A method is described for the production of detailed maps of intralobular variations of hepatocyte function and metabolite concentrations, based on variable destruction by digitonin of the lobule from the centrilobular direction. Instead of the conventional approach, in which isolated hepatocytes are then prepared and studied in suspension, perfusion is continued after digitonin treatment and the function of the unaffected lobular remnants is determined, or mean metabolite concentrations are measured by 31P-NMR. These measurements are plotted against the degree of destruction, determined precisely after each study by automated quantitative histomorphometry. These plots are transformed into curves of the function or metabolite concentration of nominal single cells at any point along the radius of the lobule. Gluconeogenesis from lactate remained stable, although reduced, even after 85–90% lobular destruction, predominated periportally and disappeared by 50% along the radius of the lobule. In 31P-NMR studies, employing 1.5 mM lactate as substrate, narrowing of the intracellular Pi resonance was observed as digitonin destruction increased; this was attributed to a decrease in the intralobular heterogeneity of the intracellular pH, which fell from approx. 7.9 to < 7.4 along the first 16% of the lobular radius (from the periportal end) and to < 7.3 in the remainder of the lobule. The ATP concentration rose, and then fell, along the radius of the lobule in a centripetal direction. The method is potentially generally applicable to a wide range of hepatocellular functions and to the measurement of metabolite concentrations, most conveniently those susceptible to estimation by NMR.
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October 1996
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Research Article|
October 15 1996
A method for determination in situ of variations within the hepatic lobule of hepatocyte function and metabolite concentrations
Shamus P BURNS;
Shamus P BURNS
*Medical Unit, St. Bartholomew's and The Royal London Hospital School of Medicine and Dentistry (Queen Mary and Westfield College, University of London), Turner Street, London E1
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Robert D. COHEN;
Robert D. COHEN
‖
*Medical Unit, St. Bartholomew's and The Royal London Hospital School of Medicine and Dentistry (Queen Mary and Westfield College, University of London), Turner Street, London E1
‖To whom correspondence should be addressed: Medical Unit, The Royal London Hospital, Whitechapel Road, London E1 1BB, U.K.
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Richard A ILES;
Richard A ILES
*Medical Unit, St. Bartholomew's and The Royal London Hospital School of Medicine and Dentistry (Queen Mary and Westfield College, University of London), Turner Street, London E1
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Jocelyn P. GERMAIN;
Jocelyn P. GERMAIN
†Department of Oral Pathology, St. Bartholomew's and The Royal London Hospital School of Medicine and Dentistry (Queen Mary and Westfield College, University of London), Turner Street, London E1
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Thomas C. H. GOING;
Thomas C. H. GOING
*Medical Unit, St. Bartholomew's and The Royal London Hospital School of Medicine and Dentistry (Queen Mary and Westfield College, University of London), Turner Street, London E1
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Stephen J. W. EVANS;
Stephen J. W. EVANS
‡Department of Epidemiology and Medical Statistics, St. Bartholomew's and The Royal London Hospital School of Medicine and Dentistry (Queen Mary and Westfield College, University of London), Turner Street, London E1
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Patrick ROYSTON
Patrick ROYSTON
§Department of Medical Statistics, The Royal Postgraduate Medical School, Hammersmith Hospital, London W12 0NN, U.K.
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Publisher: Portland Press Ltd
Received:
November 29 1995
Revision Received:
June 12 1996
Accepted:
July 02 1996
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1996
1996
Biochem J (1996) 319 (2): 377–383.
Article history
Received:
November 29 1995
Revision Received:
June 12 1996
Accepted:
July 02 1996
Connected Content
A correction has been published:
A method for determination in situ of variations within the hepatic lobule of hepatocyte function and metabolite concentrations
Citation
Shamus P BURNS, Robert D. COHEN, Richard A ILES, Jocelyn P. GERMAIN, Thomas C. H. GOING, Stephen J. W. EVANS, Patrick ROYSTON; A method for determination in situ of variations within the hepatic lobule of hepatocyte function and metabolite concentrations. Biochem J 15 October 1996; 319 (2): 377–383. doi: https://doi.org/10.1042/bj3190377
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