Simultaneous determination of local cerebral glucose utilization and blood flow by carbon-14 double-label autoradiography: method of procedure and validation studies in the rat

Journal Article · Sun Jun 01 04:00:00 EDT 1986 · J. Cereb. Blood Flow Metab.; (United States)

DOI:https://doi.org/10.1038/jcbfm.1986.49· OSTI ID:5272935

Ginsberg, M D; Smith, D W; Wachtel, M S; Gonzalez-Carvajal, M; Busto, R

Validation studies were undertaken to establish a computer-assisted double-label autoradiographic strategy employing (/sup 14/C)2-deoxyglucose ((/sup 14/C)2DG) and (/sup 14/C)iodoantipyrine ((/sup 14/C)IAP) to measure local CMRglu (LCMRglu) and CBF (LCBF). An organic solvent was used to extract the majority of IAP between first and second film exposures. In contrast to previously published data, all solvents tested produced partial losses of 2DG from tissue, and all allowed 2-6% of IAP to persist even after 5-day washes. Technical-grade chloroform permitted equal retention of unmetabolized and metabolized 2DG. A linear model was established, which was insensitive to the changes in tissue self-absorption that were shown to occur with chloroform extraction. Propagated error in computing tissue (/sup 14/C)2DG and (/sup 14/C)IAP was reduced by maximizing IAP extraction (by longer solvent wash times) and by administering 2.5 times as much IAP as 2DG. Fractional 2DG retention was measured in single-label 2DG sections placed on the films, and fractional IAP retention was evaluated by an optimization procedure. With this strategy, double-label values for LCMRglu and LCBF in anesthetized rats agreed with values obtained in matched single-label series to within 5%. The coefficients of variation for the double- and single-label LCMRglu data were virtually identical, whereas the coefficient of variation for double-label LCBF was 1.8 times that of single-label LCBF. The double-label strategy permitted pixel-by-pixel measurement and video display of the LCMRglu/LCBF ratio; the mean value among structures was 0.472 mumol/ml. With proper attention to methodological detail, this double-label strategy shows great promise for routine laboratory application.

• Research Organization: Univ. of Miami School of Medicine, FL
• OSTI ID: 5272935
• Journal Information: J. Cereb. Blood Flow Metab.; (United States), Journal Name: J. Cereb. Blood Flow Metab.; (United States) Vol. 3; ISSN JCBMD
• Country of Publication: United States
• Language: English

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59 BASIC BIOLOGICAL SCIENCES
ALDEHYDES
ANIMALS
ANTIPYRETICS
ANTIPYRINE
AUTORADIOGRAPHY
AZOLES
BLOOD FLOW
BODY
BRAIN
CARBOHYDRATES
CARBON 14 COMPOUNDS
CENTRAL NERVOUS SYSTEM
CENTRAL NERVOUS SYSTEM DEPRESSANTS
DOUBLE LABELLING
DRUGS
GLUCOSE
HETEROCYCLIC COMPOUNDS
HEXOSES
LABELLED COMPOUNDS
LABELLING
MAMMALS
MATHEMATICS
MEASURING METHODS
METABOLISM
MONOSACCHARIDES
NERVOUS SYSTEM
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANS
PYRAZOLES
PYRAZOLINES
RATS
REGRESSION ANALYSIS
RODENTS
SACCHARIDES
STATISTICS
VERTEBRATES