Genetic expression of hexokinase and glucose phosphate isomerase in late-stage mouse preimplantation embryos: transcription activities in glucose/phosphate-containing HTF and glucose/phosphate-free P1 media.
In mouse and human preimplantation development, pyruvate is consumed preferentially during early embryogenesis; however, during the morula and blastocyst stages, glucose is the preferred energy substrate. Studies have suggested that the glycolytic enzymes, hexokinase and glucose phosphate isomerase, are important enzymes in glucose metabolism during these later stages of human and mouse preimplantation development. In order to investigate the genetic activities of these enzymes in late-stage mouse embryos developing in vitro, we analysed hexokinase and glucose phosphate isomerase transcription activities by qualitative RNA assays using reverse transcriptase-nested polymerase chain reaction amplification of individual mouse morulae and early blastocysts incubated in glucose/phosphate-free preimplantation stage one (P1) medium and glucose/phosphate-containing human tubal fluid (HTF) medium. We observed an increased incidence of hexokinase transcripts in the population of blastocysts compared with morulae, and differences in transcript incidence between early blastocysts developing in HTF medium and in P1 medium. In contrast, glucose phosphate isomerase transcripts were consistantly present in all embryos analysed, and appear to be constitutively expressed during late-stage mouse embryogenesis. The different activity patterns of the two glycolytic genes may reflect different mechanisms of gene regulation or differential transcript stability during the later stages of mouse preimplantation development.
Johnson
M D
MD
Department of Gynecology and Obstetrics, Stanford University School of Medicine, CA 94305-5317, USA.
Batey
D W
DW
Behr
B
B
Barro
J
J
D000818:Animals; D001483:Base Sequence; D001755:Blastocyst / Q000201:enzymology* / Q000378:metabolism; D003470:Culture Media; D017931:DNA Primers / Q000235:genetics; D005260:Female; D018507:Gene Expression Regulation, Developmental; D005947:Glucose / Q000378:metabolism; D005956:Glucose-6-Phosphate Isomerase / Q000235:genetics*; D006593:Hexokinase / Q000235:genetics*; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D009028:Morula / Q000201:enzymology* / Q000378:metabolism; D010710:Phosphates; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011773:Pyruvates / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
D003470:Culture Media; D017931:DNA Primers; D010710:Phosphates; D011773:Pyruvates; D012333:RNA, Messenger; D006593:Hexokinase; D005956:Glucose-6-Phosphate Isomerase; D005947:Glucose