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 species.

Abstract

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 microg/ml estradiol, and antibiotics) supplemented with either 0.5 microg/ml equine GH or 5 microg/ml equine LH. Porcine COCs were cultured in a basal medium (TCM199 with 570 microM cysteamine) supplemented with 0, 0.1, 0.5, or 1 microg/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 microg/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.