Sex steroid dynamics during embryogenesis and sexual differentiation in Eurasian perch, Perca fluviatilis

It is widely accepted that sex steroid hormones play an important and a specific role during the process of sex differentiation in fish. In order to describe the role of the three main sex steroid hormones (testosterone--T, 17beta-estradiol--E2 and 11keto-testosterone--11KT) during embryogenesis and sex differentiation in Eurasian perch, Perca fluviatilis, eggs, larvae and juveniles originating from two mixed-sex and two all-female progenies were regularly sampled from fertilization to hatching (D0) and from hatching to day 70 post-hatching (D70). Just after spawning, a significant amount of sex steroids [T (1634.2pgg(-1)), E2 (554.4pgg(-1)) and 11KT (1513.2pgg(-1))] was measured in non-fertilised eggs suggesting a maternal transmission of these steroids. From D2 to D70 post-hatching, E2 levels were significantly higher in mixed-sex progenies (median: 725.7pgg(-1)) than in all-female progenies (156.2pgg(-1)) and significantly increased after the onset of the histological differentiation of the gonad in both progenies (D35). Levels of 11KT were significantly higher in mixed-sex (median: 431.5pgg(-1)) than in all-female progenies (below the limit of assay detection) and significantly increased at D35 in all-female progenies (median value: 343.2pgg(-1)). Mean 11KT to E2 ratio was six-fold higher in mixed-sex progenies (1.35) than in all-female progenies (0.24). The data suggest that the 11-oxygenated androgen (11KT) plays a major role in the male differentiation process, and that sex differentiation in Eurasian perch is probably determined by the 11KT to E2 ratio

DNA methyltransferases and stress-related genes expression in zebrafish larvae after exposure to heat and copper during reprogramming of DNA methylation

DNA methylation, a well-studied epigenetic mark, is important for gene regulation in adulthood and for development. Using genetic and epigenetic approaches, the present study aimed at evaluating the effects of heat stress and copper exposure during zebrafish early embryogenesis when patterns of DNA methylation are being established, a process called reprogramming. Embryos were exposed to 325 μg Cu/L from fertilization (<1 h post fertilization - hpf) to 4 hpf at either 26.5 °C or 34 °C, followed by incubation in clean water at 26.5 °C till 96 hpf. Significant increased mortality rates and delayed hatching were observed following exposure to combined high temperature and Cu. Secondly, both stressors, alone or in combination, significantly upregulated the expression of de novo DNA methyltransferase genes (dnmt3) along with no differences in global cytosine methylation level. Finally, Cu exposure significantly increased the expression of metallothionein (mt2) and heat shock protein (hsp70), the latter being also increased following exposure to high temperature. These results highlighted the sensitivity of early embryogenesis and more precisely of the reprogramming period to environmental challenges, in a realistic situation of combined stressors

Reproductive Cycle and Plasma Levels of Sex Steroids in Female Eurasian Perch Perca Fluviatilis

From April 1995 to April 1996, the annual reproductive cycle of the Eurasian perch Perca fluviatilis was studied at the Fishfarming Lindre Center (Moselle, France). At monthly intervals (at intervals of 10 days during the periovulatory period), 5 females were caught and dissected. From sampled organs, the gonado-, hepato- and viscerosomatic indexes (GSI, HSI, VSI) were calculated, oocyte diameters (OD) and the plasma levels of testosterone (T), 17P-estradiol (E2), 17,20P-dihydroxy-4-pregnen-3-one (17,2Op-P) and protein-phosphorus (PPP) were measured. After the sexual resting period observed from May to August (GSI \u3c 1 %, OD \u3c 200 urn, VSI = 4-6 %), oogenesis began in September when the water temperature decreased from 26.4 to 14.1 “C. The GSI increased progressively until mid March (15 %), then rapidly until spawning (25 %, OD = 850 urn) which occurred in April (14- 15 “C). The plasma levels of T, E,, 17,2Op-P and PPP were low during the sexual resting period. E, and PPP levels increased significantly at the onset of the oogenesis in September, then the E, level raised abruptly in November (3-4 ng mL-‘). In December, the T level increased rapidly to 15-20 ng . mL-’ The testosterone, E, and PPP levels remained very high until spawning, indicating the existence of active vitellogenesis. The highest HSI (2.1-2.2 %) recorded in winter confirmed this. During the periovulatory period, a peak of E, (4 ng . mL-‘) appeared, whereas T level diminished. In this study, 17,2Op-P levels remained low (0.2-0.6 ng mL-‘) and relatively constant. No 17,2Op-P peak was observed during the periovulatory period. Sampling at 10 day intervals was probably inadequate to specify the hormonal variations related to the final oocyte maturation and the ovulation. 0 Ifremer-Elsevier, Paris