Article (Scientific journals)
Analysis of Chromosome Segregation, Histone Acetylation, and Spindle Morphology in Horse Oocytes
Franciosi, Federica; Tessario, Irene; Dalbies-Tran, Rozenn et al.
2017In Journal of Visualized Experiments, 123 (e55242)
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Keywords :
developmental biology; oocyte maturation; meiosis; epigenetic; histone; ovum pick-up
Abstract :
[en] The field of assisted reproduction has been developed to treat infertility in women, companion animals, and endangered species. In the horse, assisted reproduction also allows for the production of embryos from high performers without interrupting their sports career and contributes to an increase in the number of foals from mares of high genetic value. The present manuscript describes the procedures used for collecting immature and mature oocytes from horse ovaries using ovum pick-up (OPU). These oocytes were then used to investigate the incidence of aneuploidy by adapting a protocol previously developed in mice. Specifically, the chromosomes and the centromeres of metaphase II (MII) oocytes were fluorescently labeled and counted on sequential focal plans after confocal laser microscope scanning. This analysis revealed a higher incidence in the aneuploidy rate when immature oocytes were collected from the follicles and matured in vitro compared to in vivo. Immunostaining for tubulin and the acetylated form of histone four at specific lysine residues also revealed differences in the morphology of the meiotic spindle and in the global pattern of histone acetylation. Finally, the expression of mRNAs coding for histone deacetylases (HDACs) and acetyl-transferases (HATs) was investigated by reverse transcription and quantitative-PCR (q-PCR). No differences in the relative expression of transcripts were observed between in vitro and in vivo matured oocytes. In agreement with a general silencing of the transcriptional activity during oocyte maturation, the analysis of the total transcript amount can only reveal mRNA stability or degradation. Therefore, these findings indicate that other translational and post-translational regulations might be affected. Overall, the present study describes an experimental approach to morphologically and biochemically characterize the horse oocyte, a cell type that is extremely challenging to study due to low sample availability. However, it can expand our knowledge on the reproductive biology and infertility in monovulatory species.
Disciplines :
Anatomy (cytology, histology, embryology...) & physiology
Author, co-author :
Franciosi, Federica;  Università degli Studi di Milano > Department of Health > animal science and food safety
Tessario, Irene;  Università degli Studi di Milano > Department of Health > Animal science and food safety
Dalbies-Tran, Rozenn
Douet, Cecile;  Institut Scientifique de Recherche Agronomique - INRA > PRC
Reigner, Fabrice;  Institut Scientifique de Recherche Agronomique - INRA > UEPAO
Deleuze, Stefan  ;  Université de Liège > Dép. clinique des animaux de compagnie et des équidés (DCA) > Obstét. et path. de la reprod. des anim. de comp. et équidés
Papiller, Pascal;  Institut Scientifique de Recherche Agronomique - INRA > PRC
Maclea, Ileana;  University of agricultural science and veterinary medicine, Cluj-Napoca
Lodde, Valentina;  Università degli Studi di Milano > Department of health > Animal science and food safety
Luciano, Alberto;  Università degli Studi di Milano > Department of health > Animal science and food safety
Goudet, Ghylene;  Institut Scientifique de Recherche Agronomique - INRA > PRC
Language :
English
Title :
Analysis of Chromosome Segregation, Histone Acetylation, and Spindle Morphology in Horse Oocytes
Publication date :
2017
Journal title :
Journal of Visualized Experiments
eISSN :
1940-087X
Publisher :
MYJoVE Corporation, Boston, United States - Massachusetts
Volume :
123
Issue :
e55242
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 12 June 2017

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