Increase in estrogen signaling in the early brain of orange-spotted grouper Epinephelus coioides: a mini-review

Despite neurosteroidogenic enzymes are playing important roles in the regulation of brain development and function, the potential link between brain and gonad by the action of steroid hormones during gonadal sex differentiation is still not clear in teleosts. In this mini-review, we summarized our understanding on the early brain development related to the synthesis of neurosteroids and receptor signaling during gonadal sex differentiation in protogynous orange-spotted grouper, Epinephelus coioides (functional females for the first 6 years of life and start to sex change around the age of 7 years) and protandrous black porgy (functional males for the first 2 years of life but begin to change sex during the third year). We found a similar profile in the increased expression of brain aromatase gene (aromatatse B or cyp19a1b), aromatase activity, estradiol (E₂), and estrogen signaling in the brain of both grouper and black porgy fish during gonadal sex differentiation. In contrast to mammals, teleost fish Cyp19a1b expressed in a unique cell type, a radial glial cell, which is acted as progenitors in the brain of developing and adult fish. In agreement with these pioneer studies, we demonstrated that the grouper cyp19a1b/Cyp19a1b was expressed in radial glial cells. Further, in vivo data in the grouper brain showed that exogenous E₂ upregulated Cyp19a1b immunoreactivity (ir) in radial glial cells. These data suggest the possible roles of Cyp19a1b and E₂ in early brain development which is presumably related to gonadal sex differentiation.     

Expression of dmrt1 and sox9 during gonadal development in the Siberian sturgeon (Acipenser baerii)

The sex differentiation period of the Siberian sturgeon was investigated through expression profiling of two testicular markers (dmrt1 and sox9). At the molecular level, a clear sexual dimorphism of dmrt1 and sox9 was observed in 3-year-old fish with immature gonads, in which males showed higher expression of these genes. Among 16-month-old sturgeons cultured in Uruguay, gonad morphology analyses showed one group of fish with undifferentiated gonads and a second group which had started their histological differentiation into ovaries or testes. dmrt1 showed a significantly higher expression in testes of recently differentiated fish, but this was not the case for sox9. In undifferentiated fish, we observed two clearly different groups in terms of expression: one group of fish over-expressing male markers (dmrt1, sox9) and another group of fish showing very low expression of these genes. This suggests that fish undergoing male differentiation can be identified by their profiles of gene expression before they undergo morphological differentiation.     

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.     

Growth Rate, Maturation Level and Flesh Quality of Three Strains of Large Rainbow Trout (Oncorhynchus Mykiss) Reared in Estonia

Three all-female strains of large rainbow trout imported to Estonia (Danish, Finnish and Donaldson strain) were evaluated in a communal rearing experiment in tanks. The fish had been reared on different fish farms before the experiment started and they had an initial weight of 400–500 g. There were no significant genetic differences between the investigated strains on the basis of three allozyme loci. The total weight gain of the strains during the trial was similar but differences were revealed between seasons. The growth rate of the Finnish trout was high during the first autumn but decreased during the second summer of the trial, probably due to infestation by eye fluke Diplostomum sp. The strains differed in the percentage of immature fish and in the level of gonad development at the age of 2.5–3 years. The Donaldson strain had the highest number of immature fish of the studied strains. Mature females of the Finnish strain had the smallest gonads but also the lowest percentage of immature fish. The level of maturation was the main factor determining the quality – slaughter yield and flesh color.     

Gonadogenesis in scallop Chlamys farreri and Cf‐foxl2 expression pattern during gonadal sex differentiation

Gonad development and sexual differentiation are important biological processes. More insight is needed in these processes to improve selection and breeding efficiency in the production of culture animals for monosex farm operations of commercially important shellfish species. Foxl2 is the earliest known sex dimorphic marker of ovarian differentiation in mammals, but studies on foxl2 in invertebrate species are rare. This study revealed the gonadogenesis and expression characteristics of foxl2 during the gonadal differentiation in the scallop Chlamys farreri, an important commercial mollusk in China. The result showed that C. farreri gonad becomes visible once juveniles reach a shell height of 5.5 mm. Sex can be distinguished based on gonad histology in animals with a minimum shell height of 9 mm. Expression of Cf‐foxl2 (C. farreri foxl2) was first observed in cytoplasm of ovarian germ and follicle cells, just after the initiation of ovarian differentiation. Results indicate that Cf‐foxl2 can be used as an early sex‐marker gene, and may have participated in the regulation of ovary differentiation in C. farreri.     

The effect of timing of extended photoperiod on growth and maturity of brook trout (Salvelinus fontinalis)

The influence of timing of extended photoperiods on growth and maturity of brook trout was investigated in a 112‐day experiment. The fish with mean initial weight of ~192 g were reared under four light regimes: one control group with natural ambient photoperiod and three groups exposed to an 18L:6D regime initiated at days 1, 23 or 46 of the growth trial. Light‐emitting diodes, with intensity of 250–1000 lux, depending on the distance from the light source, were used for extending light periods. There was a positive effect of prolonged day length on fish growth (p < 0.05), and a delay in gonad development and sexual maturity. Significantly higher numbers of sexually mature fish were found among controls groups, regardless of sex. Survival rate was not affected by light regime. This study demonstrated that the short‐term expansion of the photo period delayed maturation and increased the growth rate of brook trout.     

Effect of temperature on the efficiency of feminization of medaka (Oryzias latipes) by hormonal (estradiol) manipulation

During the critical time of sex determination, fish gonads are liable to chemicals and environmental factors that can disrupt the normal course of sex differentiation. This study was conducted to examine the possibility of a combined effect of water temperature and an exogenous steroid hormone (estradiol-17β, E₂) on the process of sex differentiation of Japanese medaka (Oryzias latipes) of the d-rR strain. Sex reversal experiments conducted at different temperatures (14, 18, 22, 26, 30, and 34 °C) and E₂ concentrations (0, 1 and 10 ppb) showed that the percentage of feminization of genotypic males into phenotypic females decreased with increasing temperature for both E₂ concentrations, suggesting a marked synergism between the two factors.     

石鲽仔、幼鱼性腺发育的组织学观察

利用组织学方法对人工培育的石鲽(Kareius bicoloratusBasiewsky)仔、幼鱼性腺发育进行研究。结果表明,性腺的发育与体长密切相关。刚孵化石鲽的原生殖细胞数目为2个,孵化3 d数目增至8个,之后经过迁移,至孵化9 d到达生殖嵴。在全长为7.2～8.5 mm(孵化9～11 d)的仔鱼中,性腺原基中的体细胞迅速增殖并包围原生殖细胞,后者在全长10～15 mm(孵化后10～35 d)的仔幼鱼中增殖成为生殖干细胞。原始性腺在全长15～30 mm(孵化40～60 d)的幼鱼中逐渐发育完善,呈细线状,位于腹腔后部中肾管下方紧贴体壁。雌性性腺最早在全长32.5 mm(孵化66 d)的个体中出现分化特征,至全长89～102 mm时雌性性腺特征完全分化。雄性性腺的分化较雌性性腺晚,最早在全长为91 mm的幼鱼中开始,至全长为114～118 mm时雄性性腺分化特征已经十分明显。     

Reproductive performance in female strains of Nile tilapia, Oreochromis niloticus

The Nile tilapia is among the most cultivated tropical fish in the world, yet little is known about the variations in reproductive characteristics among different strains. There is an interest in evaluating the reproductive performance of females of three tilapia strains. All strains were represented in equal proportion (n = 225), being named as Supreme (SUP), Premium Aquabel (PA), and Chitralada (CHI). These fish were cultivated for 7 weeks in the Aquabel Pisciculture Station (Brazil), from January to March 2011. The PA was the largest strain, with the highest values in all the morphometric parameters we analyzed (weight, total length, standard length and body depth). Among the evaluated reproductive parameters, the PA strain differed significantly from the other two in number and weight of eggs/female, volume of eggs/spawn and number of eggs/kg of fish. The percentage of non-spawning females in the SUP and CHI strains differed significantly from one another, while results for PA showed intermediate values and no significance compared to the other strains (P > 0.05). Among the reproductive parameters analyzed, the percentage of spawned females/week, more than one spawns during 7 weeks, and hatching rate showed no significant differences between strains (P > 0.05). There was a significant correlation (r = 0.69) between water temperature and percentage of spawned females/week. When evaluating the interval between two consecutive spawns, we observed that periods of 21 days were the most frequent in all strains. In conclusion, the strain that showed the best results in all morphometric and in most reproductive parameters we evaluated was the PA.     

The Influence of Sexual Maturity,Sex and Size on Quality Aspects of Frozen Argentine Hake (Merluccius hubbsi)

The Argentine hake (Merluccius hubbsi) shows a seasonal variation in its biological condition. Results of seasonal differences in gonad maturation and spawning as well as the effect on commercialization have been reported. Considering that some technological aspects and the quality of the frozen final product are dependant on the quality of raw material, the purpose of the work was to determine the influence of sexual maturity, and other biological characteristics (sex and size) of the fish on the quality of frozen Argentine hake. Quality of fish was evaluated by determination of: Total volatile bases content (TVB), Dimethylamine nitrogen content (DMA), Formaldehyde content (FA), Salt soluble protein (SSP), Drip and Sensory evaluation. Comparisons of the experimental results between the different lots studied show that contents of FA, DMA, SSP and Drip were affected by sex, size and sexual maturity. TVB content was not affected by any of the aforementioned biological factors.     

Role of sex steroids in gonadal differentiation of the mussel Choromytilus chorus (Bivalvia Mytilidae) (Molina 1782)

In Chile, the species Choromytilus chorus stands out for its ecological and economic importance. This mussel has a very particular condition in the colour of the gonad, being cream‐yellow colour the male gonad and dark brown the female which is hardly desired by the consumers. In this context, the aim of this research is to determine the role that sex steroids have in gonadal differentiation of the mussel C. chorus. For this, juveniles with sizes 15–22 mm were selected, which were subjected to acute exposure in the form of bath for 60 days to four treatments: T1: dihydrotestosterone (DHT); T2: 17β‐estradiol (E2), T3: fadrozole (F) and T4: DHT‐F (DF), plus a control without chemicals. Each treatment as well as control included three replicates with 90 individuals each. The mussels undergoing treatment with E2 had a sex ratio (male:female) of 0.47 compared to 1:1 in the control group. In contrast, in the groups treated with DHT, F and DF the sex ratio changed to 2.0, 1.60 and 1.70 respectively. In the fertilization trials, all the mussels produced functional gametes, as they were able to generate gametes that were developed to morula, and veliger larva. Nevertheless, it should be noted that the size of oocytes from the group exposed to estradiol (61.12 ± 2.40 μm) was significantly higher than in other groups. These results support the hypothesis that sex steroids would be involved in sexual differentiation of marine bivalves.     

Carotenoid pigmentation in relation to feed intake, growth and social interactions in Arctic charr, Salvelinus alpinus (L.), from two anadromous strains

Feed intake, growth and carotenoid pigmentation in 1 + Arctic charr, Salvelinus alpinus (L.), of the Hammerfest and Svalbard strains were studied in fish reared in either separate or mixed groups for 11 weeks.
Hammerfest charr grew faster than charr from the Svalbard strain at the group level. The slower overall growth in the Svalbard strain was accompanied by greater variability in feed intake and growth rates than observed amongst the fish of the Hammerfest strain. The higher incidence of bite marks amongst the Svalbard chart suggested that aggressive encounters were more frequent amongst fish of this strain. No significant differences were observed in muscle carotenoid concentration. There was, however, a highly significant positive correlation between muscle carotenoid concentration and weight gain for individual fish of both strains, irrespective of whether the fish were reared separately or in mixed groups. Estimates of muscle carotenoid retention varied from 78 to 96 mg g⁻¹, but there were no differences between strains or between fish reared in single or mixed-strain groups.
There were no differences in muscle carotenoid composition between strains or between fish reared in separate or mixed groups. Astaxanthin and idoxanthin, a metabolite of astaxanthin, were present in approximately equal proportions, and made up about 99% of the muscle carotenoid content.
The results of this study suggest that flesh pigmentation and its variability may be profoundly influenced by the level of social interactions, mediated through effects on feed intake and growth. These effects may mask genetic variations in the capacity to deposit carotenoids.     

菊黄东方鲀性腺分化的组织学观察

运用组织切片法,对菊黄东方鲀性腺分化过程进行了观察。在40日龄前,原始性腺未分化。40日龄(21.60～24.71mm),约半数个体开始形成卵巢腔,分化为卵巢;其他约半数个体的性腺仍未分化。雌鱼在60日龄(35.51～43.60mm),形成封闭的卵巢腔;70日龄(46.30～54.75mm),卵原细胞形成卵母细胞;120日龄(94.83～107.29mm),产卵板充分发育,卵巢结构已近完善。雄鱼在60日龄(35.51～43.60mm),形成精小囊及精原细胞,开始分化;90日龄(71.59～80.54mm),形成精小管和精小叶;120日龄(94.83～107.29mm),初级精母细胞形成,性腺分化基本完成。雌鱼性腺分化时期早于雄鱼。     

芳香化酶抑制剂对暗纹东方鲀CYP19A、DMRT1基因表达及性腺分化的影响

采用组织学和分子生物学方法,研究了投喂芳香化酶抑制剂来曲唑(LE)后暗纹东方鲀(Takifugu obscures)初孵仔鱼CYP19A、DMRT1基因表达以及性腺的组织学变化,以期进一步了解P450芳香化酶(P450arom)在鱼类早期性别分化过程中的作用。RT-PCR结果显示,对照组样品CYP19A和DMRT1表达显示性二态,雌性表达CYP19A基因,雄性表达DMRT1基因。LE处理组在性别分化期间,雄性样品单一表达DMRT1,雌性样品则同时表达CYP19A和DMRT1。qRT-PCR结果显示:LE处理组雌性仔鱼CYP19A基因表达被显著抑;虽然在仔鱼出膜后22d(dph)的表达水平高于9 dph,但仅为同日对照组的2.11%。LE处理组雌性样品22 dph时DMRT1基因表达量上调,至150 dph时达对照组雄性水平。55 dph的性腺组织学结果表明,LE处理可导致暗纹东方鲀稚鱼原始卵巢退化,并向功能性精巢发育。150 dph的LE处理组性腺均为精巢,并与对照组精巢发育同步。结论认为,暗纹东方鲀性腺分化期间P450arom是卵巢形成和维持发育所必须的,抑制P450arom活性可导致雌性暗纹东方鲀发生雄性化逆转。     

芳香化酶抑制剂对暗纹东方鲀CYP19A、DMRT1基因表达及性腺分化的影响

摘要: 采用组织学和分子生物学方法, 研究了投喂芳香化酶抑制剂来曲唑(LE)后暗纹东方鲀(Takifugu obscures)初孵仔鱼CYP19A、DMRT1基因表达以及性腺的组织学变化, 以期进一步了解P450芳香化酶(P450arom)在鱼类早期性别分化过程中的作用。RT-PCR结果显示, 对照组样品CYP19A和DMRT1表达显示性二态, 雌性表达CYP19A基因, 雄性表达DMRT1基因。LE处理组在性别分化期间, 雄性样品单一表达DMRT1, 雌性样品则同时表达CYP19A和DMRT1。qRT-PCR结果显示: LE处理组雌性仔鱼CYP19A基因表达被显著抑; 虽然在仔鱼出膜后22 d(dph)的表达水平高于9 dph, 但仅为同日对照组的2.11%。LE处理组雌性样品22 dph时DMRT1基因表达量上调, 至150 dph时达对照组雄性水平。55 dph的性腺组织学结果表明, LE处理可导致暗纹东方鲀稚鱼原始卵巢退化, 并向功能性精巢发育。150 dph的LE处理组性腺均为精巢, 并与对照组精巢发育同步。结论认为, 暗纹东方鲀性腺分化期间P450arom是卵巢形成和维持发育所必须的, 抑制P450arom活性可导致雌性暗纹东方鲀发生雄性化逆转。

     

Sex change in coral reef fish

Gonadal differentiation can take many forms in fish, ranging from gonochorism, where individuals directly develop as male or female and finally possess only testis or ovaries at sexual maturation, to hermaphroditism where the same individuals can produce mature male and female gametes at some time in their lives. Hermaphrodite fish are, thus, an excellent model for studying the plasticity of sex determination and differentiation in vertebrates. We have shown that sex steroids play a principal role in sex differentiation and sex change in fish. Our laboratory implements several fish models that undergo sex change from female to male or male to female or in both directions. In this review, we will briefly discuss recent advances in our understanding of the mechanism of sex change in coral reef fish.     

Growth performance of mixed sex, hormonally sex reversed and progeny of YY male tilapia of the GIFT strain, Oreochromis niloticus

The growth performance of three experimental groups consisting of mixed sex fish (control), hormone-treated fish and progeny of YY male tilapia, all originated from the genetically improved farmed tilapia (GIFT) strain was evaluated. Masculinization of sexually undifferentiated fry was achieved by providing a supplement of 5 mg of 17-α-methyltestosterone per kg of feed over a period of 21 days (after sac absorption). Both mixed sex and progeny of YY male groups were fed a standard commercial ration. Mixed sex fish did not deviate significantly ( P >0.05) from the 1:1 male to female ratio. Percentages of male averaged 75% in hormone-treated fish and 95% in YY male group over the sampling periods and at final harvest. The effect of sex on weight and length was statistically significant ( P <0.001). The model used to analyse weight and length included experimental group and sex in each culture period as the fixed effects, and replicate cages as the random effect. Over the culture period of 141 days, there were no statistical differences ( P >0.05) in body weight and length between mixed sex, hormonally treated and progeny of YY males. There were also no significant differences in level of variability in harvest weight between three groups of fish when the data were classified into five categories (=<100, 100 to <150, 150 to <200, 200 to <250 and ≥250 g). It is concluded that monosex culture of all male tilapia would be of no advantage over mixed sex culture for the GIFT strain under conditions of cages suspended in earthen ponds.