Glucose metabolism assessed with 2-deoxyglucose and the effect of glutamate in subdivisions of rat hippocampal slices.
A new approach to the study of glucose phosphorylation in brain slices is described. It is based on timed incubation with nonradioactive 2-deoxyglucose (DG), after which the tissue levels of DG and 2-deoxyglucose-6-phosphate (DG6P) are measured separately with sensitive enzymatic methods applied to specific small subregions. The smallest samples had dry weights of approximately 0.5 microgram. Direct measurements in different regions of hippocampal slices showed that within 6 min after exposure to DG, the ratios of DG to glucose in the tissue were almost the same as in the incubation medium, which simplifies the calculation of glucose phosphorylation rates and increases their reliability. Data are given for ATP, phosphocreatine, sucrose space, and K+ in specific subregions of the slices. DG6P accumulation proceeded at a constant rate for at least 10 min, even when stimulated by 10 mM glutamate in the medium. The calculated control rate of glucose phosphorylation was 2 mmol/kg (dry weight)/min. In the presence of 10 mM glutamate it was twice as great. The response to 10 mM glutamate of different regions of the slice was not uniform, ranging from 164% of control values in the molecular layer of CA1 to 256% in the stratum radiatum of CA1. There was a profound fall in phosphocreatine levels (75%) in response to 10 mM glutamate despite a 2.4-fold increase in glucose phosphorylation. Even in the presence of 1 mM glutamate, the increase in glucose phosphorylation (50%) was not great enough to prevent a significant drop in phosphocreatine content.
Department of Molecular Biology and Pharmacology, Washington University School of Medicine, St. Louis, Missouri 63110.
D B
DB
McDougal
J G
JG
Carter
M E
ME
Pusateri
J K
JK
Manchester
O H
OH
Lowry
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001929:Brain Edema / Q000378:metabolism; D003847:Deoxyglucose / Q000378:metabolism*; D005260:Female; D005947:Glucose / Q000378:metabolism*; D019298:Glucose-6-Phosphate / Q000031:analogs & derivatives*; D005958:Glucosephosphates / Q000378:metabolism; D005971:Glutamates / Q000494:pharmacology*; D018698:Glutamic Acid; D006624:Hippocampus / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D010725:Phosphocreatine / Q000378:metabolism; D011188:Potassium / Q000378:metabolism; D051381:Rats
D005958:Glucosephosphates; D005971:Glutamates; D010725:Phosphocreatine; C015785:2-deoxyglucose-6-phosphate; D018698:Glutamic Acid; D019298:Glucose-6-Phosphate; D000255:Adenosine Triphosphate; D003847:Deoxyglucose; D005947:Glucose; D011188:Potassium