Male Sex Change Research Articles

Induction of sex change within two full moons during breeding season and spawning in grouper

Artificial sex change was induced within two full moons by an aromatase inhibitor (AI; 1 mg/fish) during the breeding season to establish the quickest method of sex change and natural spawning of the honeycomb grouper ( Epinephelus merra). The sex change from female (ovary) to male (testis) occurred during the time between the two full moons (3 weeks) following AI implantation, and the efferent ducts of sex-changed males were filled with sperm. To examine sperm fertility, sex-changed males were mated with natural, normal females and produced fertilized eggs. Most of the hatched larvae grew normally without any morphological deformities. Therefore, the use of this method which is the quickest known sex change method using AI, may contribute to quality sperm production for grouper aquaculture.

Induction of Female-to–male Sex Change in the Honeycomb Grouper (Epinephelus merra) by 11-ketotestosterone Treatments

The honeycomb grouper, Epinephelus merra, is a protogynous hermaphrodite fish. Sex steroid hormones play key roles in sex change of this species. A significant drop in endogenous estradiol-17beta (E2) levels alone triggers female-to-male sex change, and the subsequent elevation of 11-ketotestosterone (11KT) levels correlates with the progression of spermatogenesis. To elucidate the role of an androgen in sex change, we attempted to induce female-to-male sex change by exogenous 11KT treatments. The 75-day 11KT treatment caused 100% masculinization of pre-spawning females. Ovaries of the control (vehicle-treated) fish had oocytes at various stages of oogenesis, while the gonads of the 11KT-treated fish had transformed into testes; these contained spermatogenic germ cells at various stages, including an accumulation of spermatozoa in the sperm duct. In the sex-changed fish, plasma levels of E2 were significantly low, while both testosterone (T) and 11KT were significantly increased. Our results suggest that 11KT plays an important role in sex change in the honeycomb grouper. Whether the mechanism of 11KT-induced female-to-male sex change acts through direct stimulation of spermatogenesis in the ovary or via the inhibition of estrogen synthesis remains to be clarified.

Preventing behavioural interactions with a male facilitates sex change in female bluebanded gobies, Lythrypnus dalli

Sex change in marine teleost fishes is commonly regulated by social factors. In species that exhibit protogynous sex change, such as the bluebanded goby Lythrypnus dalli, the most dominant female typically initiates sex change when a male is removed from the social group. Females can use visual, chemical or tactile cues to assess the presence or absence of a male. The primary goal of our study was to determine whether the olfactory and visual presence of a male versus its behavioural interactions with females were important for mediating sex change. We exposed females to three different treatments: absence of a male, presence of a male that could physically interact with her and presence of a male behind a barrier that allowed visual and olfactory interactions but prohibited physical interactions. Sex change occurred in the absence of a male but not in the presence of a male that could physically interact with the female. The presence of a male behind the barrier did not prevent sex change but affected the timing of sex change. Season appeared to affect the latency to initiate male typical courtship, with a delay at the end of the reproductive season only when the male was present behind the barrier. We discuss the seasonal results in terms of L. dalli life history and the potential benefits and costs of changing sex late in the season in the presence or absence of aggressive reinforcement by the male. Our results identify direct behavioural interactions as an important proximate mechanism in the social regulation of sex change in L. dalli.

Changes in androgen‐producing cell size and circulating 11‐ketotestosterone level during female–male sex change of honeycomb grouper Epinephelus merra

11-ketotestosterone (11-KT), a potent male-specific androgen in fish, has important roles on spermatogenesis, male behavior, and nuptial coloration. The site of 11-KT synthesis and its role on male germ cell development during protogynous sex change is not clearly understood. We examined the dynamics of steroidogenic enzymes immunolocalization, viz cholesterol side-chain cleavage (P450scc), biomarker of steroids and cytochrome P45011beta-hydroxylase (P45011beta), downstream to 11-KT production, throughout the process of sex change in honeycomb grouper (Epinephelus merra). In female, P450scc immunoreactivity (-ir) was observed in the theca layer and tunica near blood vessels (BV). During the onset of sex change, P450scc reactive cells were observed in the remaining follicle layer of degenerated oocyte of the ovo-testis in early transitional (ET) and late transitional (LT). In male, P450scc-ir was localized in the interstitial Legdig cells of testis. P45011beta reactive cells were observed in the tunica near BV in female but not in theca layer. In ET and LT phases gonads, P45011beta localized in remaining follicle layer of degenerated oocyte and tunica near BV. On the other hand, in male, both interstices and tunica near BV showed strong signals against P45011beta. Moreover, in vivo and in vitro levels of 11-KT related with the changes in the nuclei diameter of P45011beta-positive cells in both tunica near BV and remaining follicle layer of degenerated oocyte to interstices during the progress of sex change. The present results suggest that 11-KT produced in the tunica near BV may provide the stimulus for female to degenerate oocytes and initiate sex change. However, 11-KT produced both in tunica near BV and remaining follicle layer of degenerated oocyte possibly plays critical role during testicular differentiation as well as gonadal restructuring at mid to late phases (ET to LT) of sex change in honeycomb grouper.

Changes in gonadal development, androgenic gland cell structure, and hemolymph vitellogenin levels during male phase and sex change in laboratory-maintained protandric shrimp, Pandalus hypsinotus (Crustacea: Caridea: Pandalidae)

Most pandalid shrimps show protandric hermaphroditism, and male sexual differentiation is considered to be controlled by the androgenic gland. In the present study, we examined the histology of gonadal development during the male phase and sex change and the involvement of the androgenic gland in regulating male reproduction in laboratory-maintained Pandalus hypsinotus. Juvenile shrimps developed testicular tissues in the peripheral part of gonads during the age of 16–31 months and produced spermatozoa between 34 and 36 months. After reaching sexual maturity, male shrimps exhibited seasonal testicular development: active production of spermatozoa (February–May), disappearance of spermatozoa (spent, April–June), increase of spermatocytes (May–November), appearance of spermatids and spermatozoa in the gonads (November–February). The androgenic gland cells became larger and the rough endoplasmic reticulum in the cytoplasm developed at male sexual maturity. The cell structure shows that the androgenic gland hormone is a peptide. Furthermore, bilateral eyestalk ablation on immature male shrimps induced hypertrophy of the androgenic gland and acceleration of male sexual maturation. These results indicate the involvement of androgenic gland hormone and some eyestalk factor in regulating male sexual maturation. Over a 1-year laboratory-rearing period, some male shrimps (16.7%) changed sex. In transitional shrimps, testicular tissues in the gonads and androgenic glands degenerated; on the other hand, oocytes started yolk protein accumulation and hemolymph vitellogenin levels became high. These results suggest that androgenic gland degeneration is a trigger for sex change and that the vitellogenin level is a useful marker for sex change.

Evidence that estrogen regulates the sex change of honeycomb grouper (Epinephelus merra), a protogynous hermaphrodite fish

Circulating estradiol-17beta (E2) levels decrease precipitously during female to male (protogynous) sex change in fish. Whether this drop in E2 levels is a cause or consequence of sex change is still largely unknown. The present study treated adult female honeycomb groupers (Epinephelus merra) with aromatase inhibitor (AI, Fadrozole), either alone or in combination with E2, to investigate the role of estrogen in protogynous sex change. Control fish had ovaries undergoing active vitellogenesis; the gonads of AI-treated fish had already developed into testes, which produced sperm capable of fertilization. In contrast, co-treatment of fish with E2 completely blocked AI-induced sex reversal. AI treatment significantly reduced circulating levels of E2, whereas the addition of E2 to AI prevented the loss. The plasma androgen (testosterone and 11-ketotestosterone) levels were increased in the AI-treated fish, while the levels in the E2-supplemented fish were low compared to controls. Present results show that E2 plays an important role in maintaining female sex of hermaphrodite fishes, and that the inhibition of E2 synthesis causes oocyte degeneration leading to testicular differentiation in the ovary.

Testing a new version of the size-advantage hypothesis for sex change: sperm competition and size-skew effects in the bucktooth parrotfish, Sparisoma radians

A variety of field studies suggest that sex change in animals may be more complicated than originally depicted by the sizeadvantage hypothesis. A modification of the size-advantage hypothesis, the expected reproductive success threshold model, proposes that sperm competition and size-fecundity skew can strongly affect reproductive pay-offs. Size-fecundity skew occurs if a large female’s fecundity is markedly higher than the aggregate of the other members of her social group and, together with paternity dilution from sperm competition, can produce situations in which large females benefit by deferring sex change to smaller females. Deferral by large females can create sex-size distributions characterized by the presence of large females and small sex-changed males, and it is precisely these distributions that the traditional size-advantage model cannot explain. We tested the predictions of the new model with the bucktooth parrotfish, Sparisoma radians, on coral reefs in St. Croix, U.S. Virgin Islands. Collections and spawning observations determined that the local environmental regime of S. radians is characterized by pervasive sperm competition (accompanying 30% of spawns) and factors that can produce substantial size-fecundity skew in social groups. Dominant male removal experiments demonstrate that the largest females in social groups often do not change sex when provided an opportunity. Instead, smaller, lower-ranking females change sex when a harem vacancy arises. This pattern of sex change is in contrast to virtually all previous studies of social control of sex change in fishes, but provides strong support for the general predictions of the expected reproductive success threshold model. Key words: coral reefs, Labridae, protogyny, Scaridae, seagrass beds, size-fecundity skew, social control of sex change, sperm competition. [Behav Ecol 15:129–136 (2004)]

Androgens stimulate sex change in protogynous grouper, Epinephelus coioides: spawning performance in sex-changed males

We examined the efficacy of androgens (1.0 mg/kg body mass), testosterone (T), 11-ketotestosterone (11-KT), 17α-methyltestosterone (MT), testosterone propionate (TP) or androgen mixture (T, MT and TP in an equal ratio), for induction of sex change in protogynous orange-spotted grouper, Epinephelus coioides. The spawning performance in sex-changed males was also investigated. MT and androgen mixture at a dose of 1.0 mg/kg BW induced a sex transition and completion of spermatogenesis up to the functional male phase. The androgen mixture was most effective. Significantly, higher plasma T levels were found in MT and androgen mixture groups compared to control and other androgen implantation (T, TP or 11-KT) groups. We found that plasma levels of estradiol-17β (E2) or 11-KT were not different among treated groups. Sex-changed males could successfully fertilize mature eggs. Fertilization and hatching rates were of 23.5–70.4% and 8.4–44.6%, respectively. The data demonstrated that induction of sex change by exogenous androgens in groups could apply to the aquaculture field for seed production.

Induced sex change, spawning and larviculture of potato grouper, Epinephelus tukula

The potato grouper ( Epinephelus tukula) is a new aquaculture species with high economic potential. This is the first report of induced sex change, reproduction and larval rearing in this species. The smallest body size at which mature females were observed was 90 cm in total length and 16 kg in body weight. The diameter of tertiary yolk globule stage of oocytes was 505±10 μm for 16–24 kg individuals. Larger oocytes (552±13 μm) were obtained from individuals at 24 kg body weight. Implantation with an androgen mixture (1000 μg/kg body weight) containing testosterone, 17α-methyltestosterone, and testosterone propionate in equal ratios, was applied to female groupers for the induction of sex change in March. Sex change had occurred by 30 days after androgen implantation, and the male ratio reached 66.7% by day 70. Natural mating and spawning activity were found in females and sex-changed males in April. The fertilized eggs were hatched and reared successfully with a series of live foods. Metamorphosis occurred and larvae became juveniles with spots and skin pigmentation by 29.3 mm body length, with complete reduction in the pelvic and dorsal fins by 33 days after hatch.

Male-male competition selects for delayed sex change in the protandrous shrimp Pandalus latirostris

In most protandrous species, male size advantage is generally regarded as unimportant in determining the timing of protandrous sex change. In pandalid shrimp, the size/age of male sex change often fluctuates among years and populations, but the adaptive significance of late reversing males (LRMs) is not well understood. This study experimentally examined the adaptive significance of LRMs in the protandrous pandalid shrimp Pandalus latirostris Rathbun. Field and laboratory studies were carried out in August–September of 1998–2002 on P. latirostris in Notoro Lagoon, Japan (44°03′N; 144°10′E). Mature females that had molted (i.e. mate receptive) were tethered in the field and their mating behavior with wild males was observed. Copulations occurred with a single male at a time, although other males could sequentially mate with a tethered female. Because tethered females rejected male approaches, males had difficulty transferring their spermatophores. In the laboratory, males copulated with non-tethered, recently molted females for only 15 min after molting. Recently molted females are wary of potential predators, since their soft exoskeleton makes them particularly vulnerable. Fast access by males enhanced fertilization success in this shrimp. The effect of male size on mating success in the laboratory was examined. Both small and large males successfully inseminated females in the absence of competitors. In experiments where large, medium, and small males competed for a female, however, larger males guarded females longer than smaller males, until the female molted and became receptive. Moreover, large males were more successful at copulating once molting occurred. These results imply that male-male competition drives delayed sex change in some situations.

The Humphead Wrasse, Cheilinus Undulatus: Synopsis of a Threatened and Poorly Known Giant Coral Reef Fish

The humphead wrasse, Cheilinus undulatus, is the largest living member of the family Labridae, with a maximum size exceeding 2 m and 190 kg. Its geographic range covers much of the Indo-Pacific. The species is not common, recorded maximum adult densities rarely exceeding 20 fish/10,000 m2. Small individuals are typically associated with high coral cover; larger fish are found mainly on outer or deep reefs, steep slopes and passes, singly or in small groups and seagrasses. However, for reproduction the species forms small spawning aggregations of tens to more than one hundred fish. The diet of the humphead wrasse includes large invertebrates and small fishes. The species attains at least 30 years and reaches sexual maturation at about 35–50 cm total length and <5 years of age. Most small adults are female while mainly males exceed 1 m and there is evidence of female to male sex change. The humphead wrasse is of considerable cultural value in some Pacific countries and is among the most prized in the live reef food fish export trade, for which it is often taken in its juvenile size range, either directly for sale or, increasingly, for grow-out to market size. It is also marketed chilled. The species is particularly sensitive to fishing pressure. In most fished areas, density and body size have dropped substantially. It appears to be particularly heavily targeted and depleted in SE Asia and in some places faces extirpation. The humphead wrasse is often taken by night spearfishing and by cyanide, with protection typically weak or non-existent, despite regulations and by a ‘vulnerable’ assessment on the 1996 IUCN Red List. The humphead wrasse has not been reared successfully in hatcheries. Other giant reef fish share many similar problems and detailed study of the humphead wrasse contributes to a better understanding and conservation of all such species. This review examines and evaluates published and gray literature, original unpublished research and correspondence with almost 50 knowledgeable workers. It examines the value of such sources for quickly, but adequately, assessing the conservation and management status and key data gaps in species that are little known, vulnerable, difficult and expensive to study and may require urgent management or conservation action.

Modeling spawning strategy for sex change under social control in haremic angelfishes

In haremic angelfishes where protogynous (female to male) sex change is favored, females have been reported to adopt several tactics for earlier sex change on the basis of a trade-off between reproduction and growth, or survivorship. A recent field study on Centropyge ferrugatus revealed that females reduce spawning frequency in competition with similar-sized neighbors for social dominance. To evaluate the optimal spawning strategy taken by haremic fishes, we developed an evolutionarily stable strategy model that focuses on their life history and social structure based on field data of C. ferrugatus. The results of the analysis predict that the spawning frequency will be low when the mortality rate of females is high compared with males, the harem size is large, and there is a moderate degree of social control. Our model further predicts conditions under which females completely stop spawning, as if they have become bachelors. Thus, the regulated spawning frequency may be taken as a strategy to optimize the reproductive success of an individual in response to the available choices for sex change, social control, and environmental conditions. Social control would also play an important role in sex change in many other haremic species.

P450scc-like immunoreactivity throughout gonadal restructuring in the protogynous hermaphrodite Thalassoma duperrey.

The source of steroid hormones, which potentially regulate gonadal restructuring throughout protogynous sex change in teleosts, remains largely unknown. To address this issue, immunocytochemical methods were employed to detect gonadal sites of steroidogenesis in the protogynous hermaphrodite wrasse Thalassoma duperrey at different stages in the sex change process. Steroidogenic cells were classified based on the presence of P450 cholesterol-side-chain-cleavage-like immunoreactivity (P450scc-ir). P450scc-ir cells were predominantly in the thecal layer of normal females. As females underwent sex change, P450scc-ir localization shifted from the thecal layer to the interstitium. P450scc-ir cells appeared to increase in number midway through sex change. In sex-changed males, P450scc-ir cells were found in small clusters interspersed among spermatogenic lobules. These results demonstrate for the first time the ability of the gonad to produce potential steroidal mediators of gonadal restructuring throughout the sex change process.

Immunocytochemical and Ultrastructural Identification of Pituitary Cell Types in the Protogynous Thalassoma duperrey during Adult Sexual Ontogeny

Abstract Protogynous wrasses (Thalassoma duperrey): females (F), primary males (PM) along with a few terminal-phase males (TM) and sex-changed males (SM), were used to characterize the topographical organization of the pituitary. In general, immunocytochemical and ultrastructural features of the adenohypo-physeal cell types of the saddleback wrasse pituitary resemble those of other teleosts. In the rostral pars distalis (RPD), corticotropic cells were found bordering the neurohypophysis (NH) and surrounding the centroventrally located prolactin cells. Thyrotropic cells formed a small group in the anteriodorsal part of the rostral and proximal pars distalis (PPD). The somatotropic cells were distributed in large clusters, mostly organized in cell cords around the interdigitations of the NH of the dorsal PPD. Cells containing gonadotropin Ib subunit were localized in the dorsal parts of the PPD, in close association with somatotropic cells and gonadotropin IIb subunit containing cells were seen in the centr...

Both-ways sex change in monogamous coral gobies, Gobiodon spp.

We confirmed both-ways sex change in the coral-dwelling gobies Gobiodon micropus, G. oculolineatus, G. quinquestrigatus and G. rivulatus rivulatus by mate-removal experiment in the field and by the aquarium experiment of keeping two consexual fish in a coral. Eight species of Gobiodon were found in Acropora corals on the reef flat of Sesoko Island, Okinawa, southern Japan. The 4 species mentioned above bred in monogamous pairs composed of a male and a female matched by size, and the male took care of eggs deposited on the coral branch. In G. quinquestrigatus and G. rivulatus rivulatus males were larger than females in newly formed pairs, and females grew faster than their mates until breeding. The growth-rate advantage in females seems to be the major factor in the evolution of female to male sex change. The gobies strongly depended on host corals, but they moved between the corals after mate loss or coral death to form new pairs. This provides opportunities for the evolution of male to female sex change; the ability to change sex in both directions reduces the frequency of risky movement between host corals to form new pairs. These conditions are very similar to those reported in the both-ways sex change of another coral-dwelling goby Paragobiodon echinocephalus.

Protogynous Sex Change in the Intertidal Isopod Gnorimosphaeroma oregonense (Crustacea: Isopoda).

In Crustacea, the dominant pattern of sequential hermaphroditism is protandry (sex change from male to female). Here we provide the first evidence from external morphology and population structure that Gnorimosphaeroma oregonense, an abundant, sexually dimorphic intertidal isopod, undergoes protogynous (female to male) sex change. In the field, 31% of females had rudimentary penes, suggesting sex change and laboratory growth experiments confirmed that females produced one brood of juveniles, then passed through a variable number of molts as immature males before becoming sexually mature males. Contrary to reports for other protogynous Crustacea, this study suggests that in G. oregonense sex change is not socially mediated, although it may be facultative, because a large percentage of laboratory-reared juvenile isopods developed directly into males. Potential adaptive explanations for protogyny are discussed in relation to protandry--the more common strategy in Crustacea.

Histological and ultrastructural evidence for the role of gonadal steroid hormones in sex change in the protogynous wrasse Thalassoma duperrey

The process of sex change in the protogynous wrasse, Thalassoma duperrey, was investigated through histological and ultrastructural observations on the gonads of females changing sex to male. Changes in plasma steroid levels concomitant with structural changes were measured by radioimmunoassay. The process of sex change from ovary to testis was divided into six stages on the basis of changes in the structure of the germinal and somatic elements. Ovaries of females were filled with vitellogenic oocytes during the breeding season, but contained no spermatogenic tissue (Stage 1). At the commencement of sex change (Stage 2), vitellogenic oocytes began to degenerate, and were ingested by macrophagous cells. This stage was accompanied by a rapid drop in plasma levels of estradiol-17β. Thereafter, previtellogenic oocytes (Stage 3) also began to degenerate, and aggregations of stromal tissue, and loose connective tissue were observed in the central region of the lamellae. Steroid producing cells (Leydig cells), developed at the border of this loose connective tissue. Presumed spermatogonia proliferated on the periphery of the lamellae, and Leydig cells increased in size and number (Stage 4). Spermatogonia formed cysts, and underwent spermatogenesis (Stage 5). Finally, sex change to male was considered complete, with the beginning of active spermatogenesis and spermiation (Stage 6). Plasma levels of testosterone remained low throughout the sex change, but a second androgen, 11-ketotestosterone increased gradually in parallel to the increased numbers of Leydig cells and spermatogonia. Preliminary in vitro incubation of gonads with salmon gonadotropin, revealed that sex-changed males had higher levels of 11-ketotestosterone production than did females, while females had higher levels of estradiol-17β production than did males. Production of both these steroids increased in a dose-related fashion with increasing doses of gonadotropin.

A comparison of the MMPI results for psychiatric patients and male applicants for transsexual surgery.

Previous studies have been evenly divided in concluding that male sex change applicants do or do not evidence signs of psychopathology on Minnesota Multiphasic Personality Inventory testing. To clarify the discrepant findings, Minnesota Multiphasic Personality Inventory scale comparisons were made between sex change applicants who had been living continuously as women, applicants who had been living predominantly as men, and groups of psychiatric inpatients and outpatients. Findings revealed that members of both applicant groups were more likely than psychiatric patients to score high on a measure of femininity. However, applicants living as men were as disturbed as psychiatric patients on all other measures of psychopathology, whereas those living as women showed a notable absence of psychopathology. Like the psychiatric patients, applicants living as men produced significant elevations on the Depression, Psychopathic Deviate, Psychasthenia, and Schizophrenia Scales. A variety of group comparisons also revealed serious emotional disturbance in one sex change applicant group and none in the other. The results help to explain the previous discrepancies in the literature and underline the importance of the "living as female" variable for predicting disturbance in sex change applicants.