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Doctoral Thesis
Full name
Chayrra Chehade Gomes
Knowledge Area
Date of Defense
São Paulo, 2015
Borella, Maria Ines (President)
Gomes, Vicente
Jaeger, Ruy Gastaldoni
Lopes, Marilene Hohmuth
Whitton, Renata Guimarães Moreira
Title in Portuguese
Caracterização molecular dos GnRHs de Astyanax altiparanae (Garutti e Britski, 2000), seu efeito in vivo, e sua expressão temporal ao longo do estímulo reprodutivo.
Keywords in Portuguese
Indução à reprodução
Abstract in Portuguese
Sendo o GnRH a molécula capaz de iniciar a cascata hormonal reprodutiva, realizamos a clonagem, analisamos os efeitos da indução à reprodução e as funções dos GnRHs de A. altiparanae. Como resultados, obtivemos as sequências dos cDNAs do GnRH2 e GnRH3. Quando induzidos à reprodução, a desova foi às 20 h pós-estímulo (hpe) em fêmeas e às 16 hpe em machos, o aumento da expressão do mRNA de GnRH3 ocorreu às 8 hpe em fêmeas, e o aumento da expressão do mRNA de GnRH2 foi às 0 hpe em machos. Com relação ao efeito dos GnRHs, todos estimularam a expressão do mRNA de βLH, mas não de βFSH, e só o GnRH2 foi capaz de elevar o MIS e causar a desova. Como conclusão, temos que as sequências dos cDNAs dos GnRHs se mostraram conservadas; a indução à reprodução por redução do nível da água foi eficaz; em cativeiro, a espécie teve desenvolvimento assincrônico; o GnRH2 provavelmente está ligado ao comportamento reprodutivo, e o GnRH3 é a possível forma hipofisiotrópica. Por fim, só o GnRH2 desencadeou toda a cascata hormonal.
Title in English
Molecular characterization of GnRH of the Astyanax altiparanae (Garutti and Britski, 2000), its effect in vivo, and its temporal expression during the reproductive stimulus.
Keywords in English
Induction of reproduction
Abstract in English
The study on reproduction in fish has been acquired great importance in last years, mainly for the benefit of threatened species. During the reproductive process, the hypothalamic neurons synthesize and release GnRH that stimulates the pituitary cells to release FSH and LH, which, in turn, promote the gonadal maturation. In fact, the morphological changes in gonads are the result of the endocrine action of the reproductive axis, in which the GnRH is the key molecule to starting the reproductive axis control. Thus, the knowledge about the GnRH, as well as about the gonadal morphological changes in the spawning might contribute to effectiveness of reproduction. Therefore, in this work, with Astyanax altiparanae as a model, we made the molecular characterization of the GnRHs, and we analyzed the gonadal morphological changes during the reproductive stimulus. In addition, we evaluated the role of injected GnRHs in vivo. As results, we obtain the cDNA complete sequence of preproGnRH2 (612bp) and preproGnRH3 (407bp) of A. altiparanae. Regarding the induction of reproduction by water level drawdown, the released of gametes occurred at 20 hours after stimulus in female and at 16 hours after stimulus in males, the mRNA expression of GnRH3 increased at 8 hours after stimulus in female and the mRNA expression of GnRH2 increased at 0 hours in males. Regarding the effects of injected GnRH, all of them stimulated the βLH but not βFSH mRNA expression, and only the GnRH2 was able to rise the MIS and stimulate the released of gametes. We conclude that the cDNAs sequences of preproGnRH2 and preproGnRH3 were conserved, although there is a change in the amino acid at the position 8 of the GnRH3 decapeptide of A. altiparanae. Furthermore, the induced reproduction by water level drawdown was effective, and in captivity, the A. altiparanae has an asynchronous development with splitted spawning during the breeding season. The analysis of the animals submitted to the reproductive stimulus allowed us to suggest that in A. altiparanae, the GnRH2 probably has a role in sexual behavior and the GnRH3 possibly is the hypophysiotropic form. Finally, analyzing the GnRH effects, we observed that only the GnRH2 was able to start the entire reproductive hormonal cascade, leading the animal to spawning.
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