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Anxiety Disorders

Pharmacotherapy
MAOI
TCA
SSRI
Lithium
CRF, SP antagonists
Benzodiazepines
TCA
SSRI
CRF, SP antagonist

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Genetic and Environmental Factors in Anxiety Disorders

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"Behavioral validation"
Behavioral validation
Multiple behavioral and physiological tests
Factor analysis (covariance analysis)
Pharmacological validation
Response to therapeutically active drugs but not to inactive drugs
Comparable pharmacokinetic profiles

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Anatomy of the serotonergic system

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Construction of 5-HT1AR KO mice

5-HT1A KNOCKOUT

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Does forebrain receptor reverse the phenotype of the 5-HT1A KO mice?

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Receptor OFF in adulthood

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Receptor ON at P21

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Forebrain Rescue
Forebrain receptor sufficient to reverse knockout phenotype
Adult receptor not required to maintain rescue
Expression during development critical to rescue

Evidence for developmental role of 5-HT

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Increased excitability in CA1 pyramidal neurons of knockout mice

5-HT1AR function:
 development vs. adulthood

long term molecular and morphological changes

Mechanism of action of antidepressants
Change of the setpoint of monoamine transmission
Plastic changes occurring in the limbic target areas of monoaminergic projections (Hipp., Amy., Ctx)
Halt hippocampal atrophy
Prevent stress-induced dendritic shrinkage
Increase hippocampal neurogenesis

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Hippocampal Atrophy In Recurrent Major Depression

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Behavioral tests to detect antidepressant action
Tail suspension test
Forced swimming test
Ultrasonic vocalization
Learned helplessness
Chronic unpredictable stress
Novelty-suppressed feeding

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Parkinson’s Disease: Clinical Characteristics
Disease of aging:
-Incidence: Overall  20/100,000
Age 70  120/100,000
Clinical Features:  tremor, rigidity, slowness of movement, postural instability
Pathology:  substantia nigra pars compacta, locus coeruleus, nucleus basalis of Meynert, olfactory bulb

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Parkinson’s Disease: Environment
1918: influenza pandemic
1983: MPTP

MPTP Induced Parkinsonism
Clinical: All cardinal manifestations of idiopathic PD.
Pharmacology: Respond to dopaminergic drugs
Pathology: Ventral substantia nigra, locus coeruleus, hypothalamus
Pathogenesis: Mitochondrial dysfunction, oxidative stress

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Parkinson’s Disease: Genetics

Parkinson’s Disease and a-Synuclein
Mutations produce autosomal dominant PD
Abundant in Lewy bodies
Present in Glial Cytoplasmic Inclusions (MSA)
Accumulates in Hallevordan-Spatz disease

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a-Synuclein Biology
Highly abundant in CNS
Concentrated in presynaptic terminals near vesicles
Lipid and vesicle binding
Conformational change upon lipid binding
Modulation of rate of recycling of the readily releasable pool

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a-Synuclein Function: Vesicles

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What is the nature of a-synuclein dysfunction that leads to dopamine neuron degeneration ?

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a-Synuclein: Gene Targeting Strategy

a-Synuclein:
Knock Out Characterization

MPTP Regimens

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a-Synuclein Null Mice:
Resistance to MPTP-Induced Neurodegeneration

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An Inducible Model of HD

A brief introduction to HD (1):
George Huntington, 1872
Age of onset: 40 to 50 yrs
Progressive
Motor: chorea, dystonia
Psychiatric: depression, anxiety, suicide
Cognitive changes: declarative memory, dementia
Autosomal dominant inheritance

Striatal specific atrophy of HD

A brief introduction to HD (2):
The HD gene, IT15 (HDCRG, 1993)
Promoter: house keeping gene?
Conserved to Fugu puffer fish
Gene KO studies
Nasir et al., Zeitlin et al., Duyao et al.
Gene product, huntingtin (htt)
Large protein in complex
Function: unknown
Ubiqutious expression
Cleavage by calpain and caspase-3

A brief introduction to HD (3):
Triplet repeat expansion (C-A-G)
Unaffected, n= 4 to 35; Pathogenic, n > 37
Triplet repeat disorders (17)

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Effect of CAG expansion on htt
CAG expansion is translated
CAG encodes for glutamine (Q)
No loss of function
Patients homozygote for HD mutation
Dominant gain of function

Mouse models of HD:
Bates’ R6 transgenic (R6/2)
Mangiarini et al. (1995) Cell
cDNA transgenic
Reddy et al. (1998) Nat Genet 20, 198-202
N171 transgenic
Schilling et al. (1999) Hum Mol Genet 8, 397-407
YAC transgenic
Hodgson et al. (1999) Cell 23, 181-92
(CAG)n knock-in
Wheeler et al. (1999) Hum Mol Genet 8, 115-22
Shelbourne et al. (1999) Hum Mol Genet 8, 763-74
Lin et al. (2001) Hum Mol Genet 10, 137-44

"Davies et al., 1997"
Davies et al., 1997
Intranuclear aggregates
“Dark bodies” (Roizin, 1977)
DiFiglia et al., 1997
Intranuclear aggregates
cytoplasmic aggregates
Scherzinger et al., 1997
Ribbon-like morphology

Question:
 Is continuous expression of the transgene required for the HD-like phenotype?
Is the HD-like phenotype reversible?

Inducible mouse model
A model with a switch

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Intranuclear & cytoplasic aggregates

Neuropathology in HD94 mice

HD mice demonstrate a progressive clasping phenotype

Summary
Expression of exon1 CAG94 leads to an HD-like phenotype

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Protocol
2 mg dox/ ml 5% sucrose
Duration: 16 weeks
Halt progression
Amelioration

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Summary
Expression of exon1 CAG94 leads to an HD-like phenotype
Abolishing gene expression for 16 weeks leads to a reversal of aggregate formation and motor phenotype

Primary striatal cultures from HD94 mice
Kinetics of aggregate formation and reversion
Possible mechanisms for aggregate reversion

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Summary (2)
In vitro
“Soluble” exon1CAG94 clears within 2 to 3 days of shutting down gene expression.
Aggregates clear within 5 days of shutting down gene expression (3d + 2d).
Aggregate clearance is inhibited in the presence of lactacystin (proteasome).

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