Morphology, endocrinology, and environmental modulation of gonadal sex differentiation in teleost fishes

Successful reproduction by an adult depends on the normal ontogenesis of the gonads, a complex process of cellular and histological differentiation that starts early in life. This process is theoretically predetermined by genetic factors and includes sensitisation of the bipotential gonads to endogenous endocrine factors prior to, during and even after commitment to maleness or femaleness. However, young fish are relatively vulnerable to a host of environmental (physical and chemical) factors that can affect this endogenous endocrine axis, disturbing or even overriding the putative developmental pathway. This sexually lability can be exploited to our advantage for the production of monosex fish populations of the most valuable sex for food production or aquarium fish trade. On the other hand, it represents also a potential path for undesirable influences from endocrine-disrupting chemicals and climatic factors, particularly environmental temperature. This paper provides a detailed account of the early histological process of gonadal sex differentiation, with special reference to gonochoristic species, and reviews the criteria employed to positively identify ovarian and testicular differentiation. It also reviews the development of endocrine competence and sensitivity of the differentiating gonads to exogenous influences in the context of the relative stability of genotypic sex determination in various fish species. Sex differentiation in some species seems to be under strong genetic control and may not require endogenous sex steroid production. Conversely, reliance on endogenous sex steroids for gonadal differentiation is observed in other species and this phenomenon is apparently associated with a higher incidence of environment (mainly temperature)-labile sex differentiation. This revised version was published online in July 2006 with corrections to the Cover Date.     

采用RAPD技术筛选马口鱼性别差异相关分子标记

选用SBSA和SBSB两组随机引物（各20条）分别对马口鱼卵巢DNA和精巢DNA进行RAPD扩增,并用1．5％的琼脂糖凝胶电泳检测扩增结果。结果表明,在40条随机引物中,有11条引物可以扩增出明显的雌雄性别差异条带,条带范围以SBSA5、SBSA6、SBSA9、SBSA16、SBSA19、SBSA20、SBSB2、SBSB3、SBSB9、SBSB16、SBSB17为主。运用RAPD筛选获得马口鱼性别DNA标记将有助于了解其调控雌雄性别发生的差异基因,为进一步的遗传育种提供基础。     

Gonadal morphogenesis and sex differentiation in cultured chub mackerel,Scomber japonicus

The timing of primordial germ‐cell (PGC) migration with regard to the gonadal anlagen, gonad formation and sex differentiation was examined histologically in the chub mackerel (Scomber japonicus) at 5–190 days post hatching (dph). At 5 dph, PGCs appeared on the peritoneal epithelium surface or in the mesentery, on the dorsal side of the abdominal cavity. By 10 dph, stromal cells around the PGCs proliferated. The gonadal primordium was formed by 15 dph. The gonadosomatic index was 0.01% at 30 dph and increased thereafter (0.32% in females and 0.04% in males at 160 dph). Ovarian differentiation occurred at 30–40 dph, indicated by ovarian cavity formation (elongation and fusion of the upper and lower ovarian edges). Meiosis was subsequently initiated. A few meiotic oocytes surrounded the cavity at 50 dph; most were in the perinucleolus stage at 60 dph and attained a diameter of 60–70 μm at 190 dph. Testicular differentiation occurred at 30 dph, indicated by the formation of the sperm duct primordium. Spermatogonia gradually proliferated, developing into spermatocytes at the chromatin–nucleolus stage (after 90 dph) and subsequently into spermatids and spermatozoa (160 dph). These data could aid the development of seeding and cell‐engineering technologies for scombrid fish.     

Effects of a nonsteroidal aromatase inhibitor on gonadal differentiation of bluegill sunfish Lepomis macrochirus

In the present study, the efficacy of Letrozole, a potent nonsteroidal aromatase inhibitor (AI), on gonadal sex differentiation and sex reversal was examined in bluegill sunfish (Lepomis macrochirus). In Experiment 1, using AI diet treatments (50, 150, 250 and 500 mg kg⁻¹) from 30 to 90 days posthatch (dph), AI interrupted ovarian cavity formation at a dose of 500 mg kg¹ diet and one intersex fish was identified in this group. The proportions of males in all the treated groups were significantly higher than those in the control group. In Experiment 2, using AI immersion treatments (250, 500 and 1000 μg L⁻¹) during 30–50 dph, the treated groups of 500 and 1000 μg L⁻¹ produced significantly more males than the control and 250 μg L⁻¹ groups. Histological examination revealed no differences in ovary or testis tissue between control and AI‐treated fish. There were no significant differences detected in body weight and length among the AI treated and control groups (P>0.05) for both experiments. The results from these two experiments suggest that inhibition of aromatase activity by AI could influence sex differentiation in bluegill sunfish.     

Effects of estradiol-17β and 17α-methyltestosterone on gonadal sex differentiation in the F2 hybrid sturgeon, the bester

The effects of giving oral estradiol-17β (E₂) and 17α-methyltestosterone (MT) on gonadal sex differentiation in the F₂ hybrid sturgeon, the bester ( Huso huso female ×  Acipenser ruthenus male), are investigated. Giving E₂ at 10 μg/g diet to fish from 14 months until 31 months of age induced incomplete feminization and resulted in approximately 40% abnormal ovary development in which oocytes were observed without ovarian lamellar structures and gonadal shape was similar to normal testis. Giving MT at 25 μg/g diet for the same duration failed to induce masculinization, and resulted in approximately 30% undeveloped gonads even at 30–37 months of age. In contrast, E₂ and MT at only 1 μg/g diet given from 3 to 18 months of age was sufficient to induce feminization and masculinization, respectively. In these fish, feminization and masculinization were observed at 9 months, when most putative ovaries and testes were histologically distinguishable by the shape of the gonadal surface. These results indicate that sex reversal can be induced in these fish by hormone treatment that is started at 3 months age, before morphological differentiation occurs on the stroma of the gonads.     

Potential use of otolith microchemistry for field studies of temperature-dependent sex determination and gonadal degeneration in fish

This paper describes a novel approach to study the expression of temperature-related reproductive phenomena such as thermolabile sex determination (TSD) and heat-induced germ cell loss in wild fish populations. The proposed approach is based on the reconstruction of the past thermal history of individual fish through the microchemical analysis of the otoliths (mineralized structures responsible for the sense of balance). As an example, we outline the preliminary results of an investigation on the natural occurrence of TSD in pejerrey Odontesthes bonariensis, an atherinid fish in which all-female to all-male populations can be produced by experimental manipulation of the rearing temperature during the critical time of sex differentiation, from Lake Kasumigaura, Japan.     

Patterns of sex ratio response to water temperature during sex determination in honmoroko Gnathopogon caerulescens

ABSTRACT:   The sex ratios of the offspring of males from broods of honmoroko Gnathopogon caerulescens that displayed thermolabile sex determination (TSD), and those from eggs collected from the wild, strongly suggest that the combination of parents plays an important supplementary role in TSD in this species. The proportion of females in all broods from eight pairings of fish captured in the field decreased significantly at 30°C compared to 20°C, and one brood was entirely female at the lower temperature. These results suggest that phenotypic males (XX-males) exist in nature, probably as a result of sex change from genetic females caused by TSD. Sex ratios in relation to water temperature fall into five patterns, two of which seem to result from normal pairings of XX-females and XY-males, and from pairings of XX-males and XX-females with non-thermosensitivity. Two other patterns are thought to result from the same types of pairing, but with thermosensitivity. The last pattern, which shows male bias at both low and high temperature, is not explained only by the combination of genotypic sex determination (XX/XY) and TSD. Fluctuating temperatures close to natural conditions showed little potential to masculinize broods laid in the field between April and early June. However, the sex ratios of fish spawned in early June showed male bias at low and high temperatures. The sex determination mechanism in G. caerulescens may involve the interaction between temperature and sex-determining genetic factors, and the relative importance of each component differs with breeding season.     

The role estrogens play in sex differentiation and sex changes of fish

Using genetically controlled all-female and all-male tilapia (Oreochromis niloticus), the role steroid hormones play in the sex differentiation was analyzed histologically, ultrastructurally, immunohistichemically and experimenntally. The results strongly suggest that endogenous estrogen acts as an ovarian inducer, and that the lack of steroid hormone including androgen is important for testicular differentiation. Moreover, the roles of steroid hormones in protogynous sex change of three-spotted wrasse (Halichoeres trimaculatus) and saddleback wrasse (Tharassoma duperrey) were examined. The results strongly support the hypothesis that the endogenous estrogen plays an important role in protogynous sex change.     

The effects of temperature on sex differentiation and growth of black sea bass (Centropristis striata L.)

To examine the effects of temperature on sex differentiation in the black sea bass ( Centropristis striata L.), a protogynous hermaphrodite, juveniles (∼0.5 g) were cultured in recirculating systems at 17, 21 or 25 °C. Growth was assessed at 155, 182, 241 and 275 days post hatch and sex differentiation was determined histologically. No differences were found in the sex ratios of fish reared at different temperatures, but only 55–64% developed as females. Growth was significantly greater in males across all temperature treatments. These results suggest that black sea bass exhibit sexually dimorphic growth patterns and that female-specific sex determination can be disrupted in culture.     

The influence of phytoestrogens and oestradiol-17β on growth and sex determination in the European eel (Anguilla anguilla)

Under aquaculture conditions, European eels (Anguilla anguilla) produce a high percentage of males (80–95%) that normally stop growing at 100–200 g. Females continue to grow to 500–750 g and obtain higher market value. Therefore, increasing the percentage of females in a population would be beneficial to the culture of eels. The present study was carried out in order to examine the effect of oestradiol and phytooestrogens on sex differentiation and growth rate of eels. Juvenile European eels with undifferentiated gonads were fed pellets containing oestradiol‐17β (E2) or phytooestrogens for 100–150 days. Feeding E2 resulted in 50–61% increase in body weight compared with the control. Oestradiol‐17β and phytooestrogens both elevated significantly the percentage of females in the population. Feeding E2 at 20 mg kg^?1 feed resulted in 70% females, while lower concentration of E2 (2 mg kg^?1) resulted in only 30% after 100 days (Experiment 1). The same dose given for 150 days (Experiment 2) resulted in 88% females, indicating that both, the concentration and duration of E2 treatments had a significant effect on sex differentiation. Fish fed genistein at 2 mg kg^?1 for 100 days, resulted in 55% of females, but at a higher dose of 20 mg kg^?1 there were only 15% females. These results demonstrate that phytooestrogens can be used as alternatives to gonadal steroids for sex manipulation in eels, but the optimal concentrations and duration are still to be determined.     

Application of sex reversal technology in ornamental fish culture

Many species of ornamental fish, includingboth egg-layers and livebearers, exhibit a marked sexualdimorphism due to the more pigmented bodies and larger finsusually observed in males. This results in male fish commandingup to four times the price of females. Because of this pricediscrepancy, the culture of monosex (all-male) stocks ofornamental fish could be of a significant economic advantage.This paper outlines the current sex reversal technology alreadydeveloped for several fish species important for aquaculture andexamines its potential applicability to ornamental fish culture.The methods to obtain all-male stocks include direct androgentreatment and the production of YY males. The direct approach iseasy to implement and straightforward. However, in somelivebearers such as the guppy, treatment of newly born fry iscomplicated because they already have sexually differentiatinggonads and the masculinization may be temporary. In this regard,the paper stresses the importance of correctly timing treatmentin relation to gonadal development to achieve permanentmasculinization     

Determination of sex and gonadal maturity in the honeycomb grouper, <Emphasis Type="Italic">Epinephelus merra</Emphasis>, through biopsy

Determination of the sex and gonadal maturity of fish is necessary for successful commercial aquaculture operations. Sex identification is problematic in grouper fish, because they change sex at certain ages or sizes. Here we represent an accurate and reliable method for sex determination of the live grouper, using 5–10 mg gonadal tissue samples, harvested using non-lethal gonadal biopsy. Sex and sexual maturity of biopsied individuals were determined using standard histological method. All biopsied fish survived and there were no serious infections resulting from the surgery. Therefore, this biopsy technique is a simple and cost-effective tool for grouper fishery management.     

A review of sex determination and searches for sex‐specific markers in sturgeon

The availability of monosex populations of caviar‐producing female sturgeon would considerably enhance the economic viability of domestic caviar production systems. However, it is not possible to distinguish males from females by morphological characters at larval, juvenile and even adult stages. The mechanism of sex determination in sturgeons is poorly understood, and to date no sex‐specific markers in sturgeon have been reported. This review concentrates on the methodologies used to elucidate the mode of sex determination in sturgeon species and provides information on the molecular tools used to determine genetic sex markers.     

两种液化蛋白及其在饲料中的应用

IntroductionFish silage has been produced and used as high nutrition protein resource in feed for many years in Nor-way.The main raw materials are fish offals from fish processing industry,dead fish from fish farming factoryand pelagic fish.It is now a successful and profitable industry and benefits environment protection.With therapid growth in fish farming in China,the demands for high quality protein resource are increasing.There isgreat potential for adopting silage technology in China.A…