Rat brain free glucose and lactate measurement by a novel method using bisecting decapitation-extrusion and enzyme denaturation at five seconds.
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 degrees 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.
Oldendorf
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WH
Research Service, West Los Angeles Veterans Administration, California 90073.
Stoller
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BE
D000758:Anesthesia; D000818:Animals; D001921:Brain / Q000201:enzymology*; D001923:Brain Chemistry; D005947:Glucose / Q000032:analysis*; D006019:Glycolysis; D006257:Head; D007773:Lactates / Q000032:analysis*; D019344:Lactic Acid; D008297:Male; D010424:Pentobarbital; D011180:Postmortem Changes; D011489:Protein Denaturation; D051381:Rats; D011919:Rats, Inbred Strains; D013396:Suction / Q000295:instrumentation; D013997:Time Factors; D014508:Urea
D007773:Lactates; D019344:Lactic Acid; D014508:Urea; D010424:Pentobarbital; D005947:Glucose