WCRB2014 POSTER PRESENTATIONS (1) (335 abstracts)
Maturation conditions do not affect Myst1, Hat1, and Sirt1 mRNA abundance in horse oocytes
Irene Tessaro 1 , Federica Franciosi 2 , Pascal Papillier 3 , Ghylene Goudet 3 , Rozenn Dalbiès-Tran 3 , Cècile Douet 3 , Fabrice Reigner 3 , Stefan Deleuze 4 , Valentina Lodde 1 & Alberto M. Luciano 1
1University of Milan, Milan, Italy; 2University of California, San Francisco, San Francisco, California, USA; 3INRA, UMR85 Physiologie de la Reproduction et des Comportements, Nouzilly, France; 4Université de Liège, Liege, Belgium.
Introduction: We recently demonstrated that in vitro maturation (IVM) is associated with defects in histone H4 lysine 16 (H4K16) acetylation in horse oocytes, together with a higher incidence of spindle anomalies and aneuploidy. In the present study we investigated whether maturation conditions can alter the abundance of transcripts involved in histone acetylation and deacetylation. The specific H4K16 acetyl-transferase Myst1, the general acetyl-transferase Hat1 and the specific H4K16 deacetylase Sirt1 were examined in GV-stage oocytes and after IVM.
Methods: Follicle growth was followed by daily ultrasound scanning in adult mares. When a follicle >33 mm emerged, hCG was injected. After 35 h the IVM oocyte was collected by ultrasound-guided transvaginal aspiration. For IVM, oocytes were collected from follicles 5 to 25 mm and cultured for 28 h with EGF and serum. GV-stage oocytes and follicular cell samples were also collected from follicles 5 to 25 mm. Samples were retro-transcribed using random hexamers and analyzed by real-time Q-PCR. Gapdh and Renilla Luciferase served as internal and external housekeeping respectively.
Results and discussion: Our study reports for the first time the expression of Hat1, Myst1, and Sirt1 in horse oocytes and follicular cells. Hat1, Myst1, and Sirt1 were expressed at the same extent in GV-stage oocytes and after maturation, independently from the maturation conditions. Our findings show that the transcript abundance of Hat1, Myst1, and Sirt1 does not decrease during horse oocyte maturation, strongly suggesting that IVM does not promote/accelerate the degradation of transcripts, but rather affects H4K16 acetylation machinery at translational or post-translational level (L’Oreal 2012).
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ISSN 2052-1472 (online)
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