Androgen and behavior in the male three-spined stickleback,Gasterosteus aculeatus. II. Castration and 11-ketoandrostenedione effects on courtship and parental care during the nesting cycle

Courtship declines and ceases while parental care increases in the presence of developing eggs during the nesting cycle of the male three-spined stickleback, Gasterosteus aculeatus. Furthermore, circulating 11-ketotestosterone (11KT) levels are higher during the initial "courtship phase" than during the later "parental phase," similar to that found in other paternal fishes. This study aimed to investigate a possible functional relationship between changes in 11KT levels and changes in reproductive behavior during the nesting cycle. To this end, groups of nonspawned and spawned male sticklebacks were sham-operated, castrated, or castrated and treated with 11-ketoandrostenedione (11KA), and the effects of the treatments on courtship and parental care were studied. Castration removed circulating 11KT, while 11KA replacement prevented the natural decline in 11KT during the parental phase (11KA converts to 11KT extratesticularly), as assessed by radioimmunoassay. Regardless of treatment, parental care remained low and courtship was present in all nonspawned males, even at the end of the experiment. However, courtship did eventually decline in castrated nonspawned males compared to the other two nonspawned groups. In all treatments of spawned males there was a drastic decline in courtship and an increase in parental care. In castrated spawned males, however, the decline in courtship came earlier than in the other two spawned groups. 11KA treatment did not prevent the natural decline in courtship/increase in parental care in spawned males, indicating that the natural decline in 11KT is not responsible for the main portion of the rapid changes in these behaviors over the stickleback's nesting cycle. The limited effects of castration also exclude other gonadal hormones from being responsible for most of these changes.     

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.     

Alternative reproductive tactics and courtship in the common goby

In the marine common goby Pomatoschistus microps , males either build nests and attract females to spawn or act as sneakers, forcing themselves into other males' nests during mating, to fertilize some of the eggs in the clutch. The reproductive tactics of small males were studied in the presence of surplus nest material and an excess of gravid females, while a larger, nestholding male spawned with a female. Of 24 small males, 13 only attempted to sneak, seven only courted females and/or spawned in their own nest, and four performed both behaviours. The small males who attracted females successfully were significantly larger than the ones who failed to do so. Females had been courting the small male more often when he subsequently spawned in his own nest, than when he did not get a female. The small males did not court the females very frequently, and their courtship rate did not depend on whether or not they attracted females. Instead, female courtship behaviour may be a cue for a male either to spawn as a nest holder or become a sneaker. The study pointed out the importance of female behaviour for male reproductive tactics, which could be more important in fish than is known currently.     

Differential effects of 11-ketotestosterone on dimorphic traits in a teleost with alternative male reproductive morphs

Species with dimorphic males typically have one morph (type I) associated with territoriality and courtship, and another morph (type II) associated with cuckoldry. In teleost fishes, type I males generally have higher levels of the androgen 11-ketotestosterone (KT) than type II males. Is KT causal to phenotypic differences between morphs? We investigated this question in the midshipman fish (Porichthys notatus) in which type I males have detectable levels of KT during the breeding season, whereas levels in type II males are usually undetectable. Type I midshipman will either cuckold or hold territories and court females, whereas type II males are only known to cuckold. Acoustic courtship by type I's is supported by the sonic motor nucleus, which innervates a sound-producing sonic muscle. Type I males have larger sonic motor nuclei and larger sonic muscles than type II males, consistent with the more dynamic vocal repertoire of type I's. Here, we tested whether intraperitoneal KT implants in adult type II males would induce type I male-like traits in brain, sonic muscle, and behavior. Type II's treated with KT did not differ from blank-implanted type II's in sonic motor nucleus volume. Sonic muscle mass increased in KT-implanted type II's, but did not reach the relative mass naturally observed in type I's. While neither territoriality nor courtship were induced, cuckoldry behavior intensified in KT-implanted type II's. Thus, for some but not all characters, KT exaggerated the expression of already existing type II male traits rather than inducing the type I male-like traits of territoriality, courtship, and an expansive vocal motor system.     

Steroid hormones and paternal care in the plainfin midshipman fish (Porichthys notatus)

The present study investigated the relationship between plasma steroid hormone levels and the expression ofpaternal behavior in the plainfin midshipman fish (Porichthys notatus), where males may simultaneously care for multiple clutches in different stages of development. Blood samples were collected from free-living parental males during that part of the breeding season when males may be found in various stages of parental care. Plasma 11-ketotestosterone levels were significantly higher in males with empty nests and nests containing only eggs than in males with nests containing embryos. All males with nests containing embryos had undetectable testosterone levels, whereas testosterone levels were detectable in many males with empty nests or nests containing only eggs. Estradiol levels were detectable in only a few males from nests with no eggs or nests containing only eggs. Cortisol levels were not correlated with stage of paternal care or with handling time. These results follow the frequently reported vertebrate pattern of declining androgen levels over the course of the breeding season or during the period of parental care. However, many male midshipman guarding nests containing only eggs had androgen levels similar to those of males whose nests contained no offspring. Thus the pattern of androgen levels exhibited by reproductively active parental male midshipman may reflect a compromise between investment in paternal care versus courtship and/or territoriality.     

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.     

Time spent close to a sexual partner as a measure of female mate preference in a sex-role-reversed population of the blenny<Emphasis Type="Italic"> Salaria pavo</Emphasis> (Risso) (Pisces: Blenniidae)

In most species females do not exhibit conspicuous sexual behaviours and female mate preferences are often measured by means of the time spent close to males. In spite of its widespread use, in only a few studies has this measure been validated as a reliable indicator of female mate preference. Sex-role-reversed species offer a better opportunity to test female preferences, as females usually court males. We tested in a sex-role-reversed population of the blenny Salaria pavo (Risso) if the time spent by females close to males related to the number of courtship displays directed towards males, and whether these measures of female preference reliably predicted matings. Females were simultaneously presented with two males behind a glass partition and the time spent close to each male was measured. We then allowed females to have access to the males' compartments and measured the courtship behaviours performed by females towards each male and recorded with which of the males females spawned. Females spent more time close to the male that subsequently received more courtship displays, and the preferred male had a more developed head crest. However, both measures of female preference failed to predict matings. Females were often attacked by males and probably had, in some occasions, to spawn with the less preferred male. Females that spawned with the previously preferred male had more swollen bellies than females that spawned with the less preferred male, suggesting a male preference towards more ripe females. These results validate the use of "time spent close to a sexual partner" as a measure of female preference in S. pavo. We argue that matings may not always reliably indicate mate preferences, particularly in the less choosy sex.Communicated by R. Serrão Santos     

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.     

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.     

Male-male competition and female choice in brown trout

In some salmonid species, the females have been assumed to choose their mates on the size of the male's adipose fin. This hypothesis was tested in a stream water aquarium, in which 19 brown trout, Salmo trutta, females were allowed to choose between two males of the same body size but with different adipose fin sizes. The two males were separated from each other in cages. After the female had started to prepare her nest close to one of them, the males were released and allowed to fight each other for the opportunity to spawn. Out of 19 females, 14 prepared a nest closest to the male with the larger adipose fin. However, only six of the 14 females spawned with this male. Males that spawned were more dominant (i.e. were more likely to win fights). When the female spawned with the male she chose, he was less aggressive towards her than when she spawned with the other male. There were no significant differences in the plasma levels of testosterone (T) and 11-ketotestosterone (11-KT) between the chosen males and those not chosen. However, the dominant males had significantly higher plasma levels of T and 11-KT both before and after the experiment. The results support the view that female brown trout exhibit mate choice, but their choice is overruled by male-male competition. Copyright 1999 The Association for the Study of Animal Behaviour.     

Androgens and male mating behavior in rough-skinned newts, Taricha granulosa

Cyproterone acetate was administered either orally or intraperitoneally to intact, adult male newts, Taricha granulosa. The number of males that exhibited the courtship behavior of clasping when tested with nuptial females was not altered by the antiandrogen treatments. In males which were unresponsive to nuptial females, the occurrence of clasping was not evoked by injections for 4 days of testosterone, dihydrotestosterone, or 11-ketotestosterone. Further, the incidence of clasping was not significantly elevated by injections of prolactin and/or testosterone for 30 days. The effect of sexual activity on testosterone and dihydrotestosterone levels in male newts was determined by radioimmunoassay of plasma collected from males which were: (1) isolated from females; (2) allowed to clasp a female for 2 min; or (3) allowed to clasp a female for 1 hr. The testosterone and dihydrotestosterone levels were unchanged during this period of clasping. In February and again in June, plasma androgen concentrations were measured in males which differed in their propensity to initiate courtship when paired with females. Androgen levels were similar for males that clasped a female and males that never attempted to clasp a female. Plasma androgen levels in the male newt are apparently not correlated with sexual responsiveness.     

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.     

Effect of clutch size on male egg-fanning behavior and hatching success in the goby, Eviota prasina (Klunzinger)

In fish that exhibit paternal care, the females often choose their mates on the basis of male traits that are indicative of the parental ability of the males. In a marine goby, Eviota prasina, males tend their eggs within their nests until hatching, and females prefer males that have longer dorsal fins and exhibit courtship behavior with a higher frequency as their mates. In order to clarify the relationship between these sexually selected traits and the parental ability of males of E. prasina, the factors affecting the hatching success of eggs within male nests and the male parental care behavior were examined in an aquarium experiment. Females spawned their eggs in male nests and the clutch size of females showed a high individual variation (range = 88-833 eggs). The hatching success of eggs within male nests showed a positive correlation with the time spent by males in fanning eggs and the clutch size. In contrast to the prediction, however, the hatching success did not show a significant correlation with the sexually selected traits, i.e., the male dorsal fin length and the frequency of courtship displays. Moreover, multiple regression analysis indicated that the time spent by the males in fanning was the most important factor affecting the survival rate of the eggs. The time spent by males in fanning behavior was influenced by the clutch size within their nests; the fanning behavior of males occurred with a higher frequency when they tended larger clutches. Males are required to invest a greater effort in egg-tending behavior to achieve a higher hatching success when they receive larger clutches, probably due to the greater reward for their parental behavior. Based on their mate choice, females may obtain other benefits such as high quality offspring.     

Mate copying versus preference for actively courting males by female Japanese medaka (Oryzias latipes)

We determined whether female Japanese medaka (Oryzias laiipes)copied the choice of other females or independently preferredactively courting males. We allowed focal females to observeand subsequently choose between a male that courted and potentiallyspawned with a receptive female and a male that courted andwas rejected by an unreceptive female. When the receptive femaledid not spawn, the focal female preferred the male with thehigher courtship rate; whereas when the receptive female spawned,the focal female preferred the spawning male. Our results suggestthat female medaka prefer actively courting males, unless theyhave the opportunity to copy the mate choice of another female.     

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.     

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.     

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.     

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.     

Changes in male courtship intensity and androgen levels during brood cycling in the blenniid fish Rhabdoblennius nitidus

Androgens, for example testosterone, are major hormones that affect male courtship activity, territorial activity, sexual dimorphism, and reproductive tactics; they reach, and are maintained at, a sufficient level to express such sexual traits at the appropriate time for reproductive success. This study examined the effects of androgen levels (testosterone and 11-ketotestosterone) on male brood cycling with two distinct reproductive phases (i.e. courtship and parental phases) of a paternal brooding blenny Rhabdoblennius nitidus. Our study showed that time spent on courtship behaviour and androgen levels decreased with progress of brood cycling. In addition, time spent on courtship behaviour of males administered cyproterone acetate, an anti-androgen, was shorter than that of control males. These results indicate that the brood cycling is affected by the change in androgen levels. The study also showed that androgen levels decreased after acquisition of the eggs, irrespective of available spawning space in the nests. This result suggests that the presence of eggs themselves may be a trigger for the shift from the courtship phase to parental phase.