Review
Abstract
The causal factor(s) responsible for sporadic neurological diseases are unknown and the stages of disease progression remain undefined
and poorly understood. We have developed an animal model of amyotrophic lateral sclerosis-parkinsonism dementia complex which mimics
all the essential features of the disease with the initial neurological insult arising from neurotoxins contained in washed cycad seeds. Animals
fed washed cycad develop deficits in motor, cognitive, and sensory behaviors that correlate with the loss of neurons in specific regions of the
central nervous system. The ability to recreate the disease by exposure to cycad allows us to extend the model in multiple dimensions by
analyzing behavioral, cellular, and biochemical changes over time. In addition, the ability to induce toxin-based neurodegeneration allows us
to probe the interactions between genetic and epigenetic factors. Our results show that the impact of both genetic causal and susceptibility
factors with the cycad neurotoxins are complex. The article describes the features of the model and suggests ways that our understanding of
cycad-induced neurodegeneration can be used to decipher and identify the early events in various human neurological diseases.
q 2003 Published by Elsevier Ltd.
Keywords: Amyotrophic lateral sclerosis-Parkinsonism dementia complex; Alzheimers disease; Parkinsonism; Amyotrophic lateral sclerosis; Cycad;
Neurodegeneration; Excitotoxicity; Sterol glucoside; Animal model; Time course
Contents
1. Introduction: the fundamental problems in neurological disease research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
2. Defining neurological disease in four dimensions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
3. The Timeline concept . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
4. Overview of age-related neurological diseases. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
5. Is there a neurological Rosetta Stone?. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
6. A murine model of ALS-PDC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
7. Does the ALS-PDC model satisfy standard criteria? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
8. Genetic epigenetic interactions in the cycad model. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
9. Time line of neurodegenerative events in the cycad model. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
10. Template matching to human neurological disease . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
11. The mouse model of ALS-PDC: implications for prophylaxis and for halting disease progression . . . . . . . . . . . . . .
12. Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
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Fig. 1. Schematic: neurological disease in four dimensions. The combined X; Y; and Z-axes represent an arbitrary region of CNS affected by a progressive
neurodegenerative disorder of undefined nature. The X-axis represents cell structure/function (neuron number/structure in particular regions of CNS), Y
represents normal behavioral function, and Z represents biochemical processes in the CNS. Axis length corresponds to percent remaining function and has been
arbitrarily set for this example. As the disease progresses, the biochemical malfunction leads to greater cell loss and ultimate decreased behavioral function.
Behav: behavioral function. Cell struct: cell structure/function. Biochem: normal biochemical processes.
Table 1
Comparison of ALS, PD, AD, and ALS-PDC
ALS
PD
AD
ALS-PDC
Male:female ratio
14 [3,82]
21 [3]; 19 in Estonia [84]; ,100200 on Als and
Faroe Islands [85]; Greenland [86]; and Bulgaria [87]
39101 [89]; 401 [3]
59.3 [3]
61.9 [3]
71.9 [3]
19451960: 50 59
19951999: 65 69 [25]
ALS: amyotrophic lateral sclerosis, PD: Parkinsons disease, AD: Alzhiemers disease and ALS-PDC (ALS-parkinsonism dementia complex) are
compared.
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Fig. 2. Summary of data from the cycad mouse model. A C: behavioral deficits displayed by cycad-fed mice (A) cycad-fed mice, (B) control mice. A:
radial arm maze test of spatial memory. Reference memory (entrances into incorrect arms) and working memory (entrances to previously visited arms)
errors by cycad-fed and control mice are shown. B: Paw Print testing for walking gait length. C: Leg extension reflex test, a measure of motor neuron
integrity. D F: Cell counts or MRI volume measurements in specific CNS areas of cycad-fed mice compared to controls. D: i. Cortical thickness
measurement of several areas of cortex. V1: primary visual cortex. LEnt: lateral entorhinal cortex. M1: primary motor cortex. S1: primary
somatosensory cortex. Pir: piriform cortex. ii. Hippocampus volumes of control and cycad-fed mice. E: i. Striatum volumes of control and cycad-fed
mice. ii. Substantia nigra (SN) volumes of control and cycad-fed mice. F: i. Motor neuron counts from ventral horn of spinal cord from control and
cycad-fed mice. ii. Ventral horn volumes of control and cycad-fed mice. GI: Histological display of cell pathology in CNS of cycad-fed mice. G:
TUNEL labeling in cycad-fed mouse. i, ii: TUNEL labeling in cortex. iii. TUNEL labeling in Dentate gyrus. iiii 40 magnification. H: Caspase-3
labeling of cycad-fed mouse substantia nigra (SN). i. 10 magnification. ii. 40 magnification. I: Cresyl Violet staining of spinal cord motor
neurons. i. Control, ii, iii. Cycad-fed mouse. JL: Examples of selected biochemical changes in CNS tissue from cycad-fed and control mice. J:
Immunolabeling of GLT-1 in primary motor cortex,. i: Control, ii. Cycad-fed. K: Tyrosine hydroxalase (TH) labeling of striatum. i: Control, ii. Cycadfed. L: Immunolabeling of GLT-1 of spinal cord. i: Control, ii. Cycad-fed. J, L: scale bar 80 mm.All graphs show means ^ SEM, (* P , 0:05: A C,
G I: Original data from [7]; D F: Original data from [81]; J, L: Original data from Ref. [44]; K: Original data from Ref. [45]).
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(Fig. 2A) tasks were degraded, with corresponding neurodegeneration seen in regions of cerebral cortex and
hippocampus (Fig. 2D). In addition, the olfactory system
showed a significant loss of function accompanied by
disrupted structures and cell loss in the olfactory glomeruli.
In various regions, key molecules associated with
neuronal degeneration were altered. These included an
elevation of tau protein and various protein kinases, notably
various PKC subtypes and CDK5. In addition, elements of
the glutamatergic system were severely affected, most
notably a dramatic decrease in levels of two variants of the
GLT 1 glutamate transporter (EAAT2) (Fig. 2J and L)
accompanied by a decrease in NMDA and AMPA receptor
binding [44]. The apparent down-regulation in GLT and
glutamate receptor subtypes was noted in regions of the
CNS showing neural degeneration. Labeling for tyrosine
hydroxylase revealed a significant decrease of labeled
terminals in striatum and of cell bodies in substantia nigra
pars compacta of cycad fed mice (Fig. 2K) [45].
All of the features described above are consistent with
features of ALS-PDC, as well as key aspects of AD, PD, and
ALS. Of particular significance is the observation that the
alterations in behavior and CNS morphology are progressive in adult mice even after cycad exposure ends [7].
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Table 2
Disease progression of human and animal modeled neurodegenerative disorders
Human (post-diagnosis
progression)
AD
PD
ALS
Exponential increase of
Amyloid B deposits in
PS1 APP transgenic AD
mouse [67]
Dopaminergic degeneration in PD
appears to slow down during course
of the disease [64]
Exponentially decreasing risk in
chemically induced model of PD
(Fig. 3D) [63]
Disease progression rates are compared for Alzheimers disease (AD), Parkinsons disease (PD) and amyotrophic lateral sclerosis (ALS). Human disease rates
are determined from beginning of behavioral symptoms, while animal models measure progression from onset to death.
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starts from the initial insult and moves forward in time to the
end state condition, while the latter attempts to reverse
engineer the earliest stages from the end state. Neither data
set alone is sufficient, but the hope is that the future success
of our model will allow us to fill in the enormous gaps that
now exist concerning the timeline of pathological events in
human neurological diseases.
As discussed above, the problem is that we have no
timeline to speak of for tracing the course of age-dependent
human neurological diseases. How, for example, can we
know who is pre-symptomatic for any of these diseases,
especially any of the sporadic varieties? In principle, various
brain scans or biomarkers could be widely employed, but the
first is not economically feasible for the population at large;
the second would depend on the identification of crucial
molecules, most of these still unknown. The necessary
crossover and template matching from an animal model to
human neurological disease states is a twofold process. First,
we have to be able to place what we know from our model
system into a clear description of events in all four
dimensions. Second, armed with this information, we can
then attempt template matching to pre- and post-clinical
human patients. This procedure may be a time consuming
one, yet offers, in our view, the only realistic possibility for
prevention and early treatment of human neurological
disease. We discuss a potential strategy in more detail below.
11. The mouse model of ALS-PDC: implications for
prophylaxis and for halting disease progression
Our work has demonstrated the following: First, we have
been able to show that a toxin contained in cycad is able to
kill neurons. Mice fed with washed cycad develop the same
behavioral and pathological outcomes as ALS-PDC victims.
Second, we have been able to show clear interactions of
cycad toxicity with various susceptibility (APO E) or causal
(mSOD) gene abnormalities. Third, we have begun the long
process of defining the various stages leading from toxic
insult to neurodegeneration.
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12. Conclusion
ALS-PDC is a unique disease that displays aspects of the
major progressive neurodegenerative diseases, AD, PD and
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