Abstract
It is well established that parenchymal cells in different zones of the hepatic lobule differ in function, biochemical properties, and susceptibility to hepato-toxins.1–3 The experimental approaches that have been used most extensively over the years to describe the biochemical heterogeneity of the liver lobule are classic microscopic histochemistry and quantitative histochemistry. The object of this chapter is to describe the latter approach and illustrate its application to analyses of selected substrates and enzyme activites in microdissected samples from specific zones of the liver lobule. Methods of high specificity and sensitivity are required for such analyses because the number of cells that can be obtained conveniently from specific zones of the liver lobule range in number from about 100 to 500 depending on the age and species of animal studied. The average mass of a rat liver cell is estimated to be on the order of 1 × 10-9 g. Thus, an intermediate such as ATP, which is estimated to be about 5 mmoles/kg wet weight liver, would be about 5 × 10-15 mole/cell or about 0.5–2.5 × 10-12 mole/microdissected sample. Although this amount of substrate is below the range detected by conventional biochemical analytical techniques, this level of substrate can be measured conveniently via pyridine-nucleotide-dependent enzymatic cycling assays4,5 and some bioluminescence assays.6,7 Each of these types of analyses can be applied to specimens obtained by quantitative histo-chemical sampling procedures.
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© 1986 Plenum Press, New York
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Kauffman, F.C., Matschinsky, F.M. (1986). Quantitative Histochemical Measurements within Sublobular Zones of the Liver Lobule. In: Thurman, R.G., Kauffman, F.C., Jungermann, K. (eds) Regulation of Hepatic Metabolism. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-5041-5_5
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DOI: https://doi.org/10.1007/978-1-4684-5041-5_5
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