Southern flounder (Paralichthys lethostigma) exhibit environmental sex determination (ESD), where environmental factors can influence phenotypic sex during early juvenile development but only in the ...presumed XX female genotype. Warm and cold temperatures masculinize fish with mid-range conditions producing at most 50% females. Due to sexually dimorphic growth, southern flounder fisheries are dependent upon larger females. Wild populations could be at risk of masculinization from ESD due to globally increasing water temperatures. We evaluated the effects of habitat and temperature on wild populations of juvenile southern flounder in North Carolina, USA. While northern habitats averaged temperatures near 23 °C and produced the greatest proportion of females, more southerly habitats exhibited warmer temperatures (>27 °C) and consistently produced male-biased sex ratios (up to 94% male). Rearing flounder in the laboratory under temperature regimes mimicking those of natural habitats recapitulated sex ratio differences observed across the wild populations, providing strong evidence that temperature is a key factor influencing sex ratios in nursery habitats. These studies provide evidence of habitat conditions interacting with ESD to affect a key demographic parameter in an economically important fishery. The temperature ranges that yield male-biased sex ratios are within the scope of predicted increases in ocean temperature under climate change.
For decades, it was widely assumed that glucocorticoids (GCs) work solely through changes in gene expression to exert their physiological actions, a process that normally takes several hours to ...occur. However, recent evidence indicates that GCs might also act at the membrane through specific receptors to exert multiple rapid effects on various tissues and cells. GCs modulate hormone secretion, neuronal excitability, behavior, cell morphology, carbohydrate metabolism and other processes within seconds or minutes. These early actions occur independent of the genome and are transduced by the same biochemical effector pathways responsible for mediating rapid responses to neurotransmitters. The biological significance of most rapid GC effects are not well understood, but many might be related to the important functions that this hormone plays in modulating stress responses.
IGF-I is a mitogenic polypeptide that is an important regulator of growth in fish. The potential of IGF-I mRNA abundance as a rapid growth indicator in the Nile tilapia,
Oreochromis niloticus, was ...evaluated. Hepatic IGF-I cDNA was isolated and partially cloned. The partial sequence having 539 bases encodes for the signal peptide, mature protein and a portion of the E domain. The deduced 68 amino acid sequence for mature IGF-I showed 84–90% and 77–79% sequence identity with fish and mammalian counterparts, respectively. The deduced amino acid sequence for domains B and A was most conserved (93–97%) relative to other fishes. A sensitive TaqMan real time qRT-PCR assay for
O. niloticus was developed based on the mature IGF-I peptide for measures of hepatic IGF-I mRNA levels. Hepatic IGF-I mRNA levels were found to be significantly correlated with growth rate of fish reared under different feeding regimes and temperature conditions. Higher feed consumption and water temperature produced faster-growing fish and increased hepatic IGF-I mRNA expression. These findings suggest that hepatic-derived IGF-I plays a key role in controlling growth in
O. niloticus and indicates that IGF-I mRNA quantification could prove useful for the rapid assessment of growth rate in this species.
Highlights • We review the regulation of prolactin cell function and modulation of osmoreception in the Mozambique tilapia. • We describe the endocrine regulators of prolactin release and mRNA ...levels. • We provide an updated discussion on the role of hormones and extracellular osmolality in the regulation of prolactin cells. • We conclude that changes in extracellular osmolality are sufficient, by themselves, to regulate prolactin cell function.
Environmental sex-determination (ESD) is the phenomenon by which environmental factors regulate sex-determination, typically occurring during a critical period of early development. Southern flounder ...(Paralichthys lethostigma) exhibit temperature-dependent sex-determination that appears to be restricted to the presumed XX female genotype with the extremes of temperature, both high and low, skewing sex ratios toward males. In order to evaluate other environmental factors that may influence sex-determination, we investigated the influence of background color and cortisol on sex-determination in southern flounder. Experiments involving three sets of tanks, each painted a different color, were conducted at different temperatures using southern flounder of mixed XX–XY genotype. The studies involved rearing juvenile southern flounder in either black, gray, or blue tanks and sex-determination was assessed by gonadal histology. In both studies, blue tanks showed significant male-biased sex ratios (95 and 75% male) compared with black and gray tanks. The stress corticosteroid cortisol may mediate sex-determining processes associated with environmental variables. Cortisol from the whole body was measured throughout the second experiment and fishes in blue tanks had higher levels of cortisol during the period of sex-determination. These data suggest that background color can be a cue for ESD, with blue acting as a stressor during the period of sex-determination, and ultimately producing male-skewed populations. In a separate study using XX populations of southern flounder, cortisol was applied at 0, 100, or 300 mg/kg of gelatin-coated feed. Fish were fed intermittently prior to, and just through, the period of sex-determination. Levels of gonadal P450 aromatase (cyp19a1) and forkhead transcription factor L2 (FoxL2) messenger RNA (mRNA) were measured by qRT–PCR as markers for differentiation into females. Müllerian-inhibiting substance mRNA was used as a marker of males’ gonadal development. Control fish showed female-biased sex ratios approaching 100%, whereas treatment with 100 mg/kg cortisol produced 28.57% females and treatment with 300 mg/kg cortisol produced only 13.33% females. These results suggest that cortisol is a critical mediator of sex-determination in southern flounder by promoting masculinization. This linkage between the endocrine stress axis and conserved sex-determination pathways may provide a mechanism for adaptive modification of sex ratio in a spatially and temporally variable environment.
Flounder of the genus
Paralichthys exhibit a unique mode of sex determination where both low and high temperatures induce male-skewed sex ratios, while intermediate temperatures produce a 1:1 sex ...ratio. Male differentiation is thus easily induced in genetic females creating a combination of genetic (GSD) and environmental sex determination (ESD). Since male flounder become reproductively fit at substantially smaller body sizes than females, temperature or other environmental variables that elicit lower growth rates may also influence sex differentiation toward male development. This review covers our current knowledge of sex determination and differentiation in flatfishes including possible adaptive significance of ESD and involvement of factors such as aromatase (
cyp19).
Southern flounder (
Paralichthys lethostigma) support valuable North American fisheries and show great promise for aquaculture. Because females grow faster and reach larger adult sizes than males, ...monosex culture of females is desirable for commercial operations. A detailed understanding of sexual development and its timing is critical to control sex and optimize culture. Structural and cellular sex-distinguishing markers were identified histologically, and then used to describe ovarian development in female and testicular development in male flounder. In presumptive ovaries of southern flounder, development of an ovarian cavity first occurs in fish ranging from 75 to 100 mm total length (TL). This is considerably delayed relative to that observed in the Japanese congener,
Paralichthys olivaceus, where an ovarian cavity is seen in fish as small as 40 mm TL. The smallest southern flounder that possessed primary oocytes in the early perinucleolus stage was 115 mm TL. In presumptive testes, the formation of seminiferous tubules first occurs in fish of approximately 100 mm TL. Spermatogonia remained quiescent until most fish were over 100 mm TL. Overall, gonads from southern flounder greater than 120 mm TL commonly possess gonial cells undergoing meiosis, clearly differentiating sex. The effect of temperature on sex determination in southern flounder was addressed in a separate experiment. Juvenile southern flounder were grown at 18, 23, or 28°C for 245 days. High and low temperatures induced phenotypic sex reversal in juvenile southern flounder, producing a higher proportion of males (96% males at high temperature,
P<0.001, 78% males at low temperature,
P<0.01). Raising southern flounder at the midrange temperature held sex ratios close to 1:1. Sex ratios from these trials suggest that southern flounder possess a temperature-sensitive mechanism of sex determination similar to that shown for
P. olivaceus, but possibly shifted towards warmer temperatures. These findings indicate that sex differentiation in southern flounder is distinguishable in most fish by 100–120 mm TL and that sex determination is sensitive to temperature. This information is critical to the development of strategies to maximize the number of faster-growing females for commercial flounder culture.
Fish represent the most diverse group of animals in the vertebrate phylum. The more than 25,000 species are characterized by an array of anatomical, biochemical, physiological, and behavioral ...repertoires. For this reason, it is difficult to develop a comprehensive guideline on the care and use of fishes. Institutional animal care and use committees (IACUCs) meet the challenge of ensuring adequate fish welfare using guidelines (Animal Welfare Act AWA and Public Health Service PHS Policy and their guides) derived mainly from the care and use of mammalian species, which may not be optimal for regulating fish research, teaching, or extension activities. Discussion focuses on various issues that often confront IACUCs in meeting regulatory requirements while assuring proper fish welfare. Issues include questions concerning animal tracking and inventory, utilization of fisheries bycatch, facility inspections in remote locations, and euthanasia. Common sense solutions appropriate for field and laboratory fish activities are suggested, which should help investigators, IACUCs, and regulatory agencies meet PHS and AWA objectives.
Effective methods for induction of diploid gynogenesis in North American flounder of the genus
Paralichthys are needed to initiate monosex culture, which will allow growers to take advantage of the ...more rapid growth and larger size attained by females. To test methods for inducing diploid gynogenesis in southern flounder (
Paralichthys lethostigma) using homologous sperm, four treatments, named for their expected outcome, were employed: haploid, diploid, triploid, and gynogenetic diploid. Diploid gynogenesis was induced by activating egg development with UV-irradiated flounder sperm (70 J/cm
2) for 3–4 min in seawater, and then subjecting the eggs to cold shock in 0–2 °C seawater for 45–50 min. Cold shock was used to prevent extrusion of the second polar body. Control treatments omitted one or more of these steps to separately assess the effectiveness of UV irradiation and cold shock. Larvae were observed for physical abnormalities and then histologically processed for ploidy determination. Haploid larvae exhibited abnormal external morphology while diploid, gynogenetic diploid, and triploid larvae showed normal morphologies. Cross-sectional areas of erythrocyte nuclei were measured for larvae in each treatment group and significant differences were found. Nuclear areas for treatment groups corresponded to predicted ploidy (triploid>diploid>haploid) and did not differ between normal diploid controls and gynogenetic diploids. These results suggest that the procedures of sperm irradiation and egg cold shock successfully generated gynogenetic diploids. Due to the low volumes of semen produced by male flounder, and to eliminate any potential genetic contribution by homologous sperm, activation of flounder eggs with heterologous sperm was also investigated. Induction of diploid gynogenesis was successful when flounder eggs were fertilized with irradiated (50 J/cm
2) sperm from striped mullet (
Mugil cephalus), and then cold shocked. This work provides procedures for induction of diploid gynogenesis in southern flounder using homologous and heterologous sperm, and validates a method for verification of ploidy in larval fish.
Abstract The regulation of growth hormone (GH) secretion by ghrelin during variable metabolic states is poorly understood. We examined plasma GH and ghrelin in hybrid striped bass (HSB) undergoing ...seasonally-based feeding and temperature manipulations. Fasting for 21 days (d) at 24 °C resulted in catabolism and up-regulation of plasma GH and ghrelin relative to fed controls. Continued fasting during cold-banking (14 °C, 90d) resulted in a further 43-fold increase in ghrelin while GH remained elevated. A subsequent 19 day refeeding period at 24 °C elicited hyperphagic and compensatory growth responses, accompanied by declines in ghrelin and GH. We then tested the role of ghrelin in stimulating GH release in vivo and in vitro . Intraperitoneal injections of ghrelin resulted in dose-dependent increases in plasma GH after 6 hours (h). Ghrelin also increased GH release from HSB pituitaries during 6 h incubations. Lastly, we assessed how metabolic state, ghrelin and insulin-like growth factor-I (IGF-I) affect in vitro pituitary GH release. Spontaneous GH release was 5.2-fold higher from pituitaries of fasted compared with fed animals. Ghrelin was equally effective in stimulating GH release from pituitaries of fed and starved animals, while it was ineffective in enhancing GH release from pituitaries of starved (21d) then refed (4d) HSB. Incubation with IGF-I inhibited GH release regardless of metabolic state. These studies are the first to show that seasonally-based periods of feed deprivation and low temperature yield sustained increases in GH secretion that are likely mediated, at least partially, through elevated ghrelin, reduced IGF-I negative feedback and fasting-induced spontaneous GH release.
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