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Journal of Histochemistry and Cytochemistry, Vol. 45, 1217-1230, Copyright © 1997 by The Histochemical Society, Inc.

ARTICLE

In Situ Measurement of Glutamate Concentrations in the Periportal, Intermediate, and Pericentral Zones of Rat Liver

Willie J. C. Geertsa, Ard Jonkera,b, Louis Boonb, Alfred J. Meijerc, Rob Charlesa, Cornelis J. F. Van Noordenb, and Wouter H. Lamersa
a Academic Medical Centre, University of Amsterdam, Department of Anatomy and Embryology, Amsterdam, The Netherlands
b Department of Cell Biology and Histology, Amsterdam, The Netherlands
c Department of Biochemistry, Amsterdam, The Netherlands

Correspondence to: Wouter H. Lamers, Dept. of Anatomy and Embryology, Academic Medical Centre, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands.

We developed a quantitative histochemical assay for measurement of local glutamate concentrations in cryostat sections of rat liver. Deamination of glutamate by glutamate dehydrogenase (GDH) was coupled to the production of formazan and formazan precipitation was used for colorimetric visualization. The method was tested and validated with gelatin model sections with known glutamate concentrations. Calibration graphs showed linear relationships with high correlation coefficients (>96%) between glutamate concentrations or section thickness and absorbance values. The method was reproducible, with a constant percentage of 60 ± 5% of glutamate being converted in gelatin model sections containing glutamate concentrations of 2 mM and higher. Glutamate concentrations were estimated in periportal, intermediate, and pericentral zones of liver lobules that contain low, intermediate, and high GDH activity, respectively. In fed adult male rat livers, periportal zones contained the highest concentrations of glutamate (~14 mM) and intermediate and pericentral zones ~13 and 9 mM, respectively. On starvation, glutamate concentrations increased only in the small rim of pericentral cells that express glutamine synthetase, to ~15 mM. In livers of fetal and newborn rats, glutamate was homogeneously distributed, with a concentration of ~5 mM. In suckling rat liver, distribution of glutamate was still homogeneous but the concentration was increased to ~8 mM. These glutamate distribution patterns were in agreement with those detected immunohisto-chemically. (J Histochem Cytochem 45:1217-1229, 1997)

Key Words: glutamate, glutamate dehydrogenase, glutamine synthetase, quantitative histochemistry, image analysis, metabolic zonation, liver lobule


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