PURPOSE: This study evaluates the effect of low oxygen conditions (5 Vs 20%) on buffalo embryo development. Expression patterns of key glucose metabolism genes (HK, PFK, LDH, PDH, G6PDH and Glut1) were assessed in buffalo oocytes and embryos cultured at 5 and 20% oxygen and correlated with development rate. METHODS: Maturation rate was observed by determining MII stages by Aceto-orcein method and blastocyst formation was observed at 7 day post insemination (dpi). Expression levels of genes were determined by real time PCR in oocytes / embryos at 5 and 20% O2. RESULTS: Oocyte maturation and blastocyst formation rates were significantly higher at 5% O2 as compared to 20% O2 (P < 0.05). The expression pattern of glycolytic genes (HK, PFK and G6PDH) indicated that oocytes and embryos under 5% O2 tend to follow anaerobic glycolysis and pentose phosphate pathways to support optimum embryo development. Under 20% O2, oocytes and embryos had high expression of PDH indicating higher oxidative phosphorylation. Further, less G6PDH expression at 20% O2 was indicative of lower pentose phosphate activity. Higher expression of LDH was observed in oocytes and embryos under 20% O2 indicating sub-optimal culture conditions. High Glut1 activity was observed in the oocytes / embryos at 5% O2, indicative of high glucose uptake correlating with high expression of glycolytic genes. CONCLUSION: The expression patterns of glucose metabolism genes could be a valuable indicator of the development potential of oocytes and embryos. The study indicates the importance of reduced oxygen conditions for production of good quality embryos.
PURPOSE: This study evaluates the effect of low oxygen conditions (5 Vs 20%) on buffalo embryo development. Expression patterns of key glucose metabolism genes (HK, PFK, LDH, PDH, G6PDH and Glut1) were assessed in buffalo oocytes and embryos cultured at 5 and 20% oxygen and correlated with development rate. METHODS: Maturation rate was observed by determining MII stages by Aceto-orcein method and blastocyst formation was observed at 7 day post insemination (dpi). Expression levels of genes were determined by real time PCR in oocytes / embryos at 5 and 20% O2. RESULTS: Oocyte maturation and blastocyst formation rates were significantly higher at 5% O2 as compared to 20% O2 (P < 0.05). The expression pattern of glycolytic genes (HK, PFK and G6PDH) indicated that oocytes and embryos under 5% O2 tend to follow anaerobic glycolysis and pentose phosphate pathways to support optimum embryo development. Under 20% O2, oocytes and embryos had high expression of PDH indicating higher oxidative phosphorylation. Further, less G6PDH expression at 20% O2 was indicative of lower pentose phosphate activity. Higher expression of LDH was observed in oocytes and embryos under 20% O2 indicating sub-optimal culture conditions. High Glut1 activity was observed in the oocytes / embryos at 5% O2, indicative of high glucose uptake correlating with high expression of glycolytic genes. CONCLUSION: The expression patterns of glucose metabolism genes could be a valuable indicator of the development potential of oocytes and embryos. The study indicates the importance of reduced oxygen conditions for production of good quality embryos.
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