Dépôt numérique

Establishment of an in vitro model to study epigenetic reprogramming in rat fetal male germ cells

Rwigemera, Arlette et Delbès, Géraldine . Establishment of an in vitro model to study epigenetic reprogramming in rat fetal male germ cells In: 9ème symposium annuel du Réseau Québécois en Reproduction, Novembre 2016, Québec.

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Epigenetic reprogramming is a critical step in male germ cells development that occurs during fetal life. It is characterized by the remodeling of different epigenetic marks such as DNA methylation (5mC) and post - translational modifications of histone H3. It was suggested that exposure to some environmental pollutants affect male fertility especially if it occurs during the phase of epigenetic reprogramming. The organ culture is a powerful system that reproduces the in vivo kinetics of testicular developmen t. We hypothesize that organ culture can reproduce the epigenetic reprogramming and therefore would be a good model to study the impact of pollutants on testis development. We aim to test if this model can reproduce the epigenetic reprogramming in gonocyte s by quantifying the ex vivo dynamic of epigenetic marks that are known to vary during that time in vivo. First, using immunofluorescence on testis explanted at four different stages of development, we showed that 5 histone modifications and 5mC have dynam ic profiles in rat gonocytes during that time. Second, we explanted testis at 16 dpc and cultured them with/without FBS. Without FBS, the immunofluorescence quantification results done after 48h or 96h of culture, show that the marks tested (5mC, H3K4me2, H3K4me3) follow similar patterns as the one observed in vivo. We then used the pyrosequencing technique to establish the kinetic of re - methylation of the paternally imprinted gene H19 in gonocytes in vivo and showed that that kinetic is reproduced in vitro . These results suggest that the organ culture can reproduce the process of epigenetic reprogramming.

Type de document: Document issu d'une conférence ou d'un atelier
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Centre: Centre INRS-Institut Armand Frappier
Date de dépôt: 01 mai 2018 15:38
Dernière modification: 19 oct. 2023 13:18
URI: https://espace.inrs.ca/id/eprint/5941

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