Macromolecules Influence Cellular Competence and Expression Level of <i>IGFs</i> Genes in Bovine Oocytes In Vitro
Uğur Şen | Emre Şirin | Hasan Önder | Selçuk Özyürek | Magdalena Kolenda | Beata Sitkowska
In vitro maturation (IVM) of mammalian oocytes, which influences subsequent in vitro development of embryos, is affected by the macromolecule content in culture media for the success of oocyte maturation competence, in which the cytoplasmic and nuclear reprogramming events occur. The insulin-like growth factor family (<i>IGFs</i>) promotes the maturation of bovine oocytes and the expansion of cumulus cells and also inhibits apoptosis. This study was, therefore, designed to examine the effects of macromolecules (bovine serum albumin, BSA; fetal calf serum, FCS; and polyvinyl alcohol, PVA) on in vitro nuclear maturation, total cellular protein, glutathione peroxidase (GPx) enzyme activity, and the gene expression level of <i>IGF1</i>, <i>IGF2</i>, and their receptor in bovine oocytes. Oocytes obtained from bovine ovaries were cultured in bicarbonate-buffered medium 199 supplemented with 4 mg/mL BSA, 10% FCS, 1 mg/mL PVA, and without macromolecule supplement (control) during 22 h in the air with a humidified atmosphere and 5% CO<sub>2</sub> at 38.5 °C temperature. Supplementation of BSA and FCS increased (χ<sup>2</sup> = 9.84; <i>p</i> < 0.05) the percentages of oocytes that reached metaphase II compared to the control and PVA. The amount of protein per ml of cell extracts of oocytes matured in FCS supplemented culture media was higher (<i>p</i> < 0.05) than the oocytes in the PVA and control. The levels of GPx enzyme activity in cell extracts isolated from oocytes in each experimental group did not change over time, but the GPx enzyme activity in oocytes matured in PVA-supplemented culture media was lower (<i>p</i> < 0.05) than in oocytes in the other experimental groups. Transcript for the <i>IGF1</i> gene was not detected in all experimental groups, but the supplementation of BSA and FCS significantly elevated the transcript level of the <i>IGF2</i> gene. In addition, the maturation of oocytes with BSA-supplemented media increased the transcript level of the <i>IGF1R</i> gene, whereas the transcript level of the <i>IGF2R</i> gene was similar among macromolecule supplementation groups. The current study concluded that BSA and FCS could improve in vitro bovine oocyte development due to supporting nuclear maturation and increasing the total cellular protein content, GPx enzyme, and transcript activity.
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