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Specific recombinant gonadotropins induce for the first time in European eel (Anguilla anguilla) full spermiating fish from immature males

AuthorsPeñaranda, D. S.; Gallego, V.; Rozenfeld, C.; Vílchez, M.C.; Pérez, Luz; Gómez, Ana ; Giménez, Ignacio; Asturiano, Juan F.
Issue Date14-Nov-2016
Citation22nd International Conference of Zoology (2016)
AbstractIn teleosts, recombinant gonadotropins (rGTHs), both in vitro and in vivo, have been able to induce the steroidogenesis and gonad development, however the in vivo results have been variable. In vivo, specific rGTH stimulated estradiol (E2) production in female of species such as orange-spotted grouper (Epinehelus coioides) or rosy bitterling (Rhodeus ocellatus), achieving ovulation in the latter, in that study however, already sexually matured females were used. In males, rGTHs were also able to induce the androgenesis process e.g. in zebrafish (Danio rerio) and European sea bass (Dicentrarchus labrax). Generally, the hormonal treatment was not able to induce the spermiation in vivo, with the exception of goldfish (Carassius auratus) and European sea bass, where the treated fish were already sexually matured at the beginning of the study. In Japanese eel (Anguilla japonica), recombinant FSH stimulated the in vitro production of E2 and testosterone in oocytes, but only at vitellogenic. In vivo, rGTHs did not stimulate an increase in the gonadosomatic index in females, but some induction of vitellogenesis was observed. In males, rGTHs expressed by baculovirus, yeasts or Drosophila cell lines were able to induce androgenesis in vitro. A complete spermatogenesis, with the presence of some spermatozoa in the testis, was achieved with rGTHs expressed by baculovirus. In vivo, Japanese eel and goldfish rGTHs, expressed by baculovirus, stimulated a complete spermatogenesis in male eels, but spermiation has never been achieved. Therefore, in the current work, the aim was to check the functionality in vivo of specific rGTHs in European eel. European eel males (n=72; 100.1±1.9 g) from a local fish farm were distributed in four 150-L aquaria and progressively adapted to sea water and 20 ºC. Single-chain recombinant FSH and LH were obtained by transfection of a mammalian cell line with further partial purification and up-concentration (Rara Avis Biotec S.L.; Valencia, Spain). In a first experiment, males were submitted to five hormonal treatments: one group was treated during 9 weeks with constant doses of recombinant European eel follicle-stimulating hormone (aarFSH; 4 µg/fish), a second group received constant doses of recombinant European eel luteinizing hormone (aarLH; 2 µg/fish) also during 9 weeks, while other three groups were injected with different combinations of constant or changing doses of both aarGTHs. All treatments stimulated the androgen synthesis, but this increase was more pronounced in those fish treated with a combination of both aarGTHs. Unlike aarLH or aarFSH alone were able to induce spermiating males, the fish treated with constant doses of aarFSH and increasing ones of aarLH registered the best results, with 20% of motile spermatozoa, stimulating spermiation for first time in a teleost from immature fish. In order to improve the sperm quality, a second experiment was performed. Immature males received three constant aarFSH doses (2.8, 1.4 or 0.7 µg/fish) and increasing doses of aarLH (every 3 weeks; 1, 2, 6 µg/fish). All treatments induced spermiating males, however the best sperm quality (¿50% motile spermatozoa) was observed in males treated with the highest aarFSH dose. In conclusion, these new specific recombinant gonadotropins have demonstrated their capacity to induce the spermatogenesis and spermiation in vivo in a teleost fish, the European eel.
DescriptionTrabajo presentado en la 22nd International Conference of Zoology, celebrado en Okinawa (Japón), del 14 al 19 de noviembre de 2016
Appears in Collections:(IATS) Comunicaciones congresos
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