Puberty visualized: sexual maturation in the transparent Casper zebrafish.

Transparent Casper zebrafish allow studies of vertebrate sexual maturation and gonad development in vivo. Casper gonad dynamics can be observed longitudinally over time and non-invasively. Gonad maturation and reproduction are complex processes subject to disruption by endocrine-disrupting chemicals (EDCs), such as diethylstilbestrol (DES). DES was used as a 'proof of principle' to ascertain the usefulness of the Casper model to determine EDC effects on gonad maturation. Puberty onset in control juvenile Casper zebrafish (N = 43) averaged 13.2 weeks post fertilization (WPF) for females and included increased vent size, while in males puberty occurred at 11.7 WPF along with maintenance of small vents. DES treatment for 6 days in early juveniles (N = 20) induced an average delay in puberty of 5 weeks in females and 10 weeks in males. DES induced loss of breeding tubercles and vent enlargement in post-pubescent males. Puberty in control fish was correlated with an average body length of 1.7 cm for males and 1.8 cm for females. Increased testes opacity, small vent and breeding tubercles denoted male puberty. Puberty in females was defined as ovarian follicle diameters reaching 400 µm with increasingly opaque follicles and by an increased vent size. These results are like those for wild-type zebrafish and indicate that the Casper model is a useful system for studying gonad dynamics in vivo. Future use of transgenic reporter lines in Casper will allow new avenues of investigation into the reproductive biology of this vertebrate model.

Changes to sleep-wake behaviors are associated with trajectories of pubertal timing and tempo of secondary sex characteristics.

We examined relations between sleep-wake behaviors and pubertal development from age 8.5 through 15.5 years in a US-based sample of 488 boys (75% White) and 478 girls (78% White). Applying conditional nonlinear growth models to 7-waves of longitudinal data, we examined how sleep-wake behaviors are related to individual differences in the developmental timing and tempo of secondary sex characteristics. For girls, results supported the hypothesis that circadian changes in bedtimes, wake times, sleep duration, and eveningness preference were uniquely related to development of discrete aspects of secondary sex characteristics. For boys, the hypothesis was generally not supported. Different endocrine systems related to discrete secondary sex characteristics may be responsible for more relations between sleep and pubic hair development than for breast or genital development. Further research into associations between adrenarche and sleep during puberty may help researchers understand more about the origins and timing of adolescent sleep changes.

Biological sex identification in the endangered dusky gopher frog (Lithobates sevosa): a comparison of body size measurements, secondary sex characteristics, ultrasound imaging, and urinary hormone analysis methods.

BACKGROUND: Accurate sex identification techniques are important for wildlife demographic studies and for genetic management of captive breeding colonies. Various non-invasive methods for identification of biological sex in the weakly dimorphic endangered dusky gopher frog (DGF; Lithobates sevosa) were explored to support planned recovery efforts for this species including breeding and augmentation of wild populations. METHODS: Body size (snout-vent length and body weight) measurements, observation of nuptial pads, ultrasound imaging, and urinary hormone analysis for testosterone and estrone were performed on 27 male and 19 female DGFs. For each method, the mean and range of measurement values were determined for male and female DGFs housed in a captive breeding population. The ability of these methods to accurately predict the true biological sex of the individuals was assessed retrospectively. RESULTS: Body size measurements were of limited use for sex identification purposes, as males and females demonstrated overlapping body lengths and weights. Observation of the presence/absence of nuptial pads in males and females, respectively, proved to be accurate and easy to perform in most cases. Ultrasound imaging was useful for predicting the sex of female frogs, particularly when females were gravid. Commercial enzyme immunoassay kits were validated to measure urinary hormones in the DGF. Mean urinary testosterone (males: 2.22 ± 0.38 ng/ml; females: 0.92 ± 0.11 ng/ml) and estrone (males: 0.08 ± 0.01 ng/ml; females: 1.50 ± 0.39 ng/ml) concentrations were significantly (p < 0.05) different between the sexes. However, there was some overlap in hormone concentrations between the sexes. When a ratio of testosterone (T) to estrone (E) concentrations was calculated for each individual, males demonstrated significantly greater T/E ratios compared to females (p < 0.05). Use of this ratio showed greater accuracy in predicting the sex of the animal compared to using testosterone or estrone concentrations alone. CONCLUSIONS: Monitoring for presence/absence of nuptial pads and using urinary testosterone to estrone hormone ratios were the most accurate methods for identifying the biological sex of adult DGFs. Urinary hormone measurements for sex identification may be useful in other weakly dimorphic and monomorphic amphibian species in both ex situ and in situ settings.

Ontogenetic stage-specific quantitative trait loci contribute to divergence in developmental trajectories of sexually dimorphic fins between medaka populations.

Sexual dimorphism can evolve when males and females differ in phenotypic optima. Genetic constraints can, however, limit the evolution of sexual dimorphism. One possible constraint is derived from alleles expressed in both sexes. Because males and females share most of their genome, shared alleles with different fitness effects between sexes are faced with intralocus sexual conflict. Another potential constraint is derived from genetic correlations between developmental stages. Sexually dimorphic traits are often favoured at adult stages, but selected against as juvenile, so developmental decoupling of traits between ontogenetic stages may be necessary for the evolution of sexual dimorphism in adults. Resolving intralocus conflicts between sexes and ages is therefore a key to the evolution of age-specific expression of sexual dimorphism. We investigated the genetic architecture of divergence in the ontogeny of sexual dimorphism between two populations of the Japanese medaka (Oryzias latipes) that differ in the magnitude of dimorphism in anal and dorsal fin length. Quantitative trait loci (QTL) mapping revealed that few QTL had consistent effects throughout ontogenetic stages and the majority of QTL change the sizes and directions of effects on fin growth rates during ontogeny. We also found that most QTL were sex-specific, suggesting that intralocus sexual conflict is almost resolved. Our results indicate that sex- and age-specific QTL enable the populations to achieve optimal developmental trajectories of sexually dimorphic traits in response to complex natural and sexual selection.

Combined effects of binary mixtures of 17ß-estradiol and testosterone in western mosquitofish (Gambusia affinis) after full life-cycle exposure.

Estrogens and androgens are typical steroid hormones and often occur together in contaminated aquatic environments, but their mixed effects in aquatic organisms have been less well reported. In this study, the endocrine disrupting effects of binary mixtures of 17ß-estradiol (E2) and testosterone (T) in western mosquitofish (Gambusia affinis) were assessed by analyzing the sex ratio, secondary sex characteristics, gonadal histology, and transcriptional expression of target genes related to the hypothalamic-pituitary-gonadal (HPG) axis in G. affinis (from embryos) continuously exposed to E2 (50 ng/L), T (T1: 50 ng/L; T2: 200 ng/L), and mixtures of both (E2 + T1: 50 + 50 ng/L; E2 + T2: 50 + 200 ng/L) for 119 d. The results showed that exposure to E2 + T1 and E2 + T2 reduced the length ratio of ray 4/6 ratio in male G. affinis, suggesting feminized phenomenon in male G. affinis. Furthermore, 16.7-38.5 % of female G. affinis showed masculinized anal fins and hemal spines when exposed to T alone and in combination with E2. Importantly, the transcriptional levels of certain target genes related to the HPG axis were significantly altered in G. affinis following exposure to E2 and T alone and in combinations. Moreover, exposure to E2 and T in combinations can lead to combined effects (such as synergistic and antagonistic effects) on the transcriptional levels of some genes. These results collectively suggest that exposure to environmentally relevant concentrations of E2 and T alone and in mixtures can impact the endocrine system of G. affinis, and may pose potential risks in aquatic systems.

Phenotypic Mimicry Distinguishes Cues of Mating Competition From Paternal Investment in Men's Conspicuous Consumption.

Evolutionary psychologists propose that men's conspicuous consumption facilitates mate attraction because it predicts resource investment in offspring. This article elaborates on the ultimate functions of men's luxury displays based on Life History Theory. Three studies provide evidence for phenotypic mimicry, in which consumer product features mimicking male secondary sex characteristics indicate investment in mating competition, at the expense of paternal investment. Men owning shirts with larger luxury brand logos were rated higher on mating effort, lower on parental investment, higher on interest in brief sexual affairs, lower on interest in long-term committed romantic relationships, higher in attractiveness to women for brief sexual affairs, lower in attractiveness to women for long-term committed relationships, and higher in developmental environment unpredictability compared with men owning shirts displaying a smaller logo. Participants recognized the strategic use of luxury display properties across social contexts but did not consistently associate product properties with owners' physiological characteristics.

Turner syndrome with rapidly progressive puberty: a case report and literature review.

This report describes a clinically rare and atypical case of 46,X,idic(X)(q21.32)/45,X-type Turner syndrome with rapidly progressive puberty development. After 11 months of treatment with recombinant human growth hormone (rhGH), the child's height increased. After 18 months of treatment with rhGH, the child showed secondary sex characteristics. The child was followed up for 1 year after the appearance of the secondary sex characteristics, and regular menses were still present. This case indicates that modern molecular biology techniques should be used rationally to further investigate the existence of X-chromosome translocations and occult chimeras to prevent misdiagnosis.

Endocrine disrupting effects in western mosquitofish Gambusia affinis in two rivers impacted by untreated rural domestic wastewaters.

Domestic wastewaters are an important source of endocrine disrupting chemicals in the receiving aquatic environment. Most rural domestic wastewaters (RDWs) in China have been directly discharged into the aquatic environment without any treatment. Here we studied the effects of RDWs on the western mosquitofish (Gambusia affinis) from two rural rivers receiving untreated RDWs. Mosquitofish samples were collected at 5 sampling sites along two rivers during dry and wet seasons. Sex ratios, secondary sex characteristics and transcriptional levels of target genes related to the endocrine system in adult females and males were determined. In parallel, various pollutants including steroid hormones, phenolic compounds, pesticides, polycyclic aromatic hydrocarbons (PAHs) and heavy metals were measured in the water samples at all sites. The results showed that the androgenic effects in the fish were evidenced by significant increase in male to female ratio in fish populations at two sampling sites and by the presence of modified hemal spines in females at four sampling sites when compared to the reference site. The males from the two rivers had increased Vtg mRNA expressions with a maximal 6.2-fold increase relative to the reference site and a delayed development of hemal spines. The redundancy analysis (RDA) showed that some physiological parameters were related to steroid hormones, phenolic compounds and PAHs. The findings from this study suggest that RDWs can lead to masculinization in females and feminization in males.

Interpretation of fish biomarker data for identification, classification, risk assessment and testing of endocrine disrupting chemicals.

Chemical induced changes in fish biomarkers vitellogenin (VTG), secondary sex characteristics (SSC), and sex ratio indicate modes/mechanisms of action (MOAs) of EAS (estrogen, androgen and steroidogenesis) pathways. These biomarkers could be used for defining MOAs and the causal link between MOAs and adverse effects in fish for the identification of endocrine disrupting chemicals (EDCs). This paper compiled data sets of 150 chemicals for VTG, 57 chemicals for SSC and 38 chemicals for sex ratio in fathead minnow, medaka and zebrafish. It showed 1) changes in fish biomarkers can indicate the MOAs as anticipated; 2) in addition to EAS pathways, chemicals with non-EAS pathways induced changes in fish biomarkers; 3) responses of fish biomarkers did not always follow the anticipated patterns of EAS pathways. These responses may result from the interaction of chemical-induced multiple MOAs and confounding factors like fish diet, infection, culture conditions, general toxicity and stress response. The complex response of fish biomarkers to a chemical of interest requires EDC testing at multiple biological levels. Interpretation of fish biomarker data should be combined with relevant information at different biological levels, which is critical for defining chemical specific MOAs. The utility of fish biomarker data for identification, classification, PBT assessment, risk assessment, and testing of EDCs in the regulatory context was discussed. This paper emphasizes the importance of fish biomarker data in the regulatory context, a weight of evidence approach for the interpretation of fish biomarker data and the need for defining levels of evidence for the identification of EDCs.

Fish biomarkers for regulatory identification of endocrine disrupting chemicals.

Demonstrating chemical-induced adverse effects, endocrine mechanisms/modes of action (MOAs) and their causal link is needed for regulatory identification of endocrine disrupting chemicals (EDCs). This paper addresses critical issues over MOAs and their causal link to changes in endpoints. Vitellogenin (VTG), secondary sex characteristics (SSC), and sex ratio (also an apical endpoint) are indicative of chemicals interfering with EAS (estrogen, androgen and steroidogenesis) pathways. These biomarkers, however, can be changed by non-EAS chemicals, systemic toxicity and the stress response. Examples are shown that proving causal link between MOAs and changes in endpoints may be difficult for regulatory identification of EDCs. The paper concludes that both in vitro and in vivo data are needed to define MOAs for regulatory identification of EDCs. Further development of guidance documents for data interpretation and for defining the level of evidence is needed for regulatory identification of EDCs.