RESUMO
Efforts to establish an aquaculture industry for sablefish (Anoplopoma fimbria) are constrained by reproductive dysfunction in wild-caught fish and by lack of reproduction of F1 females. Toward a better understanding of the reproductive dysfunction of captive broodstock, full-length cDNAs encoding the sablefish gonadotropin subunits (fshb, lhb and cga) and their receptors (fshr and lhcgr) were cloned, sequenced and quantitative real-time PCR assays developed. Sablefish gonadotropin subunits display some unique features, such as two additional Cys residues in the N-terminal region of Fshb and a lack of potential N-glycosylation sites in Fshb and Lhb, whereas Fshr and Lhcgr possess conserved structural characteristics described in other vertebrates. Wild females captured in fall completed gametogenesis in captivity the next spawning season, whereas females captured three months earlier, during summer, failed to mature. Interestingly, these wild non-maturing females exhibited similar reproductive features as prepubertal F1 females, including low levels of pituitary gonadotropin and ovarian receptor mRNAs and plasma sex steroids, and ovarian follicles arrested at the perinucleolus stage. In conclusion, this study described the cloning, molecular characterization and development of qPCRs for sablefish gonadotropins and their receptors. Rearing conditions may impair vitellogenic growth of ovarian follicles in sablefish, compromising the reproductive success of broodstock.
Assuntos
Peixes/metabolismo , Gonadotropinas/metabolismo , Receptores da Gonadotropina/metabolismo , Animais , DNA Complementar/genética , Feminino , Peixes/genética , Gametogênese/genética , Gametogênese/fisiologia , Gonadotropinas/genética , Masculino , Folículo Ovariano/metabolismo , Receptores da Gonadotropina/genéticaRESUMO
Mechanisms regulating the normal progression of ovarian follicular growth versus onset of atresia in fishes are poorly understood. To gain a better understanding of these processes, we exposed immature female coho salmon (Oncorhynchus kisutch) to prolonged fasting to induce follicular atresia and monitored body growth, development of the ovarian follicles, changes in reproductive hormones, and transcripts for ovarian genes. Prolonged fasting reduced body and ovary weight and increased the appearance of atretic follicles relative to normally fed controls. Endocrine analyses showed that fasting reduced plasma insulin-like growth factor 1 (IGF1), estradiol-17ß (E2), and pituitary, but not plasma, levels of follicle-stimulating hormone (FSH). Transcripts for ovarian fsh receptor (fshr) and steroidogenesis-related genes, such as steroidogenic acute regulatory protein (star), 3ß-hydroxysteroid dehydrogenase (hsd3b), and P450 aromatase (cyp19a1a) were significantly lower in fasted fish. Ovarian expression of apoptosis-related genes, such as Fas-associated death domain (fadd), caspase 8 (casp8), caspase 3 (casp3), and caspase 9 (casp9) were significantly elevated in fasted fish compared to fed fish, indicating that apoptosis is involved in the process of atresia in this species. Interestingly, some genes such as fadd, casp8, casp3, and hsd3b, were differentially expressed prior to increases in the number of atretic follicles and reductions in hormone levels induced by fasting, and may therefore have potential as early indicators of atresia. Together these results suggest that prolonged nutritional stress may disrupt the reproductive system and induce follicular atresia in part via reductions in ovarian IGF and FSH signaling, and downstream effects on steroidogenesis-related genes and E2 production.