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In vivo gene expression in granulosa cells during pig terminal follicular development

A Bonnet, INRA, Castanet-Tolosan, 31326, France
K Lê Cao, Genetique Animale, INRA, Castanet-Tolosan, France
M SanCristobal, Genetique Animale, INRA, Castanet-Tolosan, France
F Benne, Genetique Animale, INRA, Castanet-Tolosan, France
C Robert-Granié, Génétique Animale, INRA, Castanet, France
G Law-So, Genetique Animale, INRA, Castanet-Tolosan, France
S Fabre, PHASE, INRA, Tours, France
P Besse, Institut de mathematiques, Université Paul Sabatier, Toulouse, France
E De Billy, Cancer Research UK Center therapeutics, Institute of Cancer research, Sutton Surrey, United Kingdom
H Quesnel, Animal Physiology and Livestock Production Systems, INRA, Saint Gilles, France
F Hatey, Genetique Animale, INRA, Castanet-Tolosan, France
G Tosser-Klopp, Genetique Animale, INRA, Castanet-Tolosan, France

Correspondence: Agnès Bonnet, Email: abonnet{at}toulouse.inra.fr

Abstract

Ovarian antral follicular development is clearly dependent on pituitary gonadotropins FSH and LH. Although the endocrine mechanism that controls ovarian folliculogenesis leading to ovulation is quite well understood, the detailed mechanisms and molecular determinants in the different follicular compartments remain to be clarified. The aim of this study was to identify the genes differentially expressed in pig granulosa cells along the terminal ovarian follicle growth, to gain a comprehensive view of these molecular mechanisms.

First, we developed a specific micro-array using cDNAs from SSH libraries (345 contigs) obtained by comparison of 3 follicle size classes: small, medium and large antral healthy follicles. In a second step, a transcriptomic analysis using cDNA probes from these 3 follicle classes identified 79 differentially expressed transcripts along the terminal follicular growth and 26 predictive genes of size classes. The differential expression of 18 genes has been controlled using real time PCR experiments validating the micro-array analysis.

Finally, the integration of the data using Ingenuity Pathways Analysis identified five gene networks providing descriptive elements of the terminal follicular development. Specifically, we observed: 1) the down-expression of ribosomal protein genes, 2) genes involved in lipid metabolism and 3) the down-expression of cell morphology and ion binding genes.

In conclusion this study gives new insight into the gene expression during pig terminal follicular growth in vivo and suggested, in particular, a morphological change in pig granulosa cells accompanying terminal follicular growth.