the brain of adult vertebrates is region dependent, lesion induces generation of new neurons in non-neurogenic brain regions. These findings raise the question of the role of new neurons in brain repair and functional recovery. We addressed this question by applying previous observations that electrolytic lesion induced neurogenesis in the ventromedial nucleus (VMN) of the hypothalamus in adult ring doves. Such lesions disrupted the male's courtship behavior, which could be reinstated after rehabilitation with a female. We investigated whether lesion-induced newborn neurons in the VMN facilitate the recovery of courtship behavior in the lesioned birds. We conducted systematic observations of cytological, morphological, and neuroanatomical changes in the lesioned VMN, and concurrently we monitored behavioral changes. Using a multitude of specific cell markers, we found a well-circumscribed cellular zone that proliferated actively. This highly proliferative zone initially appeared along the periphery of the lesion site, where cells had high levels of expression of neuronal, glial, and neurovascular markers. As newborn neurons matured at the lesion site, the necrosis gradually decreased, whereas a downsized proliferative zone relocated to a region ventral to the VMN. Some of the mature neurons were found to project to the midbrain vocal nuclei. Restoration of these projection neurons coincided with the recovery of courtship vocalization. Finally, we found that a social factor, that is, when the male doves were cohoused with a mate, facilitated neurogenesis and behavioral recovery. These results suggest that lesion-induced neurogenesis contributes to behavioral recovery in adult animals.     

Manipulation of male attractiveness induces rapid changes in avian maternal yolk androgen deposition

Avian eggs contain maternal androgens that may adjust offspringdevelopment to environmental conditions. We review evidenceand functional explanations for the relationship between androgenconcentrations in avian eggs and male attractiveness. Experimentalstudies in captive birds show generally positive relationships,but results from correlational and experimental field studiesare less consistent, perhaps because they lack a within-femaledesign to control for confounding between-female variation.We analyzed the effect of male attractiveness on yolk levelsof maternal androgens in a wild bird, using a correlationaland experimental approach with a within-female design. We manipulatedthe sexually selected UV coloration of the crown feathers ofmale blue tits (Cyanistes caeruleus) after their female hadlaid the second egg and measured the subsequent effect on androgenconcentrations (testosterone and androstenedione) in the fifth,seventh, and ninth eggs relative to that in the second egg.Levels of testosterone, but not androstenedione, in eggs 5 and7 were higher for control (attractive) than for UV-reduced (unattractive)males. This effect disappeared in the ninth egg, coincidingwith the recovery of UV coloration after manipulation. Thissuggests that females are capable of rapid adjustments of testosteronedeposition in response to changes in their mate's ornamentalplumage. However, androgen concentrations in the second eggand pretreatment male crown coloration were not correlated.Possibly, the combination of relatively small variation in UVcoloration before treatment and the influence of unknown confoundingvariables in the correlative approach resulted in insufficientstatistical power to detect such a correlation.     

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.     

Courtship raises male fertilization success through post-mating sexual selection in a spider

Courtship is well known for its positive effects on mating success. However, in polyandrous species, sexual selection continues to operate after copulation. Cryptic female choice is expected under unpredictable mating rates in combination with sequential mate encounters. However, there are very few accounts of the effects of courtship on cryptic female choice, and the available evidence is often correlative.Mature Argiope bruennichi females are always receptive and never attack or reject males before mating, although sexual cannibalism after mating occurs regularly. Still, males usually perform an energetic vibratory display prior to copulation. We tested the hypothesis that beneficial effects of courtship arise cryptically, during or after mating, resulting in increased paternity success under polyandry. Manipulating courtship duration experimentally, we found that males that mated without display had a reduced paternity share even though no differences in post-copulatory cannibalism or copulation duration were detected. This suggests that the paternity advantage associated with courtship arose through female-mediated processes after intromission, meeting the definition of cryptic female choice.     

Newborn GnRH neurons in the adult forebrain of the ring dove

The preoptic area of the hypothalamus is a key area that produces gonadotrophin-releasing hormone (GnRH). In birds, the chicken GnRH-I-form neurons are responsible for the hypothalamus-pituitary-gonadal system, which controls reproduction. In the ring dove, electrolytic lesion in the adult hypothalamus induces neurogenesis. In this study, we determined whether adult neurogenesis is involved in repairing GnRH neurons, specifically by generating newborn cells exhibiting GnRH-I immunoreactive properties. We selectively applied electrolytic lesions to three different regions of the diencephalon, including the preoptic area, which contains GnRH-I neurons, and identified new cells (BrdU-positive cells) that co-labeled with GnRH-I-immunoreactive cells. The BrdU⁺/GnRH⁺ double labeled cells were then confirmed with confocal laser analysis. In brains of both male and female ring doves we found new neurons at the lesion site of the preoptic region that were GnRH-I immunoreactive. However, the total number of GnRH neurons in the lesioned brains was less than that of sham-lesioned brains. When two other regions of the diencephalon that contain GnRH-I neurons were damaged, no recruitment of new GnRH-I neurons was detected. The rate of neurogenesis depends on the bird's reproductive phase when the lesion was applied. We found BrdU⁺/GnRH⁺ double-labeled cells almost exclusively during the pre-laying phase when birds are engaged in active courtship that leads to egg laying. Our observations suggest that recruitment of GnRH immunoreactive new neurons is restricted to the hypothalamic region and is sensitive to the reproductive stage of the birds.     

Lesion‐induced neurogenesis in the hypothalamus is involved in behavioral recovery in adult ring doves

Although neurogenesis in the brain of adult vertebrates is region dependent, lesion induces generation of new neurons in non‐neurogenic brain regions. These findings raise the question of the role of new neurons in brain repair and functional recovery. We addressed this question by applying previous observations that electrolytic lesion induced neurogenesis in the ventromedial nucleus (VMN) of the hypothalamus in adult ring doves. Such lesions disrupted the male's courtship behavior, which could be reinstated after rehabilitation with a female. We investigated whether lesion‐induced newborn neurons in the VMN facilitate the recovery of courtship behavior in the lesioned birds. We conducted systematic observations of cytological, morphological, and neuroanatomical changes in the lesioned VMN, and concurrently we monitored behavioral changes. Using a multitude of specific cell markers, we found a well‐circumscribed cellular zone that proliferated actively. This highly proliferative zone initially appeared along the periphery of the lesion site, where cells had high levels of expression of neuronal, glial, and neurovascular markers. As newborn neurons matured at the lesion site, the necrosis gradually decreased, whereas a downsized proliferative zone relocated to a region ventral to the VMN. Some of the mature neurons were found to project to the midbrain vocal nuclei. Restoration of these projection neurons coincided with the recovery of courtship vocalization. Finally, we found that a social factor, that is, when the male doves were cohoused with a mate, facilitated neurogenesis and behavioral recovery. These results suggest that lesion‐induced neurogenesis contributes to behavioral recovery in adult animals. © 2006 Wiley Periodicals, Inc. J Neurobiol, 2006     

Courtship in Drosophila quadrilineata with a unique male behavioral element, abdomen bending

This paper describes courtship of Drosophila quadrilineata of the immigrans group and reports a species-specific element, abdomen bending, that has not been described in Drosophila. The male bends his abdomen right and left as a display to the female. During courtship the male circles around the female, bends his abdomen, and taps her. The male licks the female ovipositor repeatedly, often after the females extrusion. Although the females ovipositor extrusion is thought to indicate rejection of a courting male in many species, the male of D. quadrilineata continues courtship. It is possible that the extrusion is a females mating signal to the courting male. The male of D. quadrilineata mounts the female in a rearward position and the genitalia of both sexes couple after mounting. This mounting position is shared by the species belonging to the immigrans group. We never observed behavior in which it appeared as if the male displayed the black stripes to the female. Males court one another and a courtship chain is frequently formed even in the absence of females. The males abdomen bending and the females extrusion followed by the males licking are discussed.