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Mol. Hum. Reprod. Advance Access published online on June 24, 2009
Molecular Human Reproduction, doi:10.1093/molehr/gap051
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© The Author 2009. 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

Quantification of oocyte-specific transcripts in follicle-enclosed oocytes during antral development and maturation in vitro

Flor Sánchez1,2, Tom Adriaenssens1, Sergio Romero1 and Johan Smitz1

1Follicle Biology Laboratory, Vrije Universiteit Brussel, 1090 Brussels, Belgium

2 Correspondence addresses: Fax. 32 2 477 50 60; Email: fsanchez{at}vub.ac.be

Oocyte cytoplasmic maturation is influenced by the quantity of synthesized RNA and proteins accumulated and stored during growth. Transcriptional repression and degradation of transcripts occur during oocyte nuclear maturation and prolonged transcriptional arrest might compromise RNA stores for early development. RNA quantification of key genes in oocytes might be valuable when setting-up in vitro cultures that lack the normal hormonal interplay found in vivo.

This study quantitates gene expression levels in relation to follicle culture time and time of oocyte maturation in a mouse model. RNA levels of Gdf-9, Bmp-15, Mater, Zar-1, Npm-2 and Fgf-8 were measured in germinal vesicle (GV) oocytes along fixed times during in vitro follicle development. For all genes the highest mRNA levels were detected in oocytes in the preantral follicle stage. Antrum formation was associated with a progressive shutdown in transcription in extended in-vitro culture leading to lower mRNA values than those of control in vivo preovulatory oocytes. In contrast to in vitro matured oocytes, the in vivo oocytes from 22 and 29 day-old prepubertal animals obtained after pregnant mare's serum globulin and hCG priming did not downregulate transcripts upon maturation stimulus except for Mater.

These findings show that oocyte gene expression patterns under in vitro conditions can, at certain times, mimic what is reported to occur under in vivo conditions. Moreover, they also show that meiotically competent oocytes kept in a prolonged transcriptionally inactive stage express altered levels of key transcripts compared to in vivo in both immature and mature oocytes.

Key Words: Oocyte-specific genes/gene expression/developmental competence/follicle culture/oocyte maturation

Submitted on March 3, 2009; resubmitted on June 11, 2009; accepted on June 19, 2009.


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