Inhibitors of glycolytic metabolism affect neurulation-staged mouse conceptuses in vitro.
In order to evaluate the apparent discordance between altered glucose metabolism and embryonic energy production, the effects of inhibitors of glucose utilization on morphological development and biochemical changes in mouse embryos in culture were evaluated. Day 9 ICR mouse conceptuses having 3-6 pairs of somites were prepared for culture as previously described. 2-Deoxyglucose (2DG) produced a concentration-dependent effect on development. A 25 microM 2DG concentration did not induce neural tube closure defects (NTDs) but 100 microM, 100% of embryos exhibited this defect. A 17% reduction in the rate of lactate production by the conceptus was produced by a 24-hr exposure period to 100 microM 2DG. Iodoacetate, which inhibits glyceraldehyde-3-phosphate dehydrogenase in adult tissues, produced high rates of NTDs at concentrations > or = 2.5 microM. Following a 24 hour exposure to iodoacetate, lactate production was inhibited at 10 and 25 microM. The effects of 2DG on embryonic ATP content were assessed to test the hypothesis that effects on glucose utilization would effect embryonic ATP content. Despite using 2DG concentrations that alter development and inhibit glycolysis, there were no effects on whole embryo or visceral yolk sac (VYS) ATP content. However, when the embryo was divided into regions, there was a specific reduction in ATP content in the head following a 24-hr exposure period. No effect of 2DG on head ATP content was produced after 12 hr of exposure. To determine if there were region specific differences in 2DG uptake and distribution that could account for the differential effects of 2DG on ATP content, 14C-2DG accumulation in different regions of the embryo and VYS was determined over the 24-hr culture period. The uptake of 2DG was dependent on the medium 2DG concentration and suggested a higher accumulation in regions with decreased ATP. However, when the uptake was monitored for a 1-hr period after a 24-hr exposure, there was no region specific differences in 2DG uptake. These studies further document the adverse developmental effects of inhibitors of glucose utilization during the early stage of neurulation. The biochemical mechanism for induction of these defects is unclear, but an effect on ATP content does not appear to be solely responsible for the dysmorphogenesis.
Hunter
E S
ES
Systems Toxicity Branch, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina 27709, USA.
Tugman
J A
JA
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D000963:Antimetabolites / Q000633:toxicity*; D003847:Deoxyglucose / Q000633:toxicity*; D004622:Embryo, Mammalian / Q000378:metabolism*; D005260:Female; D006019:Glycolysis / Q000187:drug effects*; D007461:Iodoacetates / Q000633:toxicity*; D019807:Iodoacetic Acid; D007773:Lactates / Q000096:biosynthesis; D019344:Lactic Acid; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D009420:Nervous System / Q000196:embryology* / Q000378:metabolism; D009436:Neural Tube Defects / Q000139:chemically induced*; D009924:Organ Culture Techniques; D011247:Pregnancy; D014018:Tissue Distribution
D000963:Antimetabolites; D007461:Iodoacetates; D007773:Lactates; D019344:Lactic Acid; D000255:Adenosine Triphosphate; D003847:Deoxyglucose; D019807:Iodoacetic Acid