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This Article Full Text Full Text (PDF) All Versions of this Article: 69/3/1013    most recent biolreprod.103.015602v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Marchal, R. Articles by Goudet, G. Search for Related Content PubMed PubMed Citation Articles by Marchal, R. Articles by Goudet, G. Agricola Articles by Marchal, R. Articles by Goudet, G.

Effect of Growth Hormone (GH) on In Vitro Nuclear and Cytoplasmic Oocyte Maturation, Cumulus Expansion, Hyaluronan Synthases, and Connexins 32 and 43 Expression, and GH Receptor Messenger RNA Expression in Equine and Porcine Species¹

Unité de Physiologie de la Reproduction et des Comportements, Institut National de la Recherche Agronomique, 37380 Nouzilly, France

The aim of this study was to investigate the role of growth hormone (GH) on in vitro cumulus expansion and oocyte maturation in equine and porcine cumulus-oocyte complexes (COCs), and to approach its way of action. Equine COCs were cultured in a control medium (TCM199, 5 mg/ml BSA, 1 µg/ml estradiol, and antibiotics) supplemented with either 0.5 µg/ml equine GH or 5 µg/ml equine LH. Porcine COCs were cultured in a basal medium (TCM199 with 570 µM cysteamine) supplemented with 0, 0.1, 0.5, or 1 µg/ml porcine GH or in a control medium (basal medium with 10 ng/ml epidermal growth factor and 400 ng/ml FSH) supplemented with 0 or 0.5 µg/ml porcine GH. After culture, cumulus expansion and nuclear stage were assessed. The cytoplasmic maturation of porcine oocytes was evaluated by in vitro fertilization and development for 7 days. The modifications of the expression of proteins implicated in cumulus expansion were analyzed in equine COCs by SDS-PAGE with antibodies against connexins 32 and 43 and hyaluronan synthases (Has) 1, 2, and 3. The expression of GH receptor mRNA was studied in oocytes and cumulus cells of the two species using reverse transcription-polymerase chain reaction with specific primers. The addition of GH in maturation medium increased cumulus expansion in equine but not porcine COCs. It improved nuclear maturation in equine and porcine, but had no effect on porcine fertilization and embryo development. The GH receptor mRNA was detected in equine and porcine oocytes and cumulus cells. GH did not influence the expression of Has 1, Has 3, and connexin 43 in equine cumulus cells.

¹ Supported by grants from Les Haras Nationaux, France.

² Correspondence: Ghylène Goudet, Unité de Physiologie de la Reproduction et des Comportements, I.N.R.A., 37380 Nouzilly, France. FAX: 33 2 47 42 77 43; goudet{at}tours.inra.fr

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