Human chorionic gonadotropin (hCG) induces gonad reversal in a protogynous fish, the bluehead wrasse, Thalassoma bifasciatum (Teleostei, Labridae)

Female to male successive hermaphroditism (protogyny) is common in several groups of marine fish. Thalassoma bifasciatum, the bluehead wrasse (Labridae), found in the reefs of the Caribbean normally undergoes sex reversal after receiving behavioral cues. This report deals with the successful use of human chorionic gonadotropin (hCG) in inducing gonad reversal in this species. Eighty percent (n = 40) of the treated fish showed signs of reversal in 1-6 weeks; only 11% (n = 54) of the control (nontreated) group showed signs of reversal during the same period. The number of fish undergoing reversal increased with the length of the treatment period, 55% after 1 week, 100% after 6 weeks. A bluehead color pattern, typical of the terminal male phase, also appeared more frequently in the longer treated groups. To determine the efficacy of hCG in inducing gonad reversal, morphological criteria for reversal had to be established. For the majority of treated fish, the presence of both spermatogenic cysts and degenerating oocytes was sufficient to label these gonads unambiguously as undergoing reversal. However, at the extremes of this process, i.e., onset (early) and endpoint (late) stages, ambiguity could arise in identifying a gonad as undergoing reversal, and, therefore, key criteria were established for these stages. In our hands, the most consistent and reliable criterion for the early stage reversal was appearance of male germ line cells, clusters of "spermatogonial-like" cells. For the late stage, recognizable remnants of late-state oocytes had to be present. Some details of the histological changes that occur during early, middle, and late stages of reversal are also described. It is suggested that these results with hCG shed new light on the endocrine control of gonad reversal in T. bifasciatum.     

The feeding strategies of a facultative cleanerfish, Thalassoma bifasciatum (Pisces: Labridae)

The adult Bluehead wrasse (Thalassoma bifasciatum) is a facultative cleanerfish that can change its colouration into any one of three: patterns. These patterns correspond to several modes of feeding behaviour which, in turn, can be correlated with the movements of host fishes. The barred pattern occurred on individuals that are far ranging and also on those that formed unstable cleaning groups. These groups tended to form at locations having sharp drops in depth of several metres. Here these groups serviced large groups of host fishes. The striped pattern occurred on solitary individuals with a limited home range. They occurred in very shallow areas and serviced small groups of host fishes. The bright yellow pattern developed when a large food source was discovered and may be related to the attraction of conspecifics.     

The effects of testosterone implants on ovarian morphology in the bluehead wrasse, Thalassoma bifasciatum (Bloch) (Teleostei: Labridae)

The effects of exogenous testosterone pellet implants at 21 and 40 days on the ovary of Thalassoma bifasciatum , a protogynous marine fish, are described. The characteristic markings of the terminal blue phase appeared by 4 to 5 days and were complete by 18 days. There was no shift in sex ratio in the treated fish when compared to an untreated control group. None of the ovaries showed signs of precocious transformation, i.e., spermatogenic tissue was lacking, and there was no evidence of duct formation. Instead, however, the ovaries of the treated fish showed marked degenerative changes characterized by oocyte breakdown, fat infiltration, vacuolization, the accumulation of fibrous PAS+ material and the appearance of small eosinophilic cells.
These findings conflict with earlier studies on the same organism that employed different procedures. However, they resemble more closely the results obtained from work on another protogynoid, the ricefield eel, Monopterus albus. They add new light on the role of sex hormones in the reversal process of T. bifasciatum and hermaphroditic fishes in general.     

Components of fertilization success in the bluehead wrasse, Thalassoma bifasciatum

In many species of marine organisms, males and females releasegametes directly into the water column. Although free-spawningmarine invertebrates appear to have highly variable fertilizationsuccess, in tropical reef fishes the average fertilizationsuccess is quite high, typically over 90%; nevertheless, substantialvariation has been reported, and fertilization has a directeffect on fitness. We investigated the factors affecting fertilization success in natural spawnings of the bluehead wrasse, Thalassoma bifasciatum. During a two-year study at a site in St. Croix,we found extensive and predictable variation in fertilizationsuccess in pair spawns of this reef fish. Fertilization successaveraged 95%, but was affected by the amount of sperm released,the water velocity at a site, the mating success of the male,and the size of the female. As sperm released in a spawn increases, and as water velocity at a site decreases, sperm concentrationsshould remain higher in the vicinity of eggs for a longer periodof time, and both of these factors are correlated with increasingfertilization success. The recent history of individuals withpartners or sites did not affect the fertilization successof their spawn. In an evolutionary context, the real and predictable variance in fertilization success in this species may influencethe mating choices of males and females. However, there iscurrently no evidence that females use differences in fertilizationsuccess among males or sites in their reproductive decisions.     

A multi-locus assessment of connectivity and historical demography in the bluehead wrasse (Thalassoma bifasciatum)

The pelagic larval stage of most coral reef fishes might allow extensive dispersal or, alternatively, some level of local recruitment might be important. Molecular markers can be used to obtain indirect estimates of dispersal to evaluate these alternatives, yet the extent of gene flow among populations is known for only a small number of species. The use of such markers must take into account the properties of the markers and the demographic history of the population when making inferences about current gene flow. In the Caribbean bluehead wrasse, Thalassoma bifasciatum, previous studies have found both substantial levels of local recruitment, in studies interpreting otolith microchemistry and, conversely, a lack of genetic differentiation inferred from mitochondrial DNA (mtDNA) restriction-fragment length polymorphism (RFLP) data and allozymes. However, if subtle differentiation exists, larger sample sizes and highly variable markers may be required to discern it. Here we present results from mitochondrial control region sequence and microsatellite data that indicate a lack of genetic differentiation at both small and large spatial scales. However, historical processes, such as changes in population size, may have affected the current distribution of genetic variation.     

Large mating aggregations and daily long-distance spawning migrations in the bluehead wrasse,Thalassoma bifasciatum

Synopsis The mating system of the labrid fishThalassoma bifasciatum has been extensively described in the literature, and the species has formed the basis for much work in ecology and behavior. However, most work in the past on this species has concentrated in small patch reef habitats. Here I describe a different mating pattern in bluehead wrasses occurring on a large, linear barrier reef in St. Croix, U.S. Virgin Islands. On this reef, very large group-mating aggregations (> 10000 fish) form daily at a single area on the forepart of the reef. Tagging studies showed that fishes are faithful to particular feeding schools arrayed along the forereef, and that they migrate to the spawning grounds along a specific path from distances more than 1.5 km away. No mating appears to occur in other upcurrent areas of the forereef. One-way migration times averaged 52 min from the farthest site studied, and feeding does not occur en route. Despite large differences in time spent in migration, no significant differences existed in fecundity or frequency of spawning among females living at different distances from the mating aggregation. The only apparent differences was a significantlyhigher growth rate in the fishes residing farthest from the aggregation. This higher growth rate corresponded to a higher overall feeding rate in the distant location, suggesting that food intake can more than compensate for the increased costs of migration.     

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

Synopsis 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-17. 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-17 production than did males. Production of both these steroids increased in a dose-related fashion with increasing doses of gonadotropin.     

Chromosomal analysis of repeated DNAs in the rainbow wrasse Coris julis (Pisces, Labridae)

Despite the interest of several authors, the karyotype of the labrid C. julis is still debated and in particular the presence of sex-chromosomes is still contradictory. In order to analyze the karyotype organization of C. julis we have performed an analysis with classical and molecular cytogenetic techniques. Our results after silver-, CMA₃- and DAPI-staining, C-banding and fluorescent in situ hybridization (FISH) using 28S rDNA, 5S rDNA, and telomeric repeat (TTAGGG) _n as probes allowed us to characterize the chromosomal location of several repetitive DNAs of C. julis. Finally, regardless of the technique used, no difference in the chromosome complement was found between males and females. This revised version was published online in August 2006 with corrections to the Cover Date.     

Benefits of heterospecific foraging by the Caribbean wrasse,Halichoeres garnoti (Pisces: Labridae)

Synopsis Terminal-phase yellowhead wrasses, Halichoeres garnoti, foraged alone or in association with foraging goatfish, Pseudupeneus maculatus and Mulloides martinicus. Whereas H. garnoti did not dig for benthic infauna, the goatfish foraged almost exclusively on and in sand substrata. Wrasses in the company of goatfish made significantly more strikes on sand substrata than did solitary wrasses, although there were no significant differences in the frequency of strikes on sand and hard substrata combined. The frequencies of other behaviors (searches, scratches, flights, chases, rests, yawns, and cleanings) were not significantly different between solitary and associated wrasses. H. garnoti are attracted to foraging goatfish, which provide access to an otherwise unavailable food resource. To whom reprint requests should be sent     

Terminal phase males stimulate ovarian function and inhibit sex change in the protogynous wrasse Thalassoma duperrey

Under experimental conditions, the probability of sex change in the protogynous wrasse Thalassoma duperrey is determined largely by an individual's relative size within a social group. Natural populations, however, contain two distinct male phenotypes that may also play a role in regulating sex change. To investigate potential effects of male phenotype, the ability to change sex, ovarian histology and serum estradiol-17 beta levels were examined in females maintained under controlled social settings. Large females housed with smaller or larger terminal phase males had significantly larger gonadosomatic indices than females housed singly, with other females or with smaller initial phase males. Similarly, ovaries of females housed with terminal phase males showed no histological evidence of sex change, whereas large females from other social groupings were in advanced stages of sex change. These results demonstrate terminal phase males inhibit sex change regardless of their size relative to the female. Furthermore, gonadosomatic indices, ovarian histology, and serum estradiol-17 beta levels of females housed with terminal phase males indicate normal ovarian function whereas ovaries of other treatment groups appear quiescent or are undergoing sex change. Consequently, terminal phase males may be required for normal ovarian development which may, in turn, inhibit sex change in T. duperrey.     

Spatial Dispersion and Nonmigratory Spawning in the Bluehead Wrasse (Thalassoma bifasciatum)

Earlier studies of the behavior of the bluehead wrasse Thalassoma bifasciatum have shown it to be a migratory spawner with large terminal-phase males defending temporary spawning territories. We describe a variant social structure where fish occupy permanent home ranges, spawn within or near these home ranges, and are not territorial. Movements of identified terminal-phase and intermediate males and females were mapped in the backreef areas of three coral reefs in Puerto Rico. Locations of spawns were then compared with these home-range maps. All fish used most or all of their morning home range during the afternoon spawning period. Both terminal-phase males and females spawned within or near their home ranges. Males were relatively tolerant of other males during the afternoon spawning period: only one in five encounters between large males resulted in aggressive chases. The location of chases bore no relation to spawning sites, areas of morning foraging, or to home-range borders. A Monte Carlo computer simulation was used to measure home-range dispersion of terminal-phase and intermediate males at our main site. If males were territorial, we should expect their home ranges to be significantly overdispersed within the cumulative area they occupied. During the first year male home ranges were overdispersed but during the second year they were randomly dispersed, even though the same number of terminal-phase males occupied the same study site both years. Differences in social behavior between backreef areas and other areas reported in the literature, and found in other portions of our reefs, may be related to differences in feeding ecology between populations and between different portions of the same reef.     

Social control of terminal phase transition in primary males of the diandric wrasse, <Emphasis Type="Italic">Halichoeres poecilopterus</Emphasis> (Pisces: Labridae)

In many diandric fishes, large territorial males with bright body coloration (terminal phase (TP) males) are derived either from initial phase (IP) females that change sex to male or from IP primary males that change color and behavior, but do not change sex. The mechanism controlling the transition of IP primary males into TP males is not well understood. We conducted cohabitation experiments to examine social conditions favoring TP transition by primary males in the diandric wrasse, Halichoeres poecilopterus. IP primary males always started TP-specific sexual behavior in the presence of a smaller subordinate, and subsequently acquired TP body coloration. In contrast, primary males under subordinate conditions often performed female-like sexual behavior. In pairs with similar body sizes, both individuals initiated TP male behavior. The results suggest that TP transition in primary males may be closely related to a dominance relationship (or size order) within social groups, as it is in the case of sex change by females.     

Karyological studies inCoris julis (Pisces,Labridae)

In the present investigation the diploid number 2n=48 (NF=58) has been determined for females, primary males, and secondary males ofCoris julis from the Gulf of Palermo. Differentiated sex chromosomes have not been observed in the population under study.     

sGnRH-A [(d-Arg6, Pro9, NEt-)LHRH] in combination with domperidone induces gonad reversal in a protogynous fish, the bluehead wrasse, Thalassoma bifasciatum

The effects of thrice weekly injections of Ovaprim (sGnRH-A in combination with domperidone) on the ovary of Thalassoma bifasciatum , a protogynous marine fish, after 4 and 6 weeks are described. At the end of 4 weeks of treatment, 72% ( n =13) of the original females showed signs of reversal. The majority of these were deemed early stage although mid- ( n =2) and late ( n =1) stage reversals were present. At 6 weeks, 92% of the females had ovaries that were undergoing transformation. None of these, however, progressed beyond the early stage. Only 3.8% of the control fish showed signs of reversal over the same periods. The blue head colour pattern, typical of the terminal phase, did not appear in any of the fish.
To our knowledge, this is the first report investigating the hypothalamic influence on sex reversal in T. bifasciatum . These observations shed new light on the neuroendocrine control of reversal of gender in this species and hermaphroditic fishes in general.     

Sex differentiation and early gonadal development in Bombina orientalis (anura: Discoglossidae)

Gonadal differentiation in premetamorphic Bombina orientalis is described and staged. The pattern of events during differentiation in Bombina differs in several respects from that previously described in other anurans. The Bombina gonad initially develops on the ventral surface of the vena cava, where there is no pre-existent somatic genital ridge prior to the arrival of the germ cells. The sexually undifferentiated gonad does not have a distinct cortex and medulla; instead, medullary cells ingress from the mesonephric blastema during sexual differentiation. Formation of a testis or an ovary appears to depend on the ability (or lack of ability) of the medulla to invade the germ cell-containing cortex. In the germ line, sexual differetiation can be recognized by a premeiotic increase in oogonial cell volume.     

Sex-inversion of the hermaphroditic, protogynous teleost Coris julis L. (Labridae)

Sex-inversion of Coris julis , a protogynous hermaphrodite, was studied using histological and cytological criteria. Four stages were recognized: oocyte and oogonial atresia; occurrence of spermatogonia with primordial germ cells (PGC) in the ovarian wall; onset of spermatogonial proliferation; considerable proliferation of spermatogonia and PGC with build-up of seminiferous tubules. Spermatogonia arise from PGC, undifferentiated and bipotential cells, in which mitotic activity was detected.