Activity of enzymes of energy metabolism in single human preimplantation embryos.
A method was developed to measure the activities of enzymes in extracts from single human preimplantation embryos. The method permits the analysis of two enzymes plus appropriate controls in an extract from a single embryo, and was used to investigate the control of energy metabolism during the development of human embryos from the two-cell to the blastocyst stage. Hexokinase (HK), 6-phosphofructokinase (PFK), pyruvate kinase (PK), fructose-1,6-diphosphate aldolase (ALD), glucose phosphate isomerase (GPI), lactate dehydrogenase (LDH), glucose-6-phosphate dehydrogenase (G6PDH) and 2-oxoglutarate dehydrogenase (ODH) were all detectable, whereas glycogen phosphorylase (GP) was not. The enzyme activities of ODH, PFK, LDH, PK, GPI and G6PDH, averaged over all stages of development from the two-cell to blastocyst stage (days 2-6 after insemination), were 3.5, 6.6, 15, 69, 73 and 87 times greater than HK, respectively. The activity of ALD was very similar to that of HK. The activities of ALD, GPI, PFK, PK and LDH showed no significant variation with stage of development, although the activity of GPI fell significantly from the four-eight cell to the eight-sixteen cell stage (P < 0.05). HK activity decreased from the two-eight cell to the eight-sixteen cell (P < 0.05), and increased significantly from the eight-sixteen cell to the blastocyst stage (P < 0.01). The overall relationship between hexokinase activity and stage approached significance (P = 0.059, one-way analysis of variance). The activity of G6PDH decreased significantly with development (P < 0.001, one way analysis of variance).(ABSTRACT TRUNCATED AT 250 WORDS)
Martin
K L
KL
Department of Biology, University of York, Heslington, UK.
Hardy
K
K
Winston
R M
RM
Leese
H J
HJ
D001755:Blastocyst / Q000201:enzymology*; D004734:Energy Metabolism / Q000502:physiology*; D005634:Fructose-Bisphosphate Aldolase / Q000378:metabolism; D005956:Glucose-6-Phosphate Isomerase / Q000378:metabolism; D005954:Glucosephosphate Dehydrogenase / Q000378:metabolism*; D006593:Hexokinase / Q000378:metabolism*; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D010732:Phosphofructokinase-1 / Q000378:metabolism; D006005:Phosphorylases / Q000378:metabolism*; D011770:Pyruvate Kinase / Q000378:metabolism
D007770:L-Lactate Dehydrogenase; D005954:Glucosephosphate Dehydrogenase; D006005:Phosphorylases; D006593:Hexokinase; D010732:Phosphofructokinase-1; D011770:Pyruvate Kinase; D005634:Fructose-Bisphosphate Aldolase; D005956:Glucose-6-Phosphate Isomerase