Table Of ContentDopaminergic Neuron Transplantation
in the Weaver Mouse Model of Parkinson's Disease
ADVANCES IN EXPERIMENTAL MEDICINE AND BIOLOGY
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NATHAN BACK, State University ofN ew York at Buffalo
IRUN R. COHEN, The Weizmann Institute ofS cience
DAVID KRITCHEVSKY, Wistar Institute
ABEL LAJTHA, N.S. Kline Institutefor Psychiatric Research
RODOLFO PAOLETTI, University ofM ilan
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DOPAMINERGIC NEURON TRANSPLANTATION IN THE WEAVER MOUSE MODEL
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Dopaminergic Neuron
Transplantation
in the Weaver Mouse Model
ofParkinson's Disease
By
Lazaros C. Triarhou
University ofM acedonia
Thessaloniki. Greece
Springer Science+Business Media, LLC
Library of Congree Cataloginh-in-Publication Data
Triarhou, Lazaros Constantinos, 1957-
Dopaminergic neuron transplantation in the weaver mouse model ofParkinson's
disease I Lazaros C. Triarhou.
p. cm. --(Advances in experimental medicine and biology; v. 517)
IncJudes bibliographical references and index.
ISBN 0-306-47418-2 (alk. paper)
1. Parkinson's disease. 2. Dopaminergic neurons--Transplantation. 3. Parkinson's
disease--Surgery. I. Title. II. Series.
RC382 .T75 2000
617.4'810592--dc21
00-046207
ISBN 978-1-4613-5201-3 ISBN 978-1-4615-0699-7 (eBook)
DOI 10.1007/978-1-4615-0699-7
©2002 Springer Science+Business Media New York
10 9 8 7 6 5 4 3 2
A c.I.P. record for this book is available from the Library ofCongress.
All rights reserved.
No part ofthis book may be reproduced, stored in a retrieval system, or transmitted
in any fonn or by any means, electronic, mechanical, photocopying, microfilming,
recording, or otherwise, without written pennission from the PubJisher.
PREFACE
Parkinson's disease, a degenerative brain disorder, affects an estimated
0.25-0.50% of the population. Symptoms result from neuronal degeneration in the
nigrostriatal dopaminergic pathway and include tremor, rigidity and gradual slow
ness of spontaneous movement. The cause of Parkinson's disease is only partially
understood; both environmental factors and a genetic predisposition have been im
plicated in its etiopathogenesis. Neural transplantation is being used experimentally
for providing an alternative biological source of dopamine both in animal studies
and in experimental clinical trials.
This book is the result of 15 years of research on the transplantation of
dopaminergic neurons in the striatum of the weaver mouse, a neurological mutant
characterized by genetically-determined degeneration ofm idbrain dopamine neurons.
The weaver mouse constitutes the only available laboratory model with a chronic
progressive disease that mimics Parkinsonism. The other two models currently used
to investigate dopaminergic mechanisms rely on the use of the neurotoxins 6-
hydroxydopamine and methylphenyltetrahydropyridine for the selective removal of
dopaminergic neurons from an otherwise healthy organism.
Structural and functional aspects of transplantation of mesencephalic dopamin
ergic grafts into the striatum of weaver mice are reviewed, including histochemical
correlates of graft survival and integration, numerical aspects of donor neuron
survival, ultrastructural findings on synaptogenesis, neurochemical indices of dopam
ine uptake function and receptor binding, gene expression of several structural and
neurotransmitter-receptor related molecules, the levels of striatal amino acid receptors,
and the behavioral effects of unilateral and bilateral neuronal transplantations.
One finding of particular interest is that weaver hosts appear to offer a subopti
mal environment for dopamine cell survival compared to wild-type hosts subjected
to 6-hydroxydopamine lesions. While the weaver brain permits the survival and
histotypic differentiation of donor dopaminergic neurons to a degree sufficient for
bringing about functional improvement, there may be micro environmental factor(s)
putatively influencing the final number of surviving dopamine cells in the grafts. It
is pertinent to mention that at birth, many young neurons undergo degeneration in
the weaver striatum just beneath the subependymal plate, and by one year of age an
estimated 22% of medium-sized striatal neurons have been lost. The possibility that
the disease process exerts an influence on graft survival has been repeatedly brought
up in clinical transplantation studies in Parkinsonism. Such considerations add value
to the weaver mutant as a model of nigrostriatal dopamine degeneration, particularly
in studying host-graft interactions.
v
ACKNOWLEDGMENTS
The author thanks all of his collaborators and co-authors of the original studies,
particularly those who have devotedly shared the laboratory bench, for making this
work possible; special thanks for that matter to Drs. Patrik Brundin, Guy Doucet,
Walter Low, Lupe Mengod, Adamantia Mitsacos, Jay Simon, Carme Sola, Lefteri
Tsoukalas and Thomas Witt. Words of deep gratitude are extended to Drs. Shirley
Bayer, Anders Bjorklund, Manuel del Cerro, Joseph Hingtgen, Elias Kouvelas and
Chema Palacios for expanding the author's scientific horizons; to Drs. Biagio
Azzarelli, Ben Boukai, Jans Muller and Sidney Ochs for inspired discussions; to the
American Society for Neural Transplantation and Repair for the exciting forum it
has created and maintained; to the Bodossakis Foundation, Athens, for honoring
this research with the bestowal of its Science Award; to the following agencies and
foundations for supporting the author's scientific efforts over the years: the
Association for the Advancement of Mental Health Research and Education, the
Ataxia-Telangiectasia Children's Project, Eli Lilly and Company, the National Ataxia
Foundation, the National Institutes of Health, and the Research and Sponsored
Programs ofIndiana University; and to Dr. Ronald Landes personally and the staff
of Landes Bioscience for their encouragement, understanding and care during the
composition of this monograph.
vii
CONTENTS
1. INTRODUCTION ............................................................................................ 1
Dopamine and Parkinson's Disease ................................................................................... 1
Experimental Models of Parkinsonism in Laboratory Animals ..................................... 5
Graft-Assisted Neural Reconstruction ("Brain ware Engineering"?) ............................. 7
References ............................................................................................................................ 8
2. BIOLOGY AND PATHOLOGY OF THE WEAVER MUTANT MOUSE 15
Introduction ....................................................................................................................... 15
Cellular and Molecular Genetics of the Weaver Mutation ............................................ 15
Alterations of the Mesotelencephalic Dopamine Projection System ............................ 16
Cerebellar Phenotype of the Weaver Mutant ................................................................. 29
Hippocampal Morphology ................................................................................................ 31
Biology of Normal H Mutant Cell Associations ............................................................. 32
Behavioral Phenotype ....................................................................................................... 34
Structural Clues to the Weaver Riddle ............................................................................ 35
References .......................................................................................................................... 36
3. HISTOCHEMICAL PROPERTIES OF INTRASTRIATAL
MESENCEPHALIC GRAFTS ........................................................... 43
Introduction ....................................................................................................................... 43
Methodological Considerations ........................................................................................ 43
Expression of Catecholaminergic Neurotransmitter-Related Molecules
and Quantitative Aspects of Dopaminergic Neuron Survival ............................... 47
Comparative Survival of Dopaminergic Neurons in Grafts Placed
in Weaver and in 6-0HDA Lesion Hosts ................................................................. 51
Expression of Neuropeptides
and Structural Proteins ............................................................................................. 53
References .......................................................................................................................... 56
4. STRUCTURAL CORRELATES OF PROCESS OUTGROWTH
AND CIRCUIT RECONSTRUCTION .............................................. 63
Axonal Reinnervation of the Host Striatum ................................................................... 63
Synaptic Investment of Graft-Derived Dopamine Terminals ........................................ 65
Compartmental Specificity of the Striatal Reinnervation ............................................. 73
Innervation of Nonstriatal Regions by the Grafts .......................................................... 74
ix
x COlltellts
Chemoaffinity and Axon-Target Recognition in Development
and in Transplantation .............................................................................................. 76
Dendrite Extension from the Graft into the Host Striatum .......................................... 77
Expression of Molecules Related to Axonal and Dendritic Outgrowth ........................ 78
References .......................................................................................................................... 83
5. NEUROCHEMICAL INDICES
OF FUNCTIONAL RESTORATION ................................................ 89
Dopamine Uptake Markers .............................................................................................. 89
Autoradiography of [3H)Dopamine Uptake .................................................................... 89
Dopamine Receptors ......................................................................................................... 96
Neurotensin Receptors ~ ................................................................................................... 100
Excitatory and Inhibitory Amino Acid Receptors ........................................................ 100
References ........................................................................................................................ 102
6. BEHAVIORAL RECOVERY
OF FUNCTIONAL RESPONSES .................................................... 107
Unilateral Grafts and Circling Behavior ....................................................................... 107
Correlation of Turning Bias with Structural and Biochemical Parameters .............. 109
Dissociation of the Functional Contribution of Graft-Derived Axons
and Dendrites in Rotational Asymmetry ............................................................... 112
Enhancement of Motor Performance
after Bilateral Transplantation ............................................................................... 118
References ........................................................................................................................ 122
7. DIRECTIONS FOR FUTURE RESEARCH ............................................. 127
Introduction ..................................................................................................................... 127
Analysis of Early Events in Graft-Host Interactions ................................................... l29
Trophic Considerations ................................................................................................... 131
Neurotransmitter Mechanisms ....................................................................................... 134
Supplemental Restoration of the Interrupted
Nigro-Striato-Nigral Loop by StriatallNigral Double Grafts .............................. 135
References ........................................................................................................................ 138
INDEX ............................................................................................................... 145
INTRODUCTION
DOPAMINE AND PARKINSON'S DISEASE
Movement control is accomplished by complex interactions among various groups
of nerve cells in the central nervous system. One such important group of neurons is
located in the substantia nigra in the ventral midbrain. Nigral neurons give rise to an
extensive network of axonal processes that innervate the basal ganglia, establishing
predominantly symmetrical synapses with dendritic spines and shafts of medium
spiny projection neurons.I,2 Neurons of the substantia nigra communicate with
neurons ofthe basal ganglia by liberating the neurotransmitter dopamine (DA). Such
an interaction at the biochemical level is responsible for the fine tuning of an
organism's movements.
Parkinson's disease or paralysis agitans3 is a neurological disorder that affects
movement control. In Parkinson's disease, neurons of the substantia nigra
progressively degenerate4 (Fig. 1); as a result, the amount of DA available for
neurotransmission in the corpus striatum is lowered.5 The biochemical imbalance
manifests with typical clinical symptoms that include resting tremor, rigidity,
bradykinesia, i.e., a gradual slowness ofs pontaneous movement, and loss ofp ostural
reflexes or, in other words, poor balance and motor coordination.6-9 An estimated
half million people are affected with Parkinson's disease and related disorders in the
United States.IO
Reductions in DA content and uptake indices have been documented in
Parkinson's disease by a variety oftechniques, including eH]mazindol bindingll or
computer-aided analyses of neuromelanin pigmentl2 in postmortem brain tissues,
as well as positron emission tomography following the administration of6-L-[18F]
fluorodopa or [iiC]nomifensine as DA uptake tracers in vivo.13 A selective increase
of N-methyl-D-aspartate (NMDA)-sensitive glutamate binding but not of (RS)-a
amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) and kainate occurs in
the striatum of postmortem brain tissue from patients with Parkinson's disease. 14
An important indirect action ofDA in the striatum may actually be the tuning
down ofthe cortical excitation of striatal neuronsY Consequently, the impairment
Department ofPathology and Laboratory Medicine, Division ofNeuropathology, Medical Science Building
A 142, Indiana University Medical Center, 635 Barnhill Drive, Indianapolis, Indiana 46202-5120.
2 L.C. TRlARHOU
Figure 1. Appearance ofnonnal (upper) and Parkinsonian (lower) human midbrain. Depigmentation of
the substantia nigra is the main macroscopic neuropathological hallmark of Parkinson's disease.
Magnification xO.7. Courtesy Dr. Jans Muller.
of dopaminergic neurotransmission that occurs in Parkinsonism may lead to an
increase in the physiological state of corticostriatal glutamatergic transmission, which
may further contribute towards reinforcing the imbalance between subsets of striatal
