Aging Methods and Protocols

377 Pages·2000·1.693 MB·English

by  Bernard L. Strehler (auth.), Yvonne A. Barnett, Christopher R. Barnett (eds.)

Aging Methods and Protocols
M E T H O D S I N M O L E C U L A R M E D I C I N ETM
John M. Walker, SERIES EDITOR
52. Atherosclerosis Methods and 39. Ovarian Cancer: Methods and
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M E T H O D S I N M O L E C U L A R M E D I C I N ETM
Aging Methods and
Protocols
Edited by
Yvonne A. Barnett
Christopher R. Barnett
School of Biomedical Sciences, University of Ulster
Northern Ireland
Humana Press Totowa, New Jersey
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Main entry under title:
Methods in molecular medicine™.
Aging methods and protocols / edited by Yvonne A. Barnett, Christopher R. Barnett
p. cm. —(Methods in molecular medicine ; 38)
Includes bibliogrpahical references and index.
ISBN 0-89603-582-4
1. Aging—Molecular aspects—Laboratory manuals. I. Barnett, Yvonne A. II. Barnett,
Christopher R. III. Series.
QP86.A375 1999
571.8'78–dc21 99-23842
CIP
Preface
Aging is an almost universal process within biological systems, one which
leads to a decline in functional capacity, disease onset, and eventually death.
There has been much interest in recent years to elucidate the molecular mecha-
nisms that underlie the aging process. Many theories have been proposed since
the last century that aim to explain the causes of aging. There is no one theory
that completely satisfies the phenotype of aging, but genetics and environmen-
tal factors play an important role in the etiology of age-related pathologies and
the aging process. However, there is still much to be learned about the aging
process which has been termed one of the last great frontiers in biology. Demo-
graphic changes worldwide are leading to increased average life expectancies
within our populations. These changes in population characteristics will impact
upon the economies of the supporting society, with increasing healthcare and
infrastructural costs arising from the prevalence of age-related pathologies and
other physical disabilities associated with advancing years. Many researchers
worldwide are working in the attempt to identify key cellular processes through
which it might one day be possible to slow down the aging process and thus
increase the health span of humans.
Numerous research projects—from the cellular through to tissue, organ,
and whole organism studies—are currently underway to investigate the multi-
factorial aging process. In Aging Methods and Protocols we present a number
of protocols, described by recognized experts in their field, which are being/
have been used to further our understanding of aging processes. The aim of the
editors here has been to provide a flavor of recent research advances in as many
areas of biological gerontology as space permitted in this single volume. The
editors regret that exigent circumstances may have caused under- or even
nonrepresentation of a number of equally exciting aging research areas.
The editors were delighted that Professor BL Strehler, who needs no introduc-
tion to those familiar with the field of biological gerontology, agreed to contribute
an introductory chapter to this text entitled Understanding Aging. Over several
decades, he has himself made, and continues to make, a significant contribution to
our understanding of the molecular, chemical, and biological processes that contrib-
ute to aging. The editors encourage all readers to obtain a copy of the long-awaited
third edition of Professor Strehler’s book Time,Cells and Aging, for further in-
v
vi Preface
sights into the landmark findings within aging research. A big thank you for
your inspiration and support Bernie!
The remainder of the volume is dedicated to the presentation of detailed
protocols that have been/are being used in aging research. We begin by present-
ing protocols for the study of factors associated with cell senescence and cell
death.
It has been proposed that alterations in the body’s ability to metabolize
xenobiotics and/or to defend itself or respond to biomolecule damage plays a
critical role in the accumulation of damaged biomolecules in vivo as a function
of age. A number of protocols that explore these aspects are presented. In
addition, we have included a section on the identification and quantification of
biomolecular damage (nucleic acid, protein, and lipid modifications are described)
in vivo.
The idea that mutations within mitochondrial DNA might lead to mito-
chondrial dysfunction, and thus to cellular decline/death has received much
attention in recent years. We have included a number of protocols designed to
measure the morphological, functional, and characterization of the molecular
changes that accumulate as a function of age within mitochondria.
The process of immunosenescence (age-related decline in the immune sys-
tem) has been proposed to account for the increasing incidence of morbidity and
mortality associated with infection, cancer, and possibly autoimmune phenomena
with increasing age in humans. This volume includes a description of protocols
that enable genetic and functional characteristics of immune cells and the immune
system to be determined.
The final section of Aging Methods and Protocolspresents two case stud-
ies. The first is on the role of dietary restriction in life span extension, to date
the only recognized way through which life span (health span) can be extended
in rodents and primates. The second is how we can establish and utilize trans-
genic animals for the elucidation of the molecular aspects of aging.
The editors wish to thank each of the contributors to this volume and hope
that the readership find our work to be stimulating and informative for their own
research interests.
Yvonne A. Barnett
Christopher R. Barnett
Contents
Preface .............................................................................................................v
Contributors....................................................................................................vii
1 Understanding Aging
Bernard L. Strehler...............................................................................1
PART I: THE STUDY OF CELL SENESCENCE AND CELL DEATH
2 Use of the Fibroblast Model in the Study of Cellular Senescence
Vincent J. Cristofalo, Craig Volker, and Robert G. Allen...............23
3 Human T-Cell Clones
Graham Pawelec..................................................................................53
4 Telomeres and Replicative Senescence
Hector F. Valenzuela and Rita B. Effros...........................................63
5 Detection of Molecular Events During Apoptosis by Flow Cytometry
Ruaidhri J. Carmody, Ana P. Costa-Pereira, Sharon L. McKenna,
and Tom G. Cotter...........................................................................71
6 Raf-1 Protein Kinase Activity in T Cells from Aged Mice
Christopher J. Kirk and Richard A. Miller........................................85
7 Identification of Differentially Expressed Genes in Young
and Senescent T Cells
Andrea Engel, Mahdi Adibzadeh, and Graham Pawelec................97
PART II: AGE-RELATED CHANGES TO XENOBIOTIC METABOLIZING
ENZYME SYSTEMS
8 Xenobiotic-Metabolizing Enzyme Systems and Aging
Christopher R. Barnett and Costas Ioannides...............................119
PART III: AGE-RELATED CHANGES TO DEFENSE SYSTEMS AGAINST
BIOMOLECULE DAMAGE
9 Assessing Age-Related Changes in Antioxidant Status: The FRASC
Assay for Total Antioxidant Power and Ascorbic Acid Concentration
in Biological Fluids
Iris F. F. Benzie and John J. Strain.................................................133
vii
viii Contents
10 Measurement of DNA Damage and Repair Capacity as a Function of
Age Using the Comet Assay
Peter H. Clingen, Jillian E. Lowe, and Michael H. L. Green.........143
11 Measurement of DNA Damage and Repair in Human
White Blood Cells by an Immunochemical Assay
Govert P. van der Schans.................................................................159
PART IV: AGE-RELATED CHANGES TO CELLULAR BIOMOLECULES
12 Measurement of 8-Oxo-deoxyguanosine in Lymphocytes,
Cultured Cells, and Tissue Samples by HPLC with
Electrochemical Detection
Sharon G. Wood, Catherine M. Gedik, Nicholas J. Vaughan,
and Andrew R. Collins..................................................................171
13 Mutation and the Aging Process: Mutant Frequency at the HPRT
Gene Locus as a Function of Age in Humans
Yvonne A. Barnett and Christopher R. Barnett.............................179
14 Somatic Mutations and Aging: Methods for Molecular Analysis
ofHPRTMutations
Sai-Mei Hou........................................................................................189
15 Assessment of Susceptibility of Low-Density Lipoprotein
to Oxidation
Jane McEneny and Ian S. Young.....................................................199
16 Measurement of Pentosidine in Biological Samples
Jesus R. Requena, David L. Price, Suzanne R. Thorpe,
and John W. Baynes.....................................................................209
PART V: MITOCHONDRIAL MUTATION AND FUNCTION WITH AGE
17 Causes and Consequences of Damage to Mitochondria:
Morphological Aspects
Jaime Miquel and Carlo Bertoni-Freddari......................................221
18 Causes and Consequences of Damage to Mitochondria:
Study of Functional Aspects by Flow Cytometry
Federico V. Pallardo, Juan Sastre, Jaime Miquel,
and José Viña................................................................................237
19 Analysis of Mitochondrial DNA Mutations: Deletions
Robert W. Taylor, Theresa M. Wardell, Emma L. Blakely,
Gillian M. Borthwick, Elizabeth J. Brierley,
and Douglass M. Turnbull............................................................245
Contents ix
20 Analysis of Mitochondrial DNA Mutations: Point Mutations
Robert W. Taylor, Richard M. Andrews, Patrick F. Chinnery,
and Douglass M. Turnbull............................................................265
PART VI: IMMUNE SYSTEM AND AGING
21 Assessment of T-Cell Function in the Aged: T-Cell Proliferative
and T-Cell Adherence Assays
Ian Beckman.......................................................................................281
22 Dendritic Cells in Old Age
Beatrix Grubeck-Loebenstein, Maria Saurwein-Teissl,
and Nikolaus Romani....................................................................291
23 Age-Related Alterations to Natural Killer Cell Function
Erminia Mariani, Corona Alonso, and Rafael Solana....................311
24 Immunogenetics and Lifespan: HLA
Derek Middleton, Martin D. Curran, and Fionnuala Williams.......321
PART VII: CASE STUDIES
25 Dietary Restriction and Life-Span Extension
Byung Pal Yu......................................................................................353
26 The Use of Genetically Engineered Mice in Aging Research
Julie K. Andersen..............................................................................361
Index .........................................................................................................379
ix
Contributors
MAHDI ADIBZADEH • Tuebingen Ageing and Tumour Immunology Group,
Section for Transplantation Immunology and Immunohaematology,
Department of Haematology, Oncology and Immunology, Tuebingen
University Medical School, Tuebingen, Federal Republic of Germany
ROBERT G. ALLEN • Center for Gerontological Research, Allegheny University
of the Health Sciences, Philadelphia, PA
CORONA ALONSO • Department of Immunology, Faculty of Medicine, Renia
Sofia University Hospital, University of Córdoba, Córdoba, Spain
JULIE K. ANDERSEN • Andrus Gerontology Center, University of Southern
California, Los Angeles, CA
RICHARD M. ANDREWS • Departments of Neurology and Opthalmology, The
Medical School, University of Newcastle upon Tyne, Newcastle upon Tyne,
UK
CHRISTOPHER R. BARNETT • School of Biomedical Sciences, University of
Ulster, Cromore, Road, Coleraine, County Londonderry, Northern Ireland
YVONNE A. BARNETT• School of Biomedical Sciences, University of Ulster,
Cromore, Road, Coleraine, County Londonderry, Northern Ireland
JOHN W. BAYNES • Departments of Chemistry and Biochemistry and
Opthalmology, University of Southern Carolina, Columbia, SC
IAN BECKMAN • Department of Immunology, Fremantle Hospital, Fremantle,
Perth, Western Australia
IRIS F. F. BENZIE • Department of Nursing and Health Sciences, The Hong
Kong Polytechnic University, Kowloon, Hong Kong SAR, China
CARLO BERTONI-FREDDARI • Neurobiology of Aging Laboratory, INCRA, Ancona,
Italy
EMMA L. BLAKELY • Department of Neurology, The Medical School, University
of Newcastle upon Tyne, Newcastle upon Tyne, UK
GILLIAN M. BORTHWICK • Department of Neurology, The Medical School,
University of Newcastle upon Tyne, Newcastle upon Tyne, UK
ELIZABETH J. BRIERLEY • Departments of Neurology and Geriatric Medicine, The
Medical School, University of Newcastle upon Tyne, Newcastle upon Tyne, UK
RUAIDHRI J. CARMODY • Tumour Biology Laboratory, Department of Bio-
chemistry, University College, Cork, Ireland
PATRICK F. CHINNERY • Department of Neurology, The Medical School,
University of Newcastle upon Tyne, Newcastle upon Tyne, UK