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Testicular organ culture for the study of the retinoic acid signaling pathway in sea bass spermatogenesis

AuthorsMedina, Paula ; Gómez, Ana ; Zanuy, Silvia ; Blázquez, Mercedes
Issue DateJul-2013
Citation9th Congress of the Asociación Ibérica de Endocrinología Comparada. Final Program - Abstracts Book: 54 (2013)
AbstractThe first stages of spermatogenesis are characterized by the proliferation of spermatogonia and their subsequent entry into meiosis. The initiation of meiosis can thus be considered as the signal that triggers the onset of spermatogenesis. The role that the retinoic acid (RA) metabolic pathway has in the induction of meiosis has been demonstrated in humans, rodents, and amphibians, but studies in fish are still very scarce. The present work was aimed at the study of the gene expression that some key players of this pathway may have in the onset of meiosis in the European sea bass (Dicentrachus labrax). With this purpose, one year-old sea bass males were sacrificed and testis carefully extracted. The gonads where separated and immediately immersed in SBR buffer, washed, and finally placed on top of a nitrocellulose filter located on agarose cylinders previously soaked in supplemented L-15 media. Testicular explants were cultured during 48-72h in presence of DEAB, a RA synthesis inhibitor that exerts its action upon ALDH, an enzyme that oxidizes retinal into RA. In another set of experiments, explants were cultured during 48-72h in excess of all-trans RA. In both cases, the compounds were dissolved in DMSO and the possible effects of the solvent on the experiment were also checked. The study showed differences in expression levels of several genes involved in various steps of the RA signaling pathway. Moreover, DMSO can be used as a suitable carrier for this experimental set up since it had no effect on gene expression levels
Description9th Congress of the Asociación Ibérica de Endocrinología Comparada (AIEC), 13-14 July 2013, Barcelona, Spain
Appears in Collections:(IATS) Comunicaciones congresos
(ICM) Comunicaciones congresos
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