Androgen and behavior in the male three-spined stickleback,Gasterosteus aculeatus I.--changes in 11-ketotestosterone levels during the nesting cycle

Circulating 11-ketotestosterone (11KT) levels are higher during the courtship phase than during the later parental phase in a number of male teleosts. The present study describes the temporal changes in 11KT levels and their relationships to changes in courtship behavior, after different number of spawnings, over the nesting cycle of the male three-spined stickleback, Gasterosteus aculeatus, a small teleost showing pronounced paternal behavior. Plasma 11KT levels, measured by radioimmunoassay, were approximately 34 times higher during the initial courtship phase than at the end of the following parental phase in spawned males. In addition, males that spawned with three or more females on a single day showed an earlier decline in 11KT levels and in courtship behavior compared to males that were only allowed to spawn with a single female. Plasma 11KT levels remained high in males not allowed to spawn.     

Changes in reproductive physiology and behaviour over the nesting cycle in male three‐spined sticklebacks

Plasma 11‐Ketotestosterone (11 KT) and testosterone (T) levels and spiggin‐mRNA levels, as well as the kidney‐somatic index ( I _K) were measured in sexual males and in paternal males at the middle (5 days paternal) and at the end (8 days paternal with hatched eggs) of the nesting cycle in three‐spined sticklebacks Gasterosteus aculeatus from two populations. Glueing (using threads of 11 KT induced kidney‐protein spiggin) and fanning behaviour was measured daily. Fanning increased in paternal fish and remained low in sexual males. Plasma 11 KT and T levels, on the other hand, declined significantly in parental compared to sexual males as did spiggin expression, I _K and glueing behaviour. Thus, the drastic decrease in circulating 11 KT levels during the later parental phase may have resulted in an energy‐saving decrease in spiggin‐production and glueing, when this was no longer needed for nest maintenance. In addition, the mRNA levels of the β‐subunits of both gonadotropins, luteinising hormone (LH) and follicle stimulating hormone (FSH) were measured. The expression of both gonadotropins declined in the parental phase (not significant for β‐FSH in one of the populations) which was consistent with a decline in androgen levels possibly controlled via decreased gonadotropin secretion.     

Plasma levels of androgens and cortisol in relation to breeding behavior in parental male bluegill sunfish, Lepomis macrochirus

Like many teleosts, male bluegill sunfish (Lepomis macrochirus) provide sole parental care. To understand some of the proximate costs of parental care, we measured body condition and plasma levels of testosterone (T), 11-ketotestosterone (11KT) and cortisol in nesting bluegill males during pre-spawning, spawning and parental care stages. T and 11KT were at their highest mean levels during the pre-spawning period and decreased to lower levels early during the parental care period before rising again when the eggs hatched. Cortisol levels fluctuated across the breeding stages, but there was a noticeable increase from low levels on the day of spawning during the first 2 days of parental care when egg fanning is most intense. Levels of all hormones varied considerably among males, with androgen levels often correlating positively with a male's body condition. We also demonstrate, using a brood reduction experiment and repeated sampling of known individuals, that the presence of eggs affects hormone levels shortly after eggs hatch. Parental males in better body condition had higher levels of androgens during parental care. Males that were known to renest later in the season also had higher androgen levels and were in better body condition during the first nesting bout than males only known to have nested once. However, circulating levels of cortisol did not differ significantly between these groups. We discuss our findings in the context of proximate and ultimate costs of parental care and propose several reasons why elevated androgen levels may not be as incompatible with the expression of paternal care in male teleosts, as compared with avian and mammalian fathers.     

Egg fertility and prolactin as determinants of reproductive recycling in doves

When a pair of male and female ring doves in breeding condition is introduced in a breeding cage containing a nest bowl and a supply of nesting material, a regular and predictable sequence of behavioral changes can be observed. A complete breeding cycle consists of courtship and nesting (7–10 days), incubation (14–15 days), and parental care (21 days). A new cycle (recycling) starts following the departure of the fledglings. This study concerns itself with the occurrence of recycling before completion of a breeding cycle, with a special reference to the fertility of eggs. The fertility of eggs before and after oviposition regulates the timing of recycling. Laying infertile eggs as a result of having mated with impotent males (castrated + oil or castrated + testosterone proprionate) and then sitting on such eggs led to a 100% recycling with the shortest latency. In contrast, no doves recycled in the normal breeding pairs (females paired with intact males) until eggs were hatched and squabs fledged. For bird laying fertile eggs and sitting on infertile eggs or vice versa, half recycled, while the other half did not. The possibility that prolactin plays a role in mediating the onset of recycling was tested, and the significance of the results was discussed.     

Courtship behavior of male white perch, Morone americana: evidence for control by androgens.

Courtship behaviors are androgen-dependent in many vertebrates and castration often decreases courtship. We examined the effectiveness of castration in reducing courtship behaviors and 11-ketotestosterone (KT) and testosterone (T) in restoring them in male white perch. Castrates were given implants containing KT, T or no hormone. Sham-operated males received implants without hormone. Three weeks later, males were exposed to an ovulated female for 1 h and two courtship behaviors were quantified. Attending behavior involves close and continuous following of a female with occasional contact. Circling involves rapid transits around the female in a circular pattern or back and forth in front of her. In plasma samples taken immediately after observations, KT and T were below detectable levels in castrated males but at high physiological levels in males implanted with KT or T. Castrated males given KT attended females more than castrated males given T implants or implants containing no hormone, but not more than sham-operated males. Circling was eliminated by castration but restored by implantation with T or 11-KT to values exhibited by sham-operated males. This is one of the few demonstrations that KT can regulate courtship behavior in a non-territorial and economically important fish species.     

Courtship behavior of male white perch, Morone americana: evidence for control by androgens

Courtship behaviors are androgen-dependent in many vertebrates and castration often decreases courtship. We examined the effectiveness of castration in reducing courtship behaviors and 11-ketotestosterone (KT) and testosterone (T) in restoring them in male white perch. Castrates were given implants containing KT, T or no hormone. Sham-operated males received implants without hormone. Three weeks later, males were exposed to an ovulated female for 1 h and two courtship behaviors were quantified. Attending behavior involves close and continuous following of a female with occasional contact. Circling involves rapid transits around the female in a circular pattern or back and forth in front of her. In plasma samples taken immediately after observations, KT and T were below detectable levels in castrated males but at high physiological levels in males implanted with KT or T. Castrated males given KT attended females more than castrated males given T implants or implants containing no hormone, but not more than sham-operated males. Circling was eliminated by castration but restored by implantation with T or 11-KT to values exhibited by sham-operated males. This is one of the few demonstrations that KT can regulate courtship behavior in a non-territorial and economically important fish species.     

Androgen levels and social interactions in breeding males of the peacock blenny

Nest-holder male Salaria pavo have lower circulating concentrations of 11-ketotestosterone (KT) at the beginning of the breeding season than at its peak. At that peak density of nesting males was higher as were the number of visits of sneaker males to nests and of agonistic interactions between nest-holders and sneaker males. There was no difference between the two dates either in the frequency of male-male interactions or in the frequency of courtship episodes. Thus, higher plasma levels in nest-holders might be explained by a more intense sneaking pressure at the peak of the breeding season. At that peak, nest-holders had higher plasma levels of KT and a higher testosterone (T) to KT metabolization index in the gonads than did floater males. Both nest-holders and floaters had higher levels of KT and T in the testicular gland than in the testis. The levels of both androgens in the testicular gland, but not in the testis, were correlated with circulating concentrations of KT. These results suggest that the testicular gland is the major source of circulating KT in blenniids. Nest-holders had higher metabolization indexes than floaters both in the testis and in the testicular gland, which suggests that nest holding status promotes the conversion of T into KT.     

Unravelling the evolution of complex reproductive traits with phenotypic engineering

The social dynamics surrounding courtship, mating and parental care are complex enough when just a single male and female are involved, but for species that employ multiple strategies for achieving fertilization success, the network of interactions among rivals, allies and suitors can be utterly complicated. Such is the case in the ocellated wrasse, Symphodus ocellatus, in which males adopt one of three mating strategies. The large, colourful “nesting males” court females, defend territories and care for fertilized eggs until they hatch. The smaller “satellite males” help the nesting males court females and guard against the third morph, the “sneaker males”, which sneak in when a nesting male is spawning with a female and surreptitiously release sperm. Sneaker males perform no courtship displays nor defend territories, so their reproductive investment is devoted entirely to sperm production. And these alternative male strategies work: 100% of nests contain some eggs fertilized by sneaker and satellite males, despite the fact that parental care is solely the responsibility of nesting males In this issue of Molecular Ecology, work to untangle the proximate mechanisms regulating the reproductive physiology of nesting males and their behaviour towards other males, which impacts the entire social network. Moreover, they describe how variation in neuroendocrine regulation can give rise to variation in reproductive traits, upon which sexual selection can act.     

Elevated 11-ketotestosterone during paternal behavior in the Bluebanded goby (Lythrypnus dalli)

The relationship between androgens and paternal behavior is not straightforward, potentially because of the diversity of tasks a male must undertake to maximize reproductive success, notably alternating between courtship, aggression, and offspring care. In some species, these events are separated in time, but in others they are coincident. The endocrine profiles of species that simultaneously court, parent, and defend a nest, such as male bluebanded gobies (Lythrypnus dalli), are not well understood. We sampled a potent fish androgen, 11-ketotestosterone (KT), at different life history stages (experienced parenting males, experienced males not actively parenting, inexperienced males with their first clutch, and females), to examine this relationship. We found that experienced parenting L. dalli males have the highest KT levels of any group, while none of the other groups differed significantly. Males showed elevated KT levels when they had eggs compared to when they did not. Our data suggest that KT facilitates at least some aspects of parental care in L. dalli.     

Paternity, parental behavior and circulating steroid hormone concentrations in nest-tending male bluegill

Like many teleost fishes, bluegill (Lepomis macrochirus) are characterized by sole male parental care of offspring. In addition, bluegill parental males experience cuckoldry by specialized parasitic male morphs. This cuckoldry has previously been shown to influence the expression of parental care behavior. To better understand some of the proximate mechanisms mediating parental behavior, we examined the relationships between circulating steroid hormones, paternity, and parental behavior during the egg and fry stages of care in parentals that spawned during the first third of the breeding season. During the egg stage of care, we found that males with higher paternity had lower levels of testosterone, but there was no relationship between paternity and either 11-ketotestosterone or cortisol. There also was no relationship between the hormones and care behavior comprising fanning of the eggs, nest rim circles, chases of brood predators, or pecking at the eggs (indicative of egg cannibalism), except for a negative relationship between cortisol and pecking behavior. During the fry stage of care, we conversely found that males with higher paternity had higher levels of testosterone and 11-ketotestosterone. There also was a negative relationship between the concentrations of these two androgens and the defensive behavior of males when exposed to a potential brood predator (a pumpkinseed, Lepomis gibbosus). We discuss these results in relation to previous work in fishes and other vertebrate taxa. Overall, our data suggest a complex relationship between circulating steroid hormone levels, paternity and parental behavior.     

Endocrine control of sexual behavior in sneaker males of the peacock blenny Salaria pavo: effects of castration, aromatase inhibition, testosterone and estradiol

The effects of castration and sex steroid manipulations on the expression of sexual behavior were investigated in a small fish, the peacock blenny, Salaria pavo. In this species, large males defend nests and attract females while small "sneaker" males reproduce by imitating the female morphology and courtship behavior in order to approach nests during spawning events and parasitically fertilize eggs. Sneakers switch into nest holders in their second breeding season, thus displaying both male and female-like sexual behavior during their lifetime. We tested the effects of castration and of an aromatase inhibitor (Fadrozole, F), testosterone (T) or 17beta-estradiol (E(2)) implants on the expression of male and female-like behavior in sneakers. Sneakers were either sham-operated, castrated or castrated and implanted with vehicle, F, T+F or E(2)+F. Seven days after the treatment, sneakers were placed in a tank with a nesting male, two ripe females and an available nest. Castrated fish had lower levels of circulating T and increased the time spent displaying female typical nuptial coloration. T implants had the opposite effect, inhibiting the expression of female-like behavior and coloration. E(2) implants had no significant effect on the display of sexual behavior but the frequency of aggressive displays decreased. The results agree with previous findings in sneakers of S. pavo that demonstrated an inhibition of female-like behavior by 11-ketotestosterone (11-KT). The reported increase in T and 11-KT production when sneakers change into nest holders may thus contribute to behaviorally defeminize sneakers. Contrarily, both T and E(2) failed to promote male-like behavior, suggesting that behavioral masculization during tactic switching depends on other neuroendocrine mechanisms or that the time length of the experiment was insufficient to induce male-like behavioral changes in sneakers.     

Territorial behaviour of the male johnny darter,Etheostoma nigrum

Synopsis A field study was undertaken to describe the territorial behaviour of male johnny darters (Etheostoma nigrum) over the reproductive cycle. While defending eggs, males performed fewer Lateral Displays and lost fewer encounters with conspecific males than in the pre-egg phase. As the reproductive cycle progressed males were less responsive to females and performed relatively fewer courtship and more aggressive displays. Nesting males apparently shifted from a sexual to a parental phase shortly after acquiring eggs.Frequency of response, responsiveness (number of responses/number of intrusions x 100) and maximum distance of response (MDR) of male johnny darters were highest early in the reproductive cycle and decreased over the egg-guarding phase. The patterns of territorial defence exhibited by male johnny darters over the reproductive cycle were similar to those of other species of fish with paternal care.     

The Influence of Parental Status on Courtship Effort in a Paternal Caring Fish

It is widely assumed that caring for young limits the motivation of parents to seek additional mating opportunities. However, in situations where parental care does not involve direct provisioning of the offspring, but rather activities directed at the brood as a whole (e.g. guarding), it may be more efficient for parents to care for large numbers of young at once. This may be especially true for species with exclusive paternal care, with fathers that have recently acquired a brood of young potentially benefitting from vigorously courting prospective mates, so as to maximise their chances of attaining a large number of young to rear together. We experimentally tested this hypothesis in the three‐spined stickleback (Gasterosteus aculeatus), a fish with male only care. Contrary to our predictions, we found no evidence of any differences in courtship between recently spawned egg‐tending fathers and males that had not spawned. However, males that were permitted to spawn, but then had their eggs taken from them, courted less vigorously. Together, the results of our study suggest that the potential benefits of vigorous courtship in terms of acquiring additional young may be offset by additional costs faced by parental males.     

Multiple mechanisms of phenotype development in the bluehead wrasse

Despite having detailed information on mechanisms mediating sex-typical behavior in many species, we have little understanding of whether the same mechanisms regulate these behaviors when they are performed in the same species under different social contexts. In the five field experiments of this study of bluehead wrasses (Thalassoma bifasciatum), a sex-changing fish, we examined the roles of arginine vasotocin (AVT) and the potent teleost androgen 11-ketotestosterone (11KT) in mediating sexual and aggressive behaviors typical of dominant males. We demonstrated that AVT appears necessary for the assumption of dominant territorial status in males and females, but is sufficient only in the socially dominant terminal phase (TP) male phenotype. Specifically, an AVP V(1) receptor antagonist prevented both TP males and females from gaining dominance over recently vacated territories. However, unlike TP males in a previous study, neither females nor initial phase males responded to AVT treatment with increases in display of TP male typical behaviors when under social conditions that inhibit sex change. Treating females with 11KT did not alter responsiveness to AVT, but did induce male coloration and courtship behavior that was not observed in oil-treated females. Combined with the results of a previous study, these results indicate that the ability of AVT to induce male-typical behavior differs among sexual phenotypes and that this differential responsiveness appears to be dependent on social context and not directly on exposure to 11KT. Furthermore, since 11KT can induce courtship behavior in females that is not affected by AVT, there may be different hormonal mechanisms mediating courtship behavior under different social contexts.     

Psychobiology of sexual behavior in a whiptail lizard, Cnemidophorus inornatus

This study investigated the effects of social environment on gonadal recrudescence and sexual behavior in male and female Little Striped Whiptail lizards (Cnemidophorus inornatus). The presence of sexually active males facilitates ovarian recrudescence in conspecific females. Similarly, the presence of reproductively active females facilitates testicular recrudescence in conspecific males. Males housed with females, however, had lower average circulating concentrations of testosterone and dihydrotestosterone, and higher average concentrations of corticosterone compared to intact males housed in isolation. In other studies, the presence of reproductively active females partially restored courtship behavior in castrated males compared to castrated males housed in isolation. Despite the stimulatory effects of females on castrates, exogenous androgens are required for complete restoration of all components of sexual behavior in male C. inornatus. Females are receptive to male courtship and copulatory behavior only during the vitellogenic stages; females in previtellogenic or postovulatory ovarian stages aggressively reject male courtship advances. These findings demonstrate reciprocal effects of sexual behaviors of males and females upon each other's reproductive behavior and physiology.     

Size-Based Variation in Somatic Energy Reserves and Parental Expenditure by Male Smallmouth Bass, Micropterus dolomieu

Male smallmouth bass show size-based variation in both probability and timing of reproduction. The objective of this research was to determine seasonal and size-based patterns of depletion of energy reserves and determine if parental defense is related to males' energy reserves. We sampled male smallmouth bass in the spring, during the parental care period and in the fall to measure energy reserves (lipid stores in muscle and viscera tissue) over a two year period. Energy stores, which were not built up before nesting, declined to a minimum level by the end of the parental care period. Small males had consistently lower energy reserves than larger males and did not utilize these reserves at the same rate during the parental care period. All parental males complimented endogenous energy reserves by feeding during parental care, however, small males appear to rely proportionately more on exogenous energy intake than do larger males. Parental defense by all sizes of males declined over the parental care period, the decline being the most obvious by small males. Small males' lower energy budget may make them less effective parents and decrease their probability of survival over the following winter relative to larger males.     

Sex-role reversal in the tidewater goby, Eucyclogobius newberryi

Species in which females compete more intensely than males for access to mates are uncommon. Sex-role reversal in fishes has been documented only in species in which males bear eggs, such as pipefish and a mouth brooding cardinalfish. I investigated the reproductive behavior of the tidewater goby, Eucyclogobius newberryi (Gobiidae), to determine whether and to what degree this species is sex-role reversed. Males constructed and defended burrows for spawning in sand. Both sexes initiated courtship, but the female's breeding coloration was more striking. The intensity of sexual aggression was greater among females than among males. The female laid her entire clutch with a single male, and the male accepted only one clutch per brooding cycle. Both sexes spawned repeatedly (up to 12 times in aquaria), but fish did not form permanent pairs. Males cared for eggs in the burrow 9–11 days until hatching, and rarely if ever emerged to feed. Many aspects of male behavior (nest construction and defense, courtship, and parental care) were typical of most gobiids. On the other hand, female behavior (black nuptial coloration and intense female-female competition) was unusual, not only for gobiids but for animals in general. I therefore concluded that the tidewater goby is moderately sex-role reversed. Its sexual behavior is apparently unique among fishes because it is the only reported case of sex-role reversal in teleost males that do not bear eggs or developing young. This revised version was published online in July 2006 with corrections to the Cover Date.     

Temporal patterns of circulating LH and steroid hormones in a brood parasite, the Brown-headed cowbird, Molothrus ater.

The reproductive cycle of monogamous passerines is characterized by a sequence of events leading from courtship and pair formation through nest building, egg laying and incubation, and ending with the care of young. Each of these stages is accompanied by equally predictable changes in the pattern of hormone secretion. This report describes endocrine changes in the plasma of free-living Brown-headed cowbirds, Molothrus uter , during a breeding season. Changes in some morphological parameters are also presented.
Cowbirds are brood parasites that show no parental behaviour and lay eggs almost daily for approximately eight weeks. Males are not territorial, but they mate guard extensively. Hcncc, their breeding biology is quite different from that of the nesting species described above. They are hormonally distinct, as well. Luteinizing hormone rises in the spring and remains elevated throughout May and most of June. By July, when breeding has ceased, plasma luteinizing hormone levels are at pre-breeding values. Plasma androgens are also maintained at peak levels for an extended period of time, although they decline briefly between the establishment of dominance hierarchies and the onset of breeding. Circulating levels of corticosterone are elevated until the middle of the breeding season, when they begin to fall, although here, too, there is a transient depression just prior to breeding. In the light of the studies on nesting species, these data suggest that hormone profiles are closely correlated with the breeding and social activities that characterize the species under investigation.     

Reproductive history and nest environment are correlated with circulating androgen and glucocorticoid concentrations in a parental care-providing teleost fish

Using a long-term study population of wild smallmouth bass Micropterus dolomieu in a connected river-lake system, we investigated whether circulating glucocorticoid (cortisol) and androgen (testosterone) concentrations are influenced by reproductive investment and nesting environment in fish providing nest-guarding paternal care. For all individuals, we collected measures of reproductive history and the value of current parental care. We assessed nest environment and monitored individuals to quantify seasonal reproductive success. Finally, we measured circulating cortisol concentrations following a standardized stressor and circulating testosterone concentrations. Using general linear models, we found that poststress circulating cortisol concentrations were positively related to water temperature and were higher in fish nesting in the river than in the lake. Circulating testosterone concentrations were negatively related to water temperature and were higher in reproductively experienced fish. When considering the factors that influence reproductive success, we found that only parental size was positively related to current nest success. In summary, the results demonstrate that nesting environment is correlated with parental stress responses during parental care, while reproductive history and nesting environment are correlated with circulating androgen concentrations. Collectively, these results offer insight into the roles of both glucocorticoid and androgen steroid hormones during parental care in teleost fish.