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Master Thesis

Étude de l'impact de trois conditions d'élevage du poisson zèbre et mise au point d'une méthode de congélation de sa semence dans l'optique de réaliser une analyse comparative entre le niveau de méthylation de l'ADN des spermatozoïdes de poisson zèbre et d'étalon après congélation/décongélation

Abstract : DNA methylation is an epigenetic mark which participates in the regulation of gene expression. It is suggested that the methylation profile of spermatozoa is an important factor for early embryonic development. The purpose of this study was an evaluation of the impact of cryopreservation on sperm DNA methylation. We have selected two species with very different sperm nuclear organization, the zebrafish (Danio rerio) and the stallion. Zebrafish produce a very low quantity of gametes. In a first step, we sought to improve gamete production by way of the fish diet. We have shown that the Tétramin® + Gemma Wean® diet improved growth and favored gametes release. Gametes quality was equivalent to that obtained with a diet supplemented with live brine shrimps. In a second step, we tried to optimize the method of zebrafish sperm cryopreservation. The optimal cryopreservation method in our conditions was to use a cryoprotectant containing 8% of methanol and 63mM of sucrose at a cooling rate of -10°C/min. After cryopreservation, the level of DNA methylation of fish spermatozoa was altered completely randomly. Some sperm showed hypermethylation, other hypomethylation. No methylation data were obtained on stallion sperm, due to methodological difficulties whose origin was sought.
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Master Thesis
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https://dumas.ccsd.cnrs.fr/dumas-00941937
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Submitted on : Tuesday, February 4, 2014 - 3:00:09 PM
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Pierre Milon. Étude de l'impact de trois conditions d'élevage du poisson zèbre et mise au point d'une méthode de congélation de sa semence dans l'optique de réaliser une analyse comparative entre le niveau de méthylation de l'ADN des spermatozoïdes de poisson zèbre et d'étalon après congélation/décongélation. Sciences agricoles. 2013. ⟨dumas-00941937⟩

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