Now showing items 1-4 of 4

    • Article

      Gonadal response of juvenile protogynous grouper (Epinephelus fuscoguttatus) to long term recombinant follicle-stimulating hormone administration 

      P Palma, J Nocillado, J Superio, EGdJ Ayson, F Ayson, I Bar & A Elizur - Biology of Reproduction, 2019 - Oxford University Press
      The role of follicle stimulating hormone (FSH) in the gonadal development of protogynous hermaphroditic grouper (E. fuscoguttatus) was investigated. Recombinant giant grouper (E. lanceolatus) FSH (rggFSH) was produced in yeast. Its receptor binding capacity and steroidogenic potency were confirmed in vitro. Weekly injections of rggFSH to juvenile tiger grouper for 8 weeks (100 μg/kg body weight, BW) resulted in significantly larger and more advanced oocytes (cortical alveolar stage vs. primary growth stage in control). Sustained treatment with rggFSH (20 to 38 weeks at 200 μg/kg BW) resulted in significant reduction in gonad size, degeneration of oocytes and proliferation of spermatogonial cells, indicative of female to male sex change. Gene expression analysis showed that, while initiating female to male sex change, the rggFSH significantly suppressed the steroidogenic genes cyp11b, cyp19a1a and foxl2 which restrained the endogenous production of sex steroid hormones thus prevented the differentiation of spermatogonial cells. Expression profile of sex markers dmrt1, amh, figla and bmp15 suggests that the observed sex change was restricted at the initiation stage. Based on these results, we propose that the process of female to male sex change in the protogynous grouper is initiated by FSH, rather than sex steroids and likely involves steroid-independent pathway. The cortical alveolar stage in oocyte development is the critical point after which FSH-induced sex change is possible in grouper.
    • Article

      Induction of gonadal development in protogynous grouper with orally delivered FSH DNA 

      P Palma, J Nocillado, J Superio, EG de Jesus-Ayson, F Ayson, A Takemura, MW Lu & A Elizur - Marine Biotechnology, 2019 - Springer
      The availability of sexually mature fish often dictates the success of its captive breeding. In this study, we induced reproductive development in juvenile protogynous tiger grouper through oral administration of a plasmid (p) containing an engineered follicle-stimulating hormone (FSH). An expression construct (pcDNA3.1) was designed to express a single-chain FSH consisting of giant grouper FSH β-subunit and glycoprotein subunit-α (CGα), linked by the carboxy-terminal peptide (CTP) sequence from the human chorionic gonadotropin (hCG). Single oral delivery of pFSH encapsulated in liposome and chitosan to tiger grouper yielded a significant increase in plasma FSH protein level after 4 days. Weekly pFSH feeding of juvenile tiger groupers for 8 weeks stimulated ovarian development as indicated by a significant increase in oocyte diameter and progression of oocytes to cortical alveolar stage. As the pFSH treatment progressed from 20 to 38 weeks, female to male sex change was initiated, characterized by oocyte regression, proliferation of spermatogonial cells, and occurrence of spermatogenic cysts. It was also associated with significantly lower mRNA expression of steroidogenic genes (cyp11b, cyp19a1a, and foxl2) and basal plasma levels of sex steroid hormones 17β-estradiol (E2), testosterone (T), and 11-ketotestosterone (11KT). Results suggest that pFSH stimulates ovarian development up to cortical alveolar stage and then initiates sex change in tiger grouper. These findings significantly contribute to our knowledge on the role of FSH in the development of protogynous hermaphroditic fish. This study is the first to demonstrate induction of reproductive development in fish through oral delivery of plasmid gonadotropin.
    • Article

      Insights on the seasonal variations of reproductive features in the eastern Atlantic bluefin tuna 

      O Carnevali, F Maradonna, A Sagrati, M Candelma, F Lombardo, P Pignalosa, E Bonfanti, J Nocillado, P Palma, G Gioacchini & A Elizur - General and Comparative Endocrinology, 2019 - Elsevier
      The Atlantic Bluefin Tuna (ABFT, Thunnus thynnus) is one of the most intensely exploited fisheries resources in the world. In spite of the years of studies on ABFT, basic aspects of its reproductive biology remain uncertain. To gain insight regarding the seasonal changes of the reproductive characteristics of the eastern stock of ABFT, blood and tissue samples were collected from mature specimens caught in the Mediterranean basin during the reproductive (May-June) and non-reproductive season (Oct-Nov). Histological analysis of the gonads of May-June samples indicated that there were females which were actively spawning (contained post-ovulatory follicles) and females that were not actively spawning that had previtellogenic and fully vitellogenic oocytes. In males, testis were at early or late stage of spermatogenesis during the reproductive season. In Oct-Nov, ovaries contained mostly previtellogenic oocytes as well as β and α atretic follicles while the testis predominantly contained spermatogonia and few cysts with spermatocytes and spermatozoa. Gonadosomatic index (GSI) in females was highest among the actively spawning individuals while in males GSI was higher in early and late spermatogenic individuals compared to those that were spent. Plasma sex steroids levels varied with the reproductive season. In females, estradiol (E2), was higher in May-June while testosterone (T) and progesterone (P) did not vary. In males, E2 and T were higher in May-June while P levels were similar at the two sampling points. Circulating follicle stimulating hormone (FSH) was higher in Oct-Nov than in May-June both in males and females. Vitellogenin (VTG) was detected in plasma from both males and females during the reproductive season with levels in females significantly higher than in males. VTG was undetected in Oct-Nov samples. Since choriogenesis is an important event during follicle growth, the expression of three genes involved in vitelline envelope formation and hardening was measured and results showed significantly higher levels in ovaries in fish caught in May-June with respect to those sampled in Oct-Nov. In addition, a set of genes encoding for ion channels that are responsible for oocyte hydration and buoyancy, as well as sperm viability, were characterized at the two time points, and these were found to be more highly expressed in females during the reproductive season. Finally, the expression level of three mRNAs encoding for different lipid-binding proteins was analyzed with significantly higher levels detected in males, suggesting sex-specific expression. Our findings provide additional information on the reproductive biology of ABFT, particularly on biomarkers for the assessment of the state of maturation of the gonad, highlighting gender-specific signals and seasonal differences.
    • Article

      Reproductive development of the threatened giant grouper Epinephelus lanceolatus 

      The giant grouper is presumed to follow the reproductive pattern of most Epinephelus species, characterized by protogynous hermaphroditism wherein male maturation is attained through sex reversal of a functional female. This hypothesis, however, has not been verified due to lack of biological data. The present study addresses this gap by investigating the reproductive development of giant groupers from juvenile stage through sexual maturity. Gonad histological analysis of hatchery-bred juvenile giant grouper from Queensland, Australia (0.8–5.2 kg, n = 43) have shown earliest occurrence of primary oocytes (i.e. ovarian differentiation) in 47.8 cm and 2.5 kg fish. Monitoring of sexual maturity by gonadal biopsy was performed in a stock of wild-caught giant groupers (2–52 kg) held in sea cages in the Philippines and Vietnam from 2015 to 2017. Onset of female sexual maturity was at 96.9 ± 1.6 cm and 23.5 ± 1.5 kg in the Philippines, and 103.0 ± 4.1 cm and 33.5 ± 2.5 kg in Vietnam. In both locations, development of primary males was observed wherein fish produced milt (or spermiated) without passing through a functional female phase. The ratio of primary males to females in both locations was about 1:2. Size at maturity of primary males is 86.5 ± 4.8 cm and 17.1 ± 2.1 kg in the Philippines, and 97.3 ± 1.3 cm and 34.3 ± 0.9 kg in Vietnam. To aid in the monitoring of female maturation, we developed a non-invasive method based on immunoassay of vitellogenin in skin mucus and this was shown to be effective in detecting female maturation 9 ± 2 months prior to first observation of oocytes through gonadal biopsy. Our findings suggest that giant grouper is a diandric protogynous hermaphrodite. This study provides novel information on the reproductive biology of giant grouper, an economically important and vulnerable species.