Quantitative on-line monitoring of hippocampus glucose and lactate metabolism in organotypic cultures using biosensor technology.
Quantitative glucose and lactate metabolism was assessed in continuously perfused organotypic hippocampal slices under control conditions and during exposure to glutamate and drugs that interfere with aerobic and anaerobic metabolism. On-line detection was possible with a system based on slow perfusion rates, a half-open (medium/air interface) tissue chamber and a flow injection analytic system equipped with biosensors for glucose and lactate. Under basal conditions about 50% of consumed glucose was converted to lactate in hippocampal slice cultures. Using medium containing lactate (5 mm) instead of glucose (5 mm) significant lactate uptake was observed, but this uptake was less than the net uptake of lactate equivalents in glucose-containing medium. Glucose deprivation experiments suggested lactate efflux from glycogen stores. The effects of drugs compromising or stimulating energy metabolism, i.e. 2-deoxyglucose, 3-nitropropionic acid, alpha-cyano-4-hydroxycinnamate, l-glutamate, d-asparate, ouabain and monensin, were tested in this flow system. The data show that maintaining Na+ and K+ gradients consumed much of the energy but do not support the hypothesis that l-glutamate stimulates glycolysis in hippocampal slice cultures.
Gramsbergen
Jan Bert
JB
Department of Anatomy and Neurobiology, SDU-Odense University, Odense, Denmark. jbgramsbergen@health.sdu.dk
Leegsma-Vogt
Gea
G
Venema
Kor
K
Noraberg
Jens
J
Korf
Jakob
J
D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D046508:Culture Techniques / Q000379:methods; D004734:Energy Metabolism / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005947:Glucose / Q000032:analysis / Q000378:metabolism* / Q000493:pharmacokinetics; D006624:Hippocampus / Q000187:drug effects / Q000378:metabolism*; D019344:Lactic Acid / Q000032:analysis / Q000378:metabolism*; D008985:Monensin / Q000494:pharmacology; D009498:Neurotoxins / Q000494:pharmacology; D009574:Nitro Compounds; D010042:Ouabain / Q000494:pharmacology; D011422:Propionates / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D000254:Sodium-Potassium-Exchanging ATPase / Q000187:drug effects
D004791:Enzyme Inhibitors; D009498:Neurotoxins; D009574:Nitro Compounds; D011422:Propionates; D019344:Lactic Acid; D010042:Ouabain; D008985:Monensin; D000254:Sodium-Potassium-Exchanging ATPase; D005947:Glucose; C015392:3-nitropropionic acid