Organizational–Activational Concept Revisited: Sexual Differentiation in an Atherinomorph Teleost

Because of its numerous sexually dimorphic characters, the Western mosquitofishGambusia affinis affinisis an excellent vertebrate system for addressing questions concerning sexual differentiation. In this review, the actions and limits of gonadal sex steroids, specifically testosterone, on the development of the sexually dimorphic anal fin and its axial and appendicular support are described. Data from our laboratory show that the critical period in this species for the anterior transposition of the male anal fin and its appendicular support occurs during the late embryonic period and that this differentiation of the male phenotypic genital areaduring the critical period is regulated by androgen.     

Sexual Dimorphisms in the Dermal Denticles of the Lesser-Spotted Catshark,Scyliorhinus canicula (Linnaeus, 1758)

The dermal layers of several elasmobranch species have been shown to be sexually dimorphic. Generally, when this occurs the females have thicker dermal layers compared to those of males. This sexual dimorphism has been suggested to occur as a response to male biting during mating. Although male biting as a copulatory behaviour in Scyliorhinus canicula has been widely speculated to occur, only relatively recently has this behaviour been observed. Male S. canicula use their mouths to bite the female’s pectoral and caudal fins as part of their pre-copulatory behaviour and to grasp females during copulation. Previous work has shown that female S. canicula have a thicker epidermis compared to that of males. The structure of the dermal denticles in females may also differ from that of males in order to protect against male biting or to provide a greater degree of friction in order to allow the male more purchase. This study reveals that the length, width and density of the dermal denticles of mature male and female S. canicula are sexually dimorphic across the integument in areas where males have been observed to bite and wrap themselves around females (pectoral fin, area posterior to the pectoral fin, caudal fin, and pelvic girdle). No significant differences in the dermal denticle dimensions were found in other body areas examined (head, dorsal skin and caudal peduncle). Sexually dimorphic dermal denticles in mature S. canicula could be a response to male biting/wrapping as part of the copulatory process.     

The western mosquitofish (Gambusia affinis affinis): a new laboratory animal resource for the study of sexual dimorphism in neural circuits

The western mosquitofish (Gambusia affinis affinis) is a useful model for the study of sexual dimorphism and the neural circuits associated with sexual differentiation. This is largely because of its anal fin, which undergoes radical postnatal transformation in males. Understanding the neural mechanisms involved in this process may also help elucidate basic principles of the nervous system. The authors describe the mosquitofish as a model for research and present guidelines for the care and use of this species.     

Androgen dependent development of a modified anal fin, gonopodium, as a model to understand the mechanism of secondary sexual character expression in vertebrates

Male external genitalia show structural variations among species. Androgenic hormones are essential for the morphological specification of male type copulatory organs, while little is known about the developmental mechanisms of such secondary sexual characters. Western mosquitofish Gambusia affinis may offer a clue to the sexual differentiation researches, because they show a prominent masculine sexual character for appendage development, anal fin to gonopodium (GP) transition, and its formation could be induced in early juvenile fry by exogenously supplied androgens. We show that GP development is promoted by androgen dependent augmentation of sonic hedgehog (Shh) expression. Two AR cDNAs were cloned and identified as ARalpha and ARbeta from western mosquitofish. Both ARs were predominantly expressed in the distal region of outgrowing anal fin rays. Exposure of fry to androgen caused anal fin outgrowth concomitant with the Shh induction in the distal anal fin ray epithelium. When AR signaling was inhibited by its antagonist flutamide in fry, the initial induction of the Shh was suppressed accompanying retarded anal fin outgrowth. Similar suppression of anal fin outgrowth was induced by treatment with cyclopamine, an inhibitor of Shh signaling. These observations indicate that androgen dependent Shh expression is required for anal fin outgrowth leading to the formation of a genital appendage, the GP in teleost fishes. Androgen-induced GP formation may provide insights into the expression mechanism regulating the specification of sexual features in vertebrates.     

Sexually selected traits are larger and more variable in male than female beach-spawning capelin,Mallotus villosus

We evaluated whether morphological traits in capelin, Mallotus villosus, that appear to be sexually selected (pectoral fin, pelvic fin, anal fin, lateral ridge) were larger and more variable in males than females compared with naturally selected morphological traits (eyes, dorsal fin). Photographs were obtained of 136 capelin captured at two spawning sites and standardised measurements were taken of six morphological traits. Males had larger traits than females for a given body size and this was most pronounced in the traits thought to be sexually selected. Body size explained much of the variation in female traits but less variation in male traits, suggesting alternative selection pressures are involved. We suggest that larger male body size aids in endurance rivalry and sexually dimorphic traits help males to remain in physical contact with females while spawning on the beach.     

Permanent and Seasonal Expressions of Sexual Dimorphisms in a Weakly Electric Fish, Mormyrus rume proboscirostris Boulenger 1898 (Mormyridae, Teleostei)

Several sexually dimorphic characters of the anal-fin complex in the mormyrid fish, Mormyrus rume proboscirostris, assist during courtship when the male envelops the female's anal fin with its own to form a common spawning pouch (anal-fin reflex). We found that developmental growth and seasonally cycling gonadal activity selectively affect their expression. The structures defining the anal fin undergo a permanent sexually dimorphic transformation at a time when ripe spermatozoa first appear in the testis of young males. However, the expression of a dorsally directed indentation of the posterior ventral body wall, affecting the dorsal margin of the anal fin, appeared to be more plastic as it correlated with the gonadosomatic index, that is, testis size. We surmised that this indentation is influenced by cyclic anabolic action on muscle involved with the execution of the anal-fin reflex.     

摄食水平对不同性别食蚊鱼仔幼鱼生长发育的影响

在实验室27℃水温下,研究了少食、中食和饱食三个摄食水平对0至25日龄雌雄食蚊鱼(Gambusia affinis)的生长发育特征和饵料利用效率的影响。26d饲养实验结束后,对试验鱼摄食和生长指标、臀鳍分化、性成熟及饵料转换效率进行分析。结果显示:到臀鳍开始分化时,饱食组雄鱼的累计摄食总能量和生长速度开始小于雌鱼,且随日龄的增加差异加大;随摄食水平的增加,0日龄仔鱼到臀鳍分化和性成熟的时间缩短。至实验结束,各摄食组的雄鱼均形成发育完善的生殖足,性腺都达到成熟状态;而雌鱼性成熟迟于雄鱼,且其性成熟更易受到摄食水平的影响,饱食组只有约50%的个体达到性成熟,少食组的卵母细胞则均处在小生长期。随着摄食水平的增加,雌雄鱼的体长、体重和干物质特定生长率均呈明显上升趋势,而干物质饵料转化效率则呈明显下降趋势;实验结束时,雌鱼的生长指标和干物质饵料转化率均大于雄鱼。以上结果表明,伴随臀鳍的分化,食蚊鱼在摄食、生长、发育、性成熟和应对食物丰度变化上表现出显著的性别差异。       

Development of secondary sexual characters and their relationship to ontogeny and seasonal reproductive period in Hyphessobrycon igneus (Ostariophysi: Characiformes)

Sexual dimorphism in size, anal‐fin shape and coloration of Hyphessobrycon igneus, Characidae, were examined. Males were more frequent at larger body sizes, confirming body size as a sexually dimorphic trait. Anal‐fin shape and the colour of all fins were the same for females and juveniles, differing only in adult males. Likewise, only adult males had bony hooks on fin rays; larger and more sexually mature males had the most numerous and developed hooks and hooks were most developed in degree and number during peak reproductive periods. Fin hooks regressed in number and developmental degree after the reproductive period, but restarted development with the beginning of the new reproductive period without completely disappearing. Results show that bony hooks have a development and regression cycle related to reproductive seasonality.     

Sexual differentiation of brain and behavior in quail and zebra finches: Studies with a new aromatase inhibitor, R76713

In many species of vertebrates, major sex differences affect reproductive behavior and endocrinology. Most of these differences do not result from a direct genomic action but develop following early exposure to a sexually differentiated endocrine milieu. In rodents, the female reproductive phenotype mostly develops in the absence of early steroid influence and male differentiation is imposed by the early action of testosterone, acting at least in part through its central conversion into estrogens or aromatization. This pattern of differentiation does not seem to be applicable to avian species. In Japanese quail (Coturnix japonica), injection of estrogens into male embryos causes a permanent loss of the capacity to display male-type copulatory behavior when exposed to testosterone in adulthood. Based on this experimental result, it was proposed that the male reproductive phenotype is “neutral” in birds (i.e. develops in the absence of endocrine influence) and that endogenous estradiol secreted by the ovary of the female embryo is responsible for the physiological demasculinization of females. This model could be recently confirmed. Females indeed display a higher level of circulating estrogens that males during the second part of their embryonic life. In addition, treatment of female embryos with the potent aromatase inhibitor, R76713 or racemic vorozole™ which suppresses the endogenous secretion of estrogens maintains in females the capacity to display the full range of male copulatory behaviors. The brain mechanisms that control this sexually differentiated behavior have not been identified so far but recent data suggest that they should primarily concern a sub-population of aromatase-immunoreactive neurons located in the lateral parts of the sexually dimorphic preoptic nucleus. The zebra finch (Taeniopygia guttata) exhibits a more complex, still partly unexplained, differentiation pattern. In this species, early treatment with exogenous estrogens produces a masculinization of singing behavior in females and a demasculinization of copulatory behavior in males. Since normal untreated males sing and copulate, while females never show these behaviors even when treated with testosterone, it is difficult to understand under which endocrine conditions these behaviors differentiate. In an attempt to resolve this paradox, we recently treated young zebra finches with R76713 in order to inhibit their endogenous estrogens secretion during ontogeny and we subsequently tested their behavior in adulthood. As expected, the aromatase inhibitor decreased the singing frequency in treated males but it did not affect the male-type copulatory behavior in females nor in males. In addition, the sexuality differentiated brain song control nuclei which are also masculinized in females by early treatment with estrogens, were not affected in either sex by the aromatase inhibitor. In conclusion, available data clearly show that sexual differentiation of reproductive behaviors in birds follows a pattern that is almost opposite to that of mammals. This difference may be related to the different mechanisms of sex determination in the two taxa. In quail, the ontogeny of behavioral differentiation is now well understood but we only have a very crude notion of the brain structures that are concerned. By contrast, in zebra finches, the brain mechanisms controlling the sexually differentiated singing behavior in adulthood have been well identified but we do not understand how these structures become sexually dimorphic during ontogeny.     

The effects of induced masculinization on reproductive and aggressive behaviors of the female mosquitofish,Gambusia affinis affinis

Synopsis FemaleGambusia affinis affinis were masculinized with the degraded products of 65% stigmastanol-30% B-sitosterol, a phytosterol. The masculinized females were paired with non-treated males, non-treated females, and other masculinized females. The pairs were analyzed for reproductive and aggressive behaviors exhibited. The behavioral patterns of these pairs were compared statistically to the behavior patterns of the following non-treated pairs: male with male, female with female, and male with female. The masculinized females behaved like males in that they followed, swung, and thrust their gonopodia at non-treated females and larger masculinized females. However, the male-like behavior of the masculinized females was not as intense as that of normal males. Also, the effect of masculinization on the behavior of the treated female was context-dependent as seen when placed with a male or a smaller masculinized female. Under these conditions the masculinized females behaved like typical females and exhibited no masculinized behaviors. The effects of anal spots and size differences are discussed as possible explanations for the variability in behaviors exhibited. The masculinized females displayed no change in aggressive behaviors.     

Differential regulation of msx genes in the development of the gonopodium,an intromittent organ,and of the "sword," a sexually selected trait of swordtail fishes (Xiphophorus)

The possession of a conspicuous extension of colored ventral rays of the caudal fin in male fish of swordtails (genus Xiphophorus) is a prominent example for a trait that evolved by sexual selection. To understand the evolutionary history of this so-called sword molecularly, it is of interest to unravel the developmental pathways responsible for extended growth of sword rays during development of swordtail males. We isolated two msx genes and showed that they are differentially regulated during sword outgrowth. During sword growth in juvenile males, as well as during testosterone-induced sword development and fin ray regeneration in the sword after amputation, expression of msxC is markedly up-regulated in the sword forming fin rays. In contrast, msxE/1 is not differentially expressed in ventral and dorsal male fin rays, suggesting a link between the development of male secondary sexual characters in fins and up-regulation of msxC expression. In addition, we showed that msx gene expression patterns differ significantly between Xiphophorus and zebrafish. We also included in our study the gonopodium, a testosterone-dependent anal fin modification that serves as a fertilization organ in males of live-bearing fishes. Our finding that increased levels of msxC expression are associated with the testosterone-induced outgrowth of the gonopodium might suggest either that at least parts of the signaling pathways that pattern the evolutionary older gonopodium have been coopted to evolve a sexually selected innovation such as the sword or that increased msxC expression may be inherent to the growth process of long fin rays in general.     

Sexual behaviour of immature male eastern mosquitofish: a way to measure intensity of intra-sexual selection?

Immature males of eastern mosquitofish Gambusia holbrooki start to be sexually active well before their copulatory organ (gonopodium) has completely developed and before they become able to transfer sperm. Sexual activity of males, consisting of copulatory attempts tending to bypass female acceptance, is intense (one attempt per minute) and is likely to be energetically very costly. The sexual behaviour of immature males relative to their maturation stage is described and tested against two possible adaptive explanations. Sexual activity was present in males from the beginning of the development of their gonopodium and increased during the following stages of maturation. Two to three weeks before gonopodium development was completed, sexual activity of immatures was as high as that of adults. Adult males showed aggressive behaviour against a male attempting a copulation, irrespective of the maturity of the latter. Since previous studies have shown that the reproductive success in this species is negatively correlated with male size when male–male competition is low (i.e. when the sex ratio is female biased), but decreases with male size when competition is high, the hypothesis was tested that sexual activity of immature males functions as a way to predict their future reproductive success if they mature at a given size. A second hypothesis tested was that precocious sexual experience improves the efficiency of copulatory attempts. Results were more in agreement with the first hypothesis, since size at maturity of males was influenced by the sex ratio experienced during maturation and precocious experience gave very little advantage.     

Normally occurring intersexuality and testosterone induced plasticity in the copulatory system of adult leopard geckos

The copulatory neuromuscular system of lizards is highly sexually dimorphic. Adult males possess bilateral penises called hemipenes, which are independently controlled by two muscles, the retractor penis magnus (RPM) and transversus penis (TPN). These structures are not obvious in adult females. However, in adult female leopard geckos (Eublepharis macularius), testosterone induces hemipene growth. We investigated whether these structures develop de novo in adulthood or are histologically present as rudimentary structures in the female leopard gecko. We also investigated the extent of sexual dimorphisms and plasticity in the associated neuromuscular components. To do this, we compared copulatory morphology (sizes of hemipenes, RPM and TPN muscle fibers, and associated motoneurons, as well as motoneuron and RPM fiber number) in adult females treated with testosterone, control females, and control males. All of the geckos possessed hemipenes, RPMs and TPNs, but these structures were indeed vestigial in control females. Testosterone induced striking increases in hemipene and copulatory muscle fiber size in females, but not to levels equivalent to control males. In parallel, males with increased levels of androgenic activity had larger hemipenes, suggesting naturally occurring steroid-induced plasticity. Copulatory motoneurons were not sexually dimorphic in size or number, and these measures did not respond to testosterone. The data demonstrate that the copulatory system of leopard geckos, in which gonadal sex is determined by egg incubation temperature, differs from that of many species (both reptilian and mammalian) with genotypic sex determination. Indeed, the system is remarkable in that adult females have normally occurring intersex characteristics and they exhibit substantial steroid-induced morphological plasticity in adulthood.     

Experimental manipulation reveals a trade‐off between weapons and testes

Theory predicts a trade‐off between sexually selected weapons used to secure mates and post‐copulatory traits used to maximize fertilization success. However, individuals that have a greater capacity to acquire resources from the environment may invest more in both pre‐ and post‐copulatory traits, and trade‐offs may not be readily apparent. Here, we manipulate the phenotype of developing individuals to examine allocation trade‐offs between weapons and testes in Mictis profana (Hemiptera: Coreidae), a species where the hind legs are sexually selected weapons used in contests over access to females. We experimentally prevented males from developing weapons by inducing them to autotomize their hind legs before the final moult to adulthood. We compared trait expression in this group to males where autotomy was induced in the mid‐legs, which are presumably not under sexual selection to the same extent. We found males without weapons invested proportionally more in testes mass than those with their mid‐legs removed. Males that developed to adulthood without weapons did not differ from the mid‐leg removal group in other traits potentially under precopulatory sexual selection, other post‐copulatory traits or naturally selected traits. In addition, a sample of adult males from the same population in the wild revealed a positive correlation between investment in testes and weapons. Our study presents a critical contribution to a growing body of literature suggesting the allocation of resources to pre‐ and post‐copulatory sexual traits is influenced by a resource allocation trade‐off and that this trade‐off may only be revealed with experimental manipulation.     

Sexual dimorphism and allometry of external morphology in Oreochromis mossambicus

Sexual dimorphism in growth of conventional morphometric characters was investigated in juveniles and young adults (size range: 31 to 91 mm) of Oreochromis mossambicus . A closely associated set of traits was identified that shows sexually dimorphic growth, which was positively allometric in the males. These traits correspond to two different morphological complexes: jaw structure and anal/dorsal fins. The best sex discriminators among this set of traits were premaxilla width, anal fin height and snout length. These findings may be explained in terms of intra– and inter–sexual selection acting together and favouring males with strong and large mouths and high dorsal and anal fins, traits that are important in agonistic displays (jaw and fins), fighting and nest digging (jaw).     

New remarkable sexually dimorphic miniature species of Hyphessobrycon (Characiformes: Characidae) from the upper Rio Tapajós basin

A new miniature species of Hyphessobrycon is described from an affluent of the Rio Papagaio, tributary of the Rio Juruena, upper Rio Tapajós basin, Mato Grosso State, Brazil. The new species can be distinguished from its congeners by having a remarkable secondary sexual dimorphism in its live colouration (males are red and females yellow), well‐defined and relatively narrow dark midlateral stripe extending from tip of snout to tip of middle caudal‐fin rays, absence of humeral blotch, 15–18 branched anal‐fin rays and five or six branched pelvic‐fin rays. The sexually dimorphic colouration of the new species is briefly discussed regarding its temporal variation.     

Effects of 17α-Methyltestosterone on Sexually Dimorphic Characters in the Weakly Discharging Electric Fish,Brienomyrus niger(Günther, 1866) (Mormyridae): Electric Organ Discharge, Ventral Body Wall Indentation, and Anal-Fin Ray Bone Expansion

Adult males of African weakly discharging electric fish (family: Mormyridae) are distinguished from juveniles and adult females by a dorsally directed indentation of the posterior ventral body wall and by massive bone expansion of the bases of a select number of anal-fin rays. These sexually dimorphic structures seem to facilitate the anal-fin reflex that is displayed during courtship when the male envelopes its anal fin around the female's to form a common spawning pouch. Expanded bone could provide additional surface for muscle attachment and thus assist in part with the courtship sequence. Based on the fact that the expression of the male sexually dimorphic electric organ discharge (EOD) is under androgen control, and that the female EOD can be masculinized through testosterone administration, we hypothesized that androgens should also drive anal-fin ray bone expansion in male mormyrids and equally effect male-like changes in treated juveniles and adult females. Exogenous androgen treatment (17α-methyltestosterone) of adult femaleBrienomyrus nigerresulted in a male-like EOD, and male-typical structural transformations (body wall indentation and anal-fin ray bone expansion). Some of these changes were immediate and receded following hormone withdrawal (EOD), while others developed more slowly and were apparently permanent (indentation and bone formation). 17α-Methyltestosterone administration affected only those targets in females that are normally involved in the male's reproductive behavior, i.e., its courtship signal (EOD) and two morphological features (body-wall indentation and bone expansion). Rays of the dorsal or caudal fins were never affected.     

Female masculinization and reproductive life history in the western mosquitofish (<Emphasis Type="Italic">Gambusia affinis</Emphasis>)

In this study, we explored the relationship between female masculinization and various morphological and reproductive parameters of male and female western mosquitofish (Gambusia affinis), including body size, anal fin length, testes mass and female fecundity. We detected high levels of female masculinization in the Brazos and Little Brazos Rivers (Falls County, Texas) in March 2008, but not 2009. In both populations, masculinized females were smaller, possessed shorter anal fins, and had significantly lower total fecundity (collective number of eggs and embryos) compared to non-masculinized females. In addition, 2008 non-masculinized females were smaller and less fecund than 2009 non-masculinized females in both populations. Moreover, 2008 males were smaller, had shorter gonopodia, and smaller testes than 2009 males. Our results support the hypothesis that female masculinization can have a strong influence on reproductive life history and population dynamics of livebearing fishes.     

利用食蚊鱼目标基因转录水平评价东莞寒溪河雌/雄激素物质污染现状

应用食蚊鱼(Gambusia affinis)的卵黄蛋白原(VTGa)基因和雌/雄激素受体(ERa/ARa)基因转录水平为指标, 评价广东东莞寒溪河受雌/雄激素物质污染的现状。结果显示, 与对照点从化流溪河(LX)相比, 寒溪河松山湖(SM)、杨屋村(YW)、横沥镇(HL)、樟村污水处理厂上游(ZU)和樟村污水处理厂下游(ZC)各采样点雄性成年食蚊鱼肝脏中的VTG转录水平都具有显著或极显著水平的升高(p0.05和p0.001), 分别为7.67、169.43、148.69、152.01和98.12倍; 而较之对照点, 雄鱼臀鳍的雌激素受体ER转录水平仅HL位点有显著性升高(p0.05), 其余位点则不明显(p0.05)。此外, 寒溪河SM、YW、HL、ZU和ZC各采样点雌鱼臀鳍雄激素受体AR转录水平分别是对照点从化流溪河(LX)的1.21、0.82、0.34、0.41和0.89倍, 但是只有HL和ZU点有显著性减小(p0.05), 其余各点的变化差异不显著(p0.05)。研究结果显示, 生活在东莞寒溪河中的食蚊鱼受雌/雄激素物质干扰明显, 普遍出现显著的雌激素效应, 表明寒溪河水体受雌激素物质污染更严重。此外证明, 鱼类肝脏VTGa基因相对于臀鳍ERa/ARa基因转录水平更适合作为监测水体环境雌/雄激素物质的生物标志物。       

Alternative male mating tactics in Gobius niger

In the field, parental males of the black goby Gobius niger were always >9 cm L _T, showed a conspicuous elongation of the first dorsal fin, and were darkly coloured. Larger parental males did not occupy larger nests or obtain more eggs, suggesting that over a threshold size reproductive success was not correlated with male size. The mating system was polygynous with different egg clutches simultaneously present in a nest. Smaller sexually mature males were observed around and inside nests in which spawning was occurring. These opportunistic males ranged in body size between 6 to 8 cm L _T. In contrast to parental males, they did not show an exaggeration of the first dorsal fin and were cryptically coloured, thus resembling small females in their external morphology. Aquarium experiments confirmed that smaller males perform a sneaking behaviour, releasing sperm when parental males spawn. As a consequence, it is possible to define three male types: type I are small, without an elongated dorsal fin and adopt a sneaking mating tactic; type III are large, have a pronounced elongation of the dorsal fin and perform parental behaviour; type II are intermediate both in size and elongation of the first dorsal fin and behave as either as sneakers or, possibly, as parental males according to nest availability and male competition.