Regulation of Hepatic Metabolism: Intra- and Intercellular Compartmentation

494 Pages·1986·12.852 MB·English

by  Dieter Sasse (auth.), Ronald G. Thurman, Frederick C. Kauffman, Kurt Jungermann (eds.)

Regulation of
Hepatic Metabolism
Intra- and Intercellular
Compartment at ion
Regulation of
Hepatic Metabolism
Intra- and Intercellular
Compartmentation
Edited by
Ronald G. Thurman
University of North Carolina School of Medicine
Chapel Hill, North Carolina
Frederick C. Kauffman
University of Maryland School of Medicine
Baltimore, Maryland
and
Kurt Jungermann
Georg-August Universitiit
G6ttingen, Federal Republic of Germany
PLENUM PRESS • NEW YORK AND LONDON
Library of Congress Cataloging in Publication Data
Main entry under title:
Regulation of hepatic metabolism.
Includes bibliographies and index.
1. Liver. 2. Metabolism-Regulation. 3. Cell compartmentation. I. Thurman,
Ronald G. II. Kauffman, Frederick C. III. Jungermann, Kurt. [DNLM: 1. Cell
Compartmentation. 2. Liver-metabolism. WI 702 R344]
QPI85.R44 1986 612'.35 85-30084
ISBN-13: 978-1-4684-5043-9 e-ISBN-13: 978-1-4684-5041-5
001: 10.1007/978-1-4684-5041-5
© 1986 Plenum Press, New York
A Division of Plenum Publishing Corporation
233 Spring Street, New York, N.Y. 10013
Softcover reprint of the hardcover 1s t edition 1986
All rights reserved
No part of this book may be reproduced, stored in a retrieval system, or transmitted
in any form or by any means, electronic, mechanical, photocopying, microfilming,
recording, or otherwise, without written permission from the Publisher
Contributors
William F. Balistreri, Children's Hospital, Research Foundation, University of
Cincinnati, Cincinnati, Ohio 45229
Jeffrey Baron, The Toxicology Center, Department of Pharmacology, The Uni
versity of Iowa, College of Medicine, Iowa City, Iowa 52242
Gunnar Bengtsson, Research Laboratories of the Finnish State Alcohol Com
pany, Alko Ltd., SF-00101 Helsinki 10, Finland. Present address: National
Institute of Forensic Toxicology, N-0372 Oslo, Norway
Britton Chance, The Johnson Research Foundation, University of Pennsylvania,
Philadelphia, Pennsylvania 19104
Wolfgang Gerok, Medizinische Universitatsklinik, D-7800 Freiburg, Federal
Republic of Germany
Jorge J. Gumucio, Veterans Administration Hospital, University of Michigan,
Ann Arbor, Michigan 48109
Dieter Hiiussinger, Medizinische Universitatsklinik, D-7800 Freiburg, Federal
Republic of Germany
Sungchul Ji, Department of Pharmacology and Toxicology, College of Phar
macy, Rutgers University, Piscataway, New Jersey 08854
Kurt Jungermann, Institut fUr Biochemie, Universitat Gottingen, D-3400 Gott
ingen, Federal Republic of Germany
Norbert Katz, Institut fUr Biochemie, Universitat Gottingen, D-3400 Gottingen,
Federal Republic of Germany
Frederick C. Kauffman, Department of Pharmacology and Experimental Ther
apeutics, University of Maryland School of Medicine, Baltimore, Maryland
21202
vi Contributors
Thomas T. Kawabata, The Toxicology Center, Department of Pharmacology,
The University of Iowa College of Medicine, Iowa City, Iowa 52242
A. V. LeBouton, Department of Anatomy, College of Medicine, University of
Arizona, Tucson, Arizona 85724
John J. Lemasters, Laboratories for Cell Biology, Department of Anatomy,
School of Medicine, University of North Carolina at Chapel Hill 27514
o.
Kai Lindros, Research Laboratories of the Finnish State of Alcohol Company,
Alko Ltd., SF-OOIOI Helsinki 10, Finland
Franz M. Matschinsky, Department of Biochemistry and Biophysics, Diabetes
Research Center, University of Pennsylvania, Philadelphia, Pennsylvania 19104
Bjprn Quistorff, Department of Biochemistry, University of Copenhagen, 2200
Copenhagen, Denmark
Jan A. Redick, The Toxicology Center, Department of Pharmacology, The
University of Iowa College of Medicine, Iowa City, Iowa 52242
P.D.l. Richardson, Medical Department, Astra Pharmaceuticals Ltd., Home
Park Estate, Kings Langley, Hertfordshire WD4 8DH, United Kingdom
Mikko Salaspuro, Research Laboratories of the Finnish State School Alcohol
Company, Alko Ltd., SF-OOIOI Helsinki 10, Finland
Dieter Sasse, Institute of Anatomy, University of Basel, CH-4056 Basel,
Switzerland
Fred J. Suchy, Children's Hospital, Research Foundation, University of Cin
cinnati, Cincinnati, Ohio 45229
Ronald G. Thurman, Department of Pharmacology, School of Medicine, Uni
versity of North Carolina at Chapel Hill, North Carolina 27514
Hannu Viiiiniinen, Research Laboratories of the Finnish State Alcohol Com
pany, Alko Ltd., SF-OO 10 I Helsinki 10, Finland
Jeffrey M. Voigt, The Toxicology Center, Department of Pharmacology, The
University of Iowa College of Medicine, Iowa City, Iowa 52242
P.G. Withrington, Department of Pharmacology, The Medical College of St.
Bartholomew's Hospital, London ECIM 6BQ, England
Preface
The liver is an exceptionally complex and diverse organ that functions both as
an exocrine and an endocrine gland. It secretes bile, which contains many con
stituents in addition to bile salts, and it synthesizes and releases many substances
in response to the body's demands, including prohormones, albumin, clotting
factors, glucose, fatty acids, and various lipoproteins. It has a dual blood supply
providing a rich mixture of nutrients and other absorbed substances via the portal
vein and oxygen-rich blood via the hepatic artery. This functional heterogeneity
is accompanied by cellular heterogeneity. The liver contains many cell types
including hepatic parachymal cells, Kiipffer cells, Ito cells, and endothelial cells.
The most abundant cell type, the parenchymal cells, are biochemically and
structurally heterogeneous. The cells in the oxygen-rich areas of the portal triad
appear more dependent on oxidative metabolism, whereas those around the
central vein (pericentral, perivenous, or centrolobular areas) are more dependent
upon an anaerobic mechanism. Throughout this volume the latter three terms
are used synonymously by various authors to indicate the five to eight layers of
cells radiating from the central vein.
Structural and metabolic heterogeneity of hepatic parenchymal cells has
been demonstrated by a variety of approaches, including histochemical, ultra
structural, and ultramicrobiochemical studies. This microheterogeneity is linked
to the physiological functions of the liver and its response to injurious substances.
For example, it has long been appreciated that many drugs and chemicals damage
different regions of the liver lobule with a great deal of specificity. Mechanisms
responsible for these phenomena remain obscure. Many drugs with allyl groups
damage hepatocytes around the portal triad (periportal regions). In contrast,
acetaminophen, ethanol, and halogenated hydrocarbons damage cells near the
central vein. Mechanisms regulating the expression of different biochemical
properties of hepatocytes in different regions of the liver lobule remain poorly
defined; however, recent data indicate that zonation of metabolism is under short
and long-term control.
vii
viii Preface
The purpose of the work described in this volume is two-fold: first, to
describe the application of recently developed methods to investigate the role of
inter- and intracellular compartmentation in hepatic function and toxicity and
second, to present work that relates metabolic microheterogeneity to liver func
tion. The book is divided into five sections. The first deals with modem ana
tomical concepts of hepatic cellular heterogeneity. In addition to discussing
cellular heterogeneity, these chapters deal with current understanding of the
hemodynamics of the liver and its innervation. This discussion is followed by
several chapters which detail state-of-the-art methods that are currently being
applied in studies of hepatic biochemical micro heterogeneity. Recent applications
of these methods are highlighted. The third section of this book is comprised of
chapters by recognized authorities who evaluate metabolic zonation in liver
function and disease. A final chapter summarizes advantages and limitations of
the approaches that have been taken to investigating hepatic intracellular het
erogeneity and identifies research areas that we feel require immediate and long
term attention. The intention of this volume is to emphasize metabolic zonation
as a new dimension in hepatic function and regulation. The thorough treatment
of new methods and their application in defining metabolic microheterogeneity
of the liver is aimed at stimulating new initiatives in this exciting area of research.
Ronald G. Thurman
Frederick C. Kauffman
Kurt Jungermann
Contents
I. Liver Structure
Chapter 1
Liver Structure and Innervation
Dieter Sasse
1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
2. Liver Structure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
2.1. Development.......................................... 4
2.2. General Histology. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
2.3. Ploidy............................................... 6
2.4. Ultrastructure ......................................... 8
2.5. Sinusodal Cells. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
2.6. Architecture of Liver Parenchyma. . . . . . . . . . . . . . . . . . . . . . . . . 14
3. Innervation of the Liver . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
3.1. Macroscopic Aspects ................................... 19
3.2. Microscopic Aspects. . . .. .. ... .. . . . . . .... . .. .. . . . . . . . ... 19
3.3. Electron-Microscopic Aspects. . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
4. Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
Chapter 2
Hepatic Hemodynamics and Microcirculation
P. G. Withrington and P. D. 1. Richardson
1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
2. Hepatic Vascular Resistance: Control Values. . . . . . . . . . . . . . . . . . . . 28
Ix
x
Contents
3. Techniques. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
4. Hepatic Arterial Resistance. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
4.1. Intrinsic Regulation .................................... 30
4.2. BloodGases . . . . . . . .. . .. . . . . . . . . .. .. . . . ... . . . . . . . . . . . . 31
4.3. Osmolarity............................................ 31
4.4. Nervous Control. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
4.5. Gastrointestinal and Pancreatic Hormones .................. 32
4.6. Catecholamines and Related Drugs. . . . . . . . . . . . . . . . . . . . . . . . 33
4.7. Autacoids and Systemic Peptide Hormones ....... . . . . . . . . . . 33
4.8. Summary............................................. 34
5. Portal Venous Resistance. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
5.1. Intrinsic Regulation .................................... 35
5.2. BloodGases . . . . . . . . .. . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . 35
5.3. Osmolarity............................................ 36
5.4. Nervous Control. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
5.5. Gastrointestinal and Pancreatic Hormones .................. 36
5.6. Catecholamines and Related Drugs. . . . . . . . . . . . . . . . . . . . . . . . 37-
5.7. Autacoids and Systemic Peptide Hormones . . . . . . . . . . . . . . . . . 38
5.8. Summary............................................. 39
6. Interrelationship Between the Hepatic Artery and the Portal Vein ... 41
6.1. Mutual Pressure-Flow Curves. . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
6.2. "Transhepatic" Drug Effects ............ . . . . . . . . . . . . . . . . . 42
7. Capacitance and Exchange . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
7.1. Regulation of Liver Volume ................ . . . . . . . . . . . . . 44
7.2. Exchange in the Hepatic Microcirculation. . . . . . . . . . . . . . . . . . 47
8. Relationship of Liver Circulation to Liver Function. . . . . . . . . . . . . . . 48
8.1. Postprandial Hyperemia. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
8.2. Metabolic Stimulation .................................. 48
9. Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
II. Methods
Chapter 3
Histology and Histochemistry
Dieter Sasse
1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
2. Histology................................................. 58
2.1. Tissue Pretreatment .................................... 58
Contents xi
2.2. Routine Staining Methods. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64
2.3. Special Staining Methods. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66
3. Histochemistry............................................. 68
3.1. Substances............................................ 68
3.2. Enzymes............................................. 73
4. Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84
Chapter 4
Immunohistochemistry
Jeffrey Baron, Jeffrey M. Voigt, Thomas T. Kawahata, and Jan A. Redick
1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87
2. Tissue Preparation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89
2.1. General Considerations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89
2.2. Fixation Procedures .................................... 90
2.3. Embedding Procedures. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92
2.4. Unfixed, Frozen Sections. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93
3. Immunoenzymatic Techniques . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
3. 1. General Considerations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
3.2. Direct and Indirect Peroxidase Methods. . . . . . . . . . . . . . . . . . . . 97
3.3. Unlabeled Antibody Peroxidase-Antiperoxidase Method ...... 99
3.4. Avidin-Biotin-Peroxidase Method. . . . . . . . . . . . . . . . . . . . . . .. 101
4. Immunofluorescence Techniques. . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 102
4.1. General Considerations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 102
4.2. Direct Immunofluorescence. . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 103
4.3. Indirect Immunofluorescence. . . . . . . . . . . . . . . . . . . . . . . . . . . .. 104
5. Dual-Labeling Techniques ................................... 106
5.1. General Considerations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 106
5.2. Dual Immunoenzymatic Labeling Methods. . . . . . . . . . . . . . . . .. 106
5.3. Dual Immunofluorescence Labeling Methods. . . . . . . . . . . . . . .. 107
5.4. Combined Immunoenzymatic and Immunofluorescence
Labeling Methods. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 107
6. Quantitative Immunohistochemical Techniques. . . . . . . . . . . . . . . . . . . 108
6.1. General Considerations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 108
6.2. Microdensitometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 109
6.3. Microfluorometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 110
7. Summary................................................. 112
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 112