Genetic, abiotic and social influences on sex differentiation in cichlid fishes and the evolution of sequential hermaphroditism

Genetic and environmental factors may interact to control sex determination in fishes. A common pattern of initial female differentiation and subsequent male transformation before maturation in non‐hermaphroditic fishes and after maturation in sequentially hermaphroditic fishes has suggested that changes in developmental timing may be responsible for the evolution of various expressions of sexual lability. Sequential hermaphroditism is rare in freshwater fishes, but investigators report degrees of sexual lability at four distinct life stages in cichlid fishes. Some cichlids undergo genetic sex determination and are not labile. Lability at the larval stage allows temperature or pH to determine sex. Social interactions apparently determine sex at the juvenile stage in the Midas cichlid (Amphilophus citrinellus). Most reports of post‐maturational sex change in cichlids are anecdotal or unsubstantiated. The common occurrence of same‐sex spawning suggests that many species are incapable of sex change. Sequential hermaphroditism is concluded not to be typical, except for the checkerboard cichlid (Crenicara punctulata), which regularly undergoes functional female‐to‐male transformation. Expression of sexual lability at four life stages in one family of fishes corroborates a role for developmental timing in the evolution of sequential hermaphroditism as well as environmentally controlled sex determination. The broad phylogenetic distribution of sexual lability in cichlids indicates that processes capable of producing sex change are generally present. The rarity of sequential hermaphroditism in cichlids and possibly other freshwater fishes is likely due to unpredictability of food and therefore potential mate distributions compared with coral reef habitats.     

Gonadotropin-releasing hormone and gonadotropin in goldfish and masu salmon

Reproductive activities in vertebrates are regulated by an endocrine system, consisting of the brain-pituitary-gonad axis. In teleosts, gonadotropin-releasing hormone (GnRH) in the brain stimulates gonadotropin (GTH) release in the pituitary gland, but because of lack of the portal vessel, it is not known when and how much GnRH is released for the regulation of GTH release. There are multiple molecular types of GnRH in teleosts and several distinct populations of GnRH neurons in the brain. However, we do not know which types and populations of GnRH neurons regulate reproductive activities. Here we summarize our recent studies on GnRH and GTH in masu salmon Oncorhynchus masou and goldfish Carassius auratus. Immunocytochemistry showed the location and molecular types of GnRH neurons. Salmon (sGnRH) and chicken-II GnRH (cGnRH-II) neuronal fibers were widely distributed in the brain of both masu salmon and goldfish. Only sGnRH fibers were observed in the pituitary of masu salmon, whereas both sGnRH and cGnRH-II fibers were observed in the goldfish pituitary, indicating that species specific GnRH profiles are involved in the regulation of pituitary function in teleosts. A series of experiments in masu salmon and goldfish suggest that among GnRH neuron populations GnRH neurons in the ventral telencephalon and the hypothalamus regulate GTH release, and that GnRH of the terminal nerve origin is not essential to gonadal maturation and ovulation. The biological function of other GnRH neurons remains unkown. Two GTHs appear to be characteristic of teleost; however, regulation of reproduction by these GTHs is a question that remains to be elucidated. In salmonid species, it is proposed that GTH I stimulates early gonadal development, whereas GTH II acts in later stages. When GTH expression was examined in goldfish, both GTH I and II mRNA levels in the pituitary gland showed increases in accordance with gonadal development, unlike the sequential expression of GTH subunits in salmonids. The expression of these GTH subunit mRNAs were affected by water temperature, starvation, and steroid hormones in goldfish, but in what manner these two GTHs regulate gonadal development remains to be clarified.     

雌雄同体鱼类性别分化及性转变研究进展

在脊椎动物中,鱼类具有多样的性别分化方式,大致可分为雌雄异体、雌雄同体以及单性生殖3类.一般情况下,鱼类性别决定后,性腺可分化为卵巢或精巢,并且在整个生命周期内保持不变.而在雌雄同体鱼类,其性别可以从雌性转变为雄性、雄性转变为雌性或者在雌雄两种性别间进行多次转变.雌雄同体鱼类具有多种性别转变形式,是研究脊椎动物性别决定...     

Seasonal reproductive cycle of Waigieu seaperch (Psammoperca waigiensis)

Predictive and reliable parameters of reproductive status are integral aspects of sustainable fisheries and aquaculture management. These parameters are also important for an accurate evaluation of the effects of different treatments on sexual maturation in fish farming. In the present study, we have characterized the seasonal reproduction profile and described changes in sex steroids in relation to gonadal maturation and development in female and male Waigieu seaperch (Psammoperca waigiensis). The experimental period covered a full calendar year (January–December). In males and females, we observed that plasma sex steroid hormones [oestradiol‐17β (E2), testosterone (T), 11‐ketotestosterone (11‐KT) and progesterone (P)] levels showed monthly fluctuations during the spawning period. Particularly, plasma steroid hormone levels were positively associated with gonadosomatic index values. In addition, high levels of plasma steroid coincided with recruitment of oocytes into yolk accumulation in females. The main spawning period occurred between April and October in females, and between March and November in males. The non‐aromatizable androgen, 11‐KT is generally believed to be the active male‐specific androgen in teleosts, and is associated with the process of spermiation, development of secondary sexual characteristics and regulation of male reproductive behaviour in most teleost species. In this study, we found relatively high amounts of 11‐KT in females between May and December, suggesting an integral role in the maturation process, also for the females. A rapid peak in plasma P level was observed in November and suggests significant roles during post‐spawning and/or resting periods in both female and male fish. Furthermore, all oocyte developmental stages were present within the same sampling month and also within the spawning period, demonstrating the gamete group asynchronous developmental strategy. Overall, Waigieu seaperch showed strong seasonality in reproductive development with corresponding sex steroid patterns. The data presented in this study may contribute to the understanding of the reproductive endocrinology of a tropical marine finfish with increasing industrial prospects and sustainable aquaculture of this species in a developing country, such as Vietnam.     

Leptin in fish: possible role in sexual maturation in male Atlantic salmon

Leptin plays an important role in energy homeostasis and reproductive function in mammals, while its function in fish is still poorly understood, especially its role in reproduction. In the present study, leptin gene expression and circulating leptin plasma levels were measured during sexual maturation in male Atlantic salmon parr. In maturing male salmon, higher hepatic leptin (lepa1) gene expression levels were observed during mid-spermatogenesis compared to immature fish. An upregulation of leptin during sexual maturation has also been observed in a few other teleost species. The physiological significance of elevated leptin levels during the maturational process in teleosts remains to be explored.     

Reproductive biology of freshwater fishes from the Venezuelan floodplains

This review describes the endocrine changes that occur during the annual reproductive cycle of Pygocentrus cariba, Pimelodus blochii, and Oxydoras sifontesi and their relationships with the environmental characteristics of Venezuelan floodplains. Most reproductive studies of teleosts have focused on changes that occur during annual cycles in temperate species but, in tropical fish, this has been examined less frequently. P. cariba, P. blochii, and O. sifontesi are seasonal breeders widely distributed along the Orinoco River. Under natural conditions they have an annual gonadal cycle closely related to changes in the annual hydrology cycle of the Orinoco River which defines two seasons on the floodplain: inundation and isolation. The reproductive cycle of these species seems to be controlled by cues from the external environment. Relevant data about gonadal maturation, for example gonadosomatic index and sexual hormones secretion, are contrasted. The role of catecholamines in neuroendocrine control of the reproductive axis is also considered in this work.     

Regulation of forebrain and midbrain GnRH neurons in juvenile teleosts

Three molecular species of gonadotropin releasing hormone (GnRH) mRNA-containing neuronal populations (terminal nerve: salmon-GnRH; preoptic area: seabream-GnRH; midbrain: chic- ken-GnRH-II) have been localized in teleosts. While the termi- nal nerve GnRH neurons originate from the olfactory placode, a separate intracerebral source of origin for preoptic and midbrain neurons is possible. The preoptic GnRH neurons are regulated by gonadal steroids and gonadal maturation, however, the regulation and role of terminal nerve and midbrain GnRH neurons in teleosts reproduction is speculative and debatable.     

The remarkable reproductive diversity of teleost fishes

Teleosts display the most striking reproductive diversity of all vertebrates. However, no convincing hypothesis has yet been proposed to explain why they have evolved this remarkable variability in their modes of reproduction. Some of the features of the reproductive biology of teleosts are briefly reviewed, and unique characteristics of the group that may have made possible the evolution of their remarkable reproductive diversity are considered. These include whole‐genome duplication, the mode of differentiation of the gonads and the organization of the brain–pituitary relationship.     

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.     

Morphological skin colour changes in teleosts

Morphological skin colour change in fish is often referred to in the sole context of background adaptation. It is becoming increasingly apparent that it is a broad phenomenon elicited by a variety of factors. To date, no review has attempted to integrate the different types of morphological colour changes occurring in teleosts, their ecological origins and the regulatory mechanisms involved, often restricting the view on the subject. First, the origin of skin colour is addressed in teleosts including chromatophore type and distribution, pigment biosynthetic pathways and their interactions to one‐another. Second, the different types of morphological colour changes occurring in teleosts are categorized and a key distinction is made between proximate and ultimate morphological colour changes. These are defined respectively as the change of phenotype during an established life‐stage in response to environmental interactions and during the transition between two developmental‐stages phenotypically pre‐adapted to their ancestral ecosystems. Nutrition and UV‐light are primary factors of proximate morphological colour changes beyond the control of the organism. By contrast, background light conditions and social interactions are secondary proximate factors acting through the control of the organism. Highly diversified among teleosts, ultimate morphological skin colour changes are presented in term of alterations in skin structure and pigment deposition during metamorphosis in different species. Finally, the physiological and endocrine mechanisms regulating both proximate and ultimate morphological colour changes are reviewed.     

Aromatase (P450arom) and 11β-hydroxylase (P45011β) genes are differentially expressed during the sex change process of the protogynous rice field eel, monopterus albus

Steroids are known to play a crucial role in gonadal sex differentiation in many non-mammalian vertebrates, but also in the gonadal sex change of hermaphroditic teleosts. We investigated the expression of two genes encoding key steroidogenic enzymes, i.e., cytochrome P450 aromatase (P450arom) and cytochrome P45011β-hydroxylase (P45011β), during the sex change of the protogynous rice field eel, Monopterus albus. Using RT-PCR with degenerate primers, we cloned rice field eel homologous fragments for both genes (rcP450arom and rcP45011β) as indicated by the high level of homology with P450arom and P45011β sequences from various vertebrates. Gonadal expression of rcP450arom and rcP45011β mRNA levels were then assessed during the sex change by semi-quantitative RT-PCR and a real-time RT-PCR. rcP450arom was predominantly expressed in ovary, much less in ovotestis, and barely in testis. Conversely, P45011β was markedly up-regulated at the onset of testicular development. These findings underline that regulation of steroidogenesis is an important process in the sex change of protogynous rice field eel, and they clearly indicate that the concomitant down-regulation of P450arom and up-regulation of P45011β are of pivotal importance to the sex change of this species.     

The influence of reproductive status on the stimulatory action of N-methyl-D,L-aspartate on growth hormone secretion, in vitro in rainbow trout, Oncorhynchus mykiss

The glutamate agonist, N-methyl-D,L-aspartate (NMA) stimulates the secretion of growth hormone (GH) from pituitary fragments in vitro and increases plasma GH levels in vivo in rainbow trout, Oncorhynchus mykiss (Flett et al. 1994; Holloway and Leatherland 1997a,b); however gonadal steroid hormones appear to modulate this response in experimental situations. This study examines whether steroid hormones also modulate the GH-regulatory actions of NMA during the normal reproductive cycle of rainbow trout by examining the relationship between the stage of sexual maturation and the pituitary release of GH in vitro in response to an NMA (10^-8 M) challenge. NMA had no effect on mean GH release from the pituitary glands of fish that were immature (GSI <1.0), from males during early development (GSI 1.0-3.0), or from sexually mature males (with free running milt) and females (ovulated). However, NMA significantly increased GH release from pituitary glands taken from females during the early stages of gonadal growth (GSI 1.0-9.0) and from males and females sampled during the later stages of gonadal growth (males GSI 3.01-6.0; females GSI 9.01-15.0). The GH-stimulatory action of NMA in males and females progressed to a maximum effect during the late stages of gonadal growth, and disappeared in ovulated females and free running males. Moreover, in female fish, the maximal GH release in response to the NMA challenge is positively correlated with plasma 17β-estradiol levels; no such correlation was evident for plasma testosterone levels in males. Changes in the GH response to NMA during maturation while gonadal steroid levels fluctuate provides further evidence to suggest that the effects of NMA on GH secretion are intimately linked to endogenous gonadal steroid hormone levels. This revised version was published online in July 2006 with corrections to the Cover Date.     

中华鳖Gper基因克隆、表达及其在精巢中的功能

G蛋白偶联雌激素受体（Gper）是一种膜雌激素受体,介导雌激素的非基因组途径。为研究G蛋白偶联雌激素受体基因（ cDNA全长序列,利用qRT-PCR分析其在不同组织及胚胎不同发育阶段的性腺中的表达模式,并通过来曲唑（letrozole）和Gper抑制剂G-15处理雄鳖初步分析Gper在精巢中的作用。结果显示,中华鳖 cDNA序列全长2023 bp,包含705 bp 5''非编码区、241 bp 3''非编码区和1077 bp开放阅读框,编码358个氨基酸,其氨基酸序列上有7个跨膜结构域和Asp-Arg-Tyr（DRY）三联体结构,基因编码蛋白分子量为41.084 kD,等电点为6.844。表达量变化呈相同趋势：16期表达量最高,随着性腺分化过程表达量显著降低。Letrozole处理组中2表达量明显升高;G-15处理组精巢中,精子发生与促细胞凋亡相关基因表达量显著升高。结果表明,可能参与中华鳖性腺分化早期过程,并调控雄性生殖细胞增殖。     

九孔鲍性腺发育过程中消化酶活力的变化

用酶学方法研究了性腺发育过程中九孔鲍肝胰腺和消化道中蛋白酶、淀粉酶和纤维素酶活力的变化。试验结果表明,雌、雄九孔鲍各消化酶活力均随着性腺发育呈先升后降的变化趋势。在性腺发育早期和晚期酶活力较低,在性腺发育中期酶活力显著升高,且均在成熟前期达到最高值。九孔鲍繁殖期各消化酶的活力分布存在器官特异性和性别差异,九孔鲍的淀粉酶和纤维素酶活力在各性腺发育期均表现为肝胰腺消化道,雌鲍雄鲍。同种消化器官内各消化酶的活力也存在差异,雌、雄九孔鲍繁殖期肝胰腺和消化道内各消化酶的最大活力均为:纤维素酶淀粉酶蛋白酶。     

Hermaphroditism in Cultured Chinese Black Sleeper (Bostrichthys sinensis L.)

Anatomical studies showed that all collected feral fish of Chinese black sleepers (Bostrichthys sinensis L.) were gonochoristic. In contrast, an average of 12.4% of cultured fish appeared to be hermaphroditic. The shape of the genital papilla of hermaphroditic specimens resembled that of gonochoristic males. The gonadosomatic index (I_G) of hermaphroditic fish varied from 0.24 to 4.48%, while that of gonochoristic conspecifics varied from 6.1 to 12.2% for females and 0.11 to 0.14% for males. Histological studies revealed that the sex organs of hermaphrodites consisted of ovarian, testicular, and seminal vesicle tissues, located in the central, peripheral, and caudal region of the gonads, respectively. During the spawning season, ovarian tissue contained poorly developed oocytes, that is, hydrated oocytes were absent and only a few degenerated vitellogenic oocytes were present. Testicular tissue, however, contained fully developed spermatozoa. The fertilization rate of these spermatozoa was significantly lower than those of gonochoristic conspecifics. It is concluded that part of the cultured stock of Chinese black sleeper became hermaphroditic during culture and that the hermaphrodites are able to function as males. The lowered fertilization rate, however, indicates that hermaphroditism in cultured fish of this species is disadvantageous to reproduction.     

Hormones, pheromones and reproductive behavior

Fish commonly use reproductive hormones (steroids and prostaglandins) both as endogenous signals between reproductive tract and brain and as exogenous signals (hormonal pheromones) that synchronize gamete maturation and/or spawning interactions between and among conspecifics. This dual function for hormonal products not only extends traditional concepts that sex hormone actions are limited to reproductive synchrony within the individual, but also implies we are unlikely to achieve a comprehensive understanding of reproductive function in any fish without knowledge of both the endogenous and exogenous actions of its hormones and related released compounds. Such knowledge is beginning to accumulate for several species (e.g. goldfish, Atlantic salmon), but even here is far from complete. Moreover, because hormonal pheromone studies have focused on oviparous gonochorists with relatively simple reproductive strategies, we know nothing about the potential hormonal pheromone functions of the numerous species with sequential hermaphroditism or alternative male strategies, or the possible changes in pheromone function associated with the numerous transitions from oviparity to viviparity. Given the insights we have gained from studies of traditional species, it seems certain that expanding hormonal pheromone research to non-traditional species exemplifying the diverse nature of fish mating systems will similarly enrich our understanding of fish reproductive function.     

The effects of salinity on reproductive development and egg and larvae survival in the spotted scat Scatophagus argus under controlled conditions

In the present study, we report the first successful instance of controlled reproduction in Scatophagus argus, which has recently emerged as a new aquaculture resource. The controlled reproduction process for S. argus was optimized with regard to salinity acclimation. Gonadal maturation was affected by salinity in both sexes. Levels of plasma 17β‐estradiol (E₂) and 11‐ketotestosterone (11‐kT) were salinity dependent and increased significantly with the duration of acclimation. Plasma levels of gonadal steroids were higher in fish held at 25‰ salinity. The highest gonadosomatic indices (GSI), 15.1 ± 1.6 in the female and 6.4 ± 1.2 in the male, were also observed at 25‰ salinity. Nevertheless, the optimal salinity for S. argus embryonic development and larval culture was 15‰. Thus, the salinity requirement for gonadal maturation and early development are quite different. The use of advanced reproductive technologies combining salinity acclimation and stimulation of luteinizing hormone‐releasing hormone analog (LHRH‐A₂) resulted in a fertilization rate of 83.2%–91.3% and embryonic survival rates of over 90%. Embryos of S. argus at the 2‐cell, blastula, gastrula and pharyngula stages were observed. Most embryos hatched after 21.0 hr of incubation at 28.0 ± 1.0°C. The development of larvae into juveniles was completed at 40–45 days posthatch (dph). In this study, we provide information about the controlled reproduction of S. argus and identify the optimal environmental parameters for S. argus embryonic and larval culture, with the aim of developing reliable reproductive techniques for its mass production.     

金钱鱼孕激素受体基因的克隆、鉴定与表达研究

核受体(Pgr),隶属于核受体超家族,由核受体亚家族3C组成员3基因编码.孕酮通过结合Pg r介导其在脊椎动物繁殖过程中的生理学功能,因此,研究金钱鱼核受体基因(pg r基因)在不同组织及不同发育时期性腺中的表达模式,有助于为金钱鱼的繁殖提供理论依据.笔者克隆金钱鱼的pg r基因,用Megalign软件比较Pgr氨基酸...     

Reproductive biology of Cyprinella camura, the bluntface shiner, in Morganfork Creek, southwestern Mississippi

Abstract– I collected bluntface shiners, Cyprinella camura , once or twice monthly from February to November 1991 in Morganfork Creek, southwestern Mississippi. Examinations of mean largest oocyte diameter (LOD), female mean relative gonadal index (RGI), ovarian developmental condition, and male secondary sexual characteristics indicated that C. camura spawned from late March through mid-August. There were significant positive correlations between mean LOD and mean daylength, mean maximum temperature, and mean minimum temperature (environmental variable means were calculated for the 28 day period prior to each sampling date). Mean day length and mean RGI were significantly positively correlated, but there was no significant relationship between mean maximum and minimum temperatures and mean RGI. Clutch size, RGI, and mean mature oocyte diameter (MMOD) of mature females were significantly correlated with length (standard length). Mean length, clutch size, and RGI of mature females in individual collections decreased significantly over the reproductive season. The corresponding decrease in mean MMOD was not significant.