Glucose metabolism during bovine preimplantation development: analysis of gene expression in single oocytes and embryos.
Glucose metabolism of the bovine embryo is low during the first cleavages and increases sharply after the major resumption of the genome (8-16 cells). The mRNA level for genes involved in glucose metabolism was tested by RT-PCR on individual oocytes and embryos at different stages of development. These genes were: glucose transport GLUT-1, hexokinase (HK), glucose-6-phosphatase-dehydrogenase (G6PDH), and glucose-phosphate-isomerase (GPI); actin was used as a reference transcript. RT-PCR results revealed three types of oocytes or embryos: positive with a PCR signal for each transcript considered, nul with no signal for any transcript, and heterogeneous with a PCR signal for some transcripts and none for others. The number of nul and heterogeneous samples was higher for slow than for fast-cleaving embryos (81% vs. 36%), and the proportion of positive embryos increased significantly at the 16-cell and morula stages (P < 0.002), suggesting a correlation between mRNA content and developmental capacity. In positive embryos, GLUT-1 level was reduced by half during maturation and fertilization. Actin and hexokinase mRNA levels decreased during the first cleavages, but significantly increased at the 16-cell and morula stages, respectively. GPI transcript remained stable throughout development, whereas there was a significant rise for G6PDH at the 4-cell stage, perhaps due to a polyadenylation process. Finally, the absence or decrease in intensity of several transcripts at the blastocyst stage suggests suboptimal culture conditions.
Lequarre
A S
AS
Unité des Sciences Vétérinaires, Université Catholique de Louvain, Louvain-la-Neuve, Belgium.
Grisart
B
B
Moreau
B
B
Schuurbiers
N
N
Massip
A
A
Dessy
F
F
D000199:Actins / Q000235:genetics; D000818:Animals; D002417:Cattle; D004622:Embryo, Mammalian / Q000378:metabolism*; D047108:Embryonic Development; D005260:Female; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D005947:Glucose / Q000378:metabolism*; D051272:Glucose Transporter Type 1; D005956:Glucose-6-Phosphate Isomerase / Q000235:genetics; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics; D006593:Hexokinase / Q000235:genetics; D009004:Monosaccharide Transport Proteins / Q000235:genetics; D009865:Oocytes / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D014158:Transcription, Genetic
D000199:Actins; D051272:Glucose Transporter Type 1; D009004:Monosaccharide Transport Proteins; D005954:Glucosephosphate Dehydrogenase; D006593:Hexokinase; D005956:Glucose-6-Phosphate Isomerase; D005947:Glucose