A bioluminescence technique for quantitative and structure-associated imaging of pyruvate.
A novel bioluminescence assay has been developed for measuring pyruvate within sections of snap-frozen tissue in a quantitative manner as well as with a spatial resolution on a microscopical level. The assay was verified via HPLC and two independent photometric tests. The novel assay makes it possible to determine pyruvate concentrations in cryosections in the range of 0-5.0 micromol/g tissue (dry weight). Based on the analysis of samples of given pyruvate concentrations, the assay exhibits a recovery with a deviation < or =15%. The minimal detectable amount was 0.02 pmol based on a 20 microm thick tissue section with an area of 1 cm(2). Combination of the already established imaging bioluminescence techniques for ATP, glucose, and lactate with the novel pyruvate assay allows for a comprehensive characterization of the metabolic profile of individual tumors. As the redox state of cancer cells can be critical for the efficiency of irradiation and a number of chemotherapeutics, and as pyruvate and lactate are known to have radical scavenger functions, we hypothesize that the novel bioluminescence assay may be used for measuring the pretherapeutic lactate-to-pyruvate ratio which may predict the radiosensitivity of individual malignancies.
Sattler
Ulrike G A
UG
Institute of Physiology and Pathophysiology, University of Mainz, Mainz, Germany.
Walenta
Stefan
S
Mueller-Klieser
Wolfgang
W
D001681:Biological Assay / Q000379:methods; D014408:Biomarkers, Tumor / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D003952:Diagnostic Imaging / Q000379:methods; D005629:Frozen Sections; D006019:Glycolysis / Q000502:physiology*; D006258:Head and Neck Neoplasms / Q000378:metabolism; D006801:Humans; D019344:Lactic Acid / Q000032:analysis / Q000378:metabolism; D008163:Luminescent Measurements / Q000379:methods*; D008164:Luminescent Proteins; D009132:Muscles / Q000378:metabolism*; D009135:Muscular Diseases / Q000235:genetics / Q000378:metabolism; D018307:Neoplasms, Squamous Cell / Q000378:metabolism; D010084:Oxidation-Reduction; D019289:Pyruvic Acid / Q000032:analysis* / Q000378:metabolism
D014408:Biomarkers, Tumor; D008164:Luminescent Proteins; D019344:Lactic Acid; D019289:Pyruvic Acid