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Rat Brain Free Glucose and Lactate Measurement by a Novel Method Using Bisecting Decapitation‐Extrusion and Enzyme Denaturation at Five Seconds
Author(s) -
Oldendorf William H.,
Stoller Barret E.
Publication year - 1991
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1991.tb08193.x
Subject(s) - denaturation (fissile materials) , sonication , chemistry , urea , enzyme , centrifugation , extrusion , chromatography , biochemistry , enzyme assay , biophysics , biology , materials science , nuclear chemistry , metallurgy
This study investigates a novel method as a means for animal decapitation with rapid brain removal and enzyme denaturation. Briefly, the rat head is simultaneously decapitated and bisected. Either half of the in situ brain is aspirated under —250 mm Hg pressure into a modified small plastic syringe and then extruded through a needle as a fine strand into a relatively large volume of 2 M urea at 95°C. After cooling, sonication, and centrifugation of the brain homogenate, the supernatant is measured enzymatically for brain free glucose and lactate concentration. Enzyme denaturation is effected within 4–6 s. The results are in good agreement with published values for glucose and lactate using other rapid enzyme inactivation techniques.