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Mol. Hum. Reprod. Advance Access originally published online on October 27, 2006
Molecular Human Reproduction 2007 13(1):3-9; doi:10.1093/molehr/gal089
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© The Author 2006. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions@oxfordjournals.org

Multiparameter assessment of mouse oogenesis during follicular growth in vitro

A. Pesty1,2,3,7, F. Miyara4, P. Debey4,5, B. Lefevre1,2,3 and C. Poirot6

1INSERM, 2Univ Paris, 3CEA, DRR/SEGG/LGAG, Fontenay-aux-Roses, 4INRA, Jouy-en-Josas, 5MHNH and 6AP, La Pitié Salpêtrière, UF Biologie de la Reproduction, Paris, France

7 To whom correspondence should be addressed : Arlette PESTY, INSERM, U-566, CEA, Batiment 05, 18 route du Panorama, Fontenay-aux-Roses, F-92260 France. E-mail: arlette.pesty{at}cea.fr


  Abstract

Comparison of oocyte development within the follicle in vitro and in vivo has a major impact on research into ovarian physiology and clinical practice. Despite major differences in ovarian physiology between rodents and humans, mice provide a useful model for studies of the endocrine and paracrine mechanisms controlling follicular development. In this study, early preantral follicles were isolated from 12-day-old mice and cultured individually in microdrops under oil during 6, 9 or 12 days. Taking into account previous observations, several oocyte criteria (diameter, chromatin configuration, transcriptional activity, intracytoplasmic calcium signalling and ability to undergo meiosis) were assessed to check that the development pattern of oocytes during follicle growth in vitro was similar to that already observed for oocytes developing in vivo, and that they reached the fertilizable oocyte stage. Results indicate that, during the 12-day-culture period, the oocytes grew until 74.3 ± 4.2 µm, they became transcriptionally quiescent with a surrounded nucleolus (SN) chromatin organization, 50% of them exhibited regular calcium signals and 73.4% of them resumed meiosis. These data demonstrate that the protocol used generates oocytes with characteristics similar to oocytes allowed to mature fully in vivo and that it could be useful to set up the experimental culture of human ovarian follicles.

Key words: calcium/chromatin/folliculogenesis/transcription


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