Expression pattern of glucose metabolism genes correlate with development rate of buffalo oocytes and embryos in vitro under low oxygen condition

Abstract

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 % O 2 .Results Oocyte maturation and blastocyst formation rates were significantly higher at 5 % O 2 as compared to 20 % O 2 (P<0.05). The expression pattern of glycolytic genes (HK, PFK and G6PDH) indicated that oocytes and embryos under 5 % O 2 tend to follow anaerobic glycolysis and pentose phosphate pathways to support optimum embryo development. Under 20 % O 2 , oocytes and embryos had high expression of PDH indicating higher oxidative phosphorylation. Further, less G6PDH expression at 20 % O 2 was indicative of lower pentose phosphate activity. Higher expression of LDH was observed in oocytes and embryos under 20 % O 2 indicating sub-optimal culture conditions. High Glut1 activity was observed in the oocytes / embryos at 5 % O 2 , 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.

Ginsenoside Rg1 Improves <i>In vitro</i>-produced Embryo Quality by Increasing Glucose Uptake in Porcine Blastocysts

“…The results from this study show that, compared to positive and negative controls, 20 μg/mL ginsenoside increased GLUT1 expression. Because high levels of GLUT1 correlate with increased glucose uptake ( Kumar et al, 2015 ), these data indicate that glucose uptake was increased in ginsenoside-treated embryos. Ginsenoside stimulates glucose uptake through two pathways ( Shang et al, 2008 ; Lee et al, 2010b ), including the insulin signaling pathway by promoting the activation of both GLUT1 and GLUT4 ( Shang et al, 2008 ).…”

The Metabolomic Profile of Spent Culture Media from Day-3 Human Embryos Cultured under Low Oxygen Tension

“…Despite of the existence of evidences showing that the environmental stress created by using a higher oxygen tension during embryo culture may alter the metabolic activity of pre-implantation embryos [ 4 , 41 ], the multivariate statistics we used in our analysis showed that cleavage-stage human embryos grown under both types of oxygen concentration culture conditions left a similar metabolic fingerprint. We failed to observe any change in the net depletion or release of relevant analytes, such as glucose, certain amino acids, fatty acids and glycerophospholipids by human cleavage-stage embryos under either type of culture conditions.…”

Effects of glucose metabolism pathways on nuclear and cytoplasmic maturation of pig oocytes

“…Furthermore, studies on ETC in oocytes of different species are few [35][36][37] , and the role of NDUFV1, the first complex in ETC that accepts electrons from NADH and is critical for ATP generation, has not been observed in oocytes. In addition, although expression of different LDH isozymes were observed in oocytes and/or CCs of cattle 38 , human 39 , mouse 40 and buffalo 41 , their effects on oocyte maturation had not been carefully analyzed, and particularly, the effects had not been differentiated between DOs and CCs using such a powerful technique as RNAi.…”