1Causes of Cellular Injury in Neurodegeneration
• Oxidative Stress
• Mitochondrial Dysfunction
• Excitotoxicity and calcium
• Neurotrophic factors
• Microglia and cytokines
2Necrosis
– Ion fluxes across the plasma membrane
– Activation of Ca2+-dependent proteases
– Mitochondrial disruption
– Loss of ATP
– Cellular swelling
– Preservation of nuclear DNA
3Apoptosis
– Cell shrinkage
– Maintenance of the plasma membrane
– Cytoskeletal depolymerization
– Fragmentation of nuclear DNA
– Requires protein synthesis
4Causes of cell death
– Depletion of energy stores
– Failure of ion pumps
– Neuronal depolarization
– Increases in intracellular calcium
– Free radical formation
– Acidosis
– Protein aggregation
– ER Stress Response
– Autophagy
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14Common Factors in Neurodegenerative Disease
• Oxidative stress
• Axonal Injury
• Accumulation of Cytoskeletal Proteins
• Pathology presents as a result of cell death
• Age-related onset
• Disaster theory???
15Protein Aggregation: Cell Death
• Proteasome proteolyses ubiquitinated substrates
• It is an endopeptidase complex that can be compared to a ‘trash can’
• Undegradable protein aggregates play a role in a number of neurodegenerative diseases
– Polyglutamine repeats
– Parkin
• Uncleavable aggregates lead to cellular dysfunction and death
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20Brain Anatomy and Movement Control
• Movement control operates primarily through the basal ganglia
• Anatomy and physiology of basal ganglia are extremely complicated
• Nobody really knows how it all works
• Underlying hypothesis do exist and are testable
• Much of what we do know comes from the study of disease
21Basal Ganglia Structures
• Receives input from the Cerebral cortex
• Striatum (including caudate nucleus and putamen)
• Global Pallidus (lateral and medial)
• Subthalamic nucleus
• Substantia nigra (pars compacta and pars reticulata)
• Output goes from the thalamus back to the cerebral cortex
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25Main Neurotransmitters of the Basal Ganglia
• γ-amino butyric acid (GABA)
• Acetylcholine (Ach)
• Dopamine (Da)
• Glutamate (Glu)
• Adenosine (Ad)
• Neurokinins (Substance P, SP)
• Opiod Peptides (endorphins/met-enkephalin, Enk)
26Basal Ganglia Connections
• SNC - Substantia Nigra Compacta
• STN – Subthalamic nucelus
• LGP – Lateral Global Pallidus
• SNR – Substantia Nigra Reticulata
• MGP Medial Global Pallidus

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34Direct Movement Control
• Direct movement control is the act of higher function to exact a positive change in activity (Start/Stop)
• Filled arrows are increased during movement control
• Open arrows are reduced during movement control
• Increased Glutamate outflow from Ventrolateral Thalamus leads to reinforcement of the decision to move.

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40Indirect Movement Control
• Indirect movement control is best thought of as the inhibition of non-required movement
• Thick with closed arrow (increased), Thin with open arrow (decreased)

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45Dopaminergic Transmission
• Dopamine Transporter
• Vesicular monoamine transporter
• Dopamine receptors
– Two families
– Five types
– D1 & 5 vs D2,3,4
• Monoamine oxidase (MAO)
• Catechol-O-Methyl Transferase (COMT)
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51Dopamine Receptors
• All types metabotropic receptors
• Two-main families
– D1-like - link to $G_\alpha s$- increase adenylate cyclase activity - indirect excitatory effect - D1 & D5
– D2-like - link to $G_\alpha i$ - increase phosphodiesterase activity - indirect inhibitory effect (induce IPSP) - D2, D3, & D4

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55Pharmacology of D-Receptors
• Pharmacological targeting of D-receptors forms the basis of many CNS drugs
• Anti-psychotics
– Typical (Haloperidol) and Atypical (Aripiprazole)
• Anti-emetics - metoclopramide
• Anti-arrythmics - procainamide
• Antidepressants - imipramine
• Anti-Parkinsonians - pramipexole
56Movement Disorders
• Basal ganglia
• Movement control and Neurotransmission
• Dopaminergic Signaling
– Pathway
– Receptors
– Functions
• Hypo and Hyperkinetic Diseases
– Pathology
– Dopaminergics
– Other therapies
57Hypokinesia (dyskinesia)
abnormally decreased mobility or motor function (both start and stop)
58Hyperkinesia
abnormally increased muscular activity
59Bradykinesia
slowed ability to start or continue movement
60Chorea
irregular, unpredictable, and involuntary muscle jerks (distinct form Tics which are coordinated abnormal movements)
61Athetosis
slow and writhing abnormal movements
62Dystonia
abnormal posture resulting form tonic contractions
63Parkinson’s Disease
• Classic Hypokineticdisorder
• Associated dystonia
• Essential Tremor
• Young and Older onset forms of disease
• Five stages
64Parkinson’s Disease (Causes)
• Environmental factors
– Exogenous toxins (MPTP)
– Trauma
• Genetic factors
– Low concordance rates
– Multiple potential targets
• Parkin, UCHL1, $\alpha$-synuclein, Leucine repeat rich kinase-2,
PTEN-induced kinase 1
• Mitochondrial DNA
• Italian kindred
– α-synuclein
65Genetics of Parkinson’s Disease
• Parkin - E3 ubiquitin ligase
• UCHL1 – a deubiquitinating enzyme
• $\alpha$-synuclein – SNARE chaperone interacts with tubulin synaptic rearrangement
• Tau – microtubule associated protein
• Dardarin (LRRK2) – signaling protein that operates like Ras to control autophagy and induce apoptosis
• PTEN-induced kinase 1(PINK1) –serine/threonine kinase that is thought to protect cells from mitochondrial stress
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72Dopamine Oxidation and Oxidative Stress
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78Dopamine Therapy
• Carlsson (1957) showed DA in the striatum
• Reserpine in animals induced PD by striatal loss of DA
• Ehringer & Hornykielwicz (1960) DA i.v reduced akinesis of reserpine
• Cotzias (1967) found high dose oral levodopa effective in idiopathic PD.
79Levodopa Metabolism
• Peripheral vs central conversion
• Carbidopa
– Inhibitor of AAAD
– Unable to cross Blood Brain Barrier
– Decreases peripheral conversion
– Increases central effect and reduces peripheral side effects

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81Huntington’s Disease
• Choreas have been described for over 300 years
• Formally described in 1872 by George Huntington
• Occurs in 5 to 10 per 100,000 people
• Inherited as an autosomal dominant disorder
• Caused by trinucletoide repeats in gene IT15
• Disease usually starts in mid-thirties to mid-forties
• Personality change and adventitious movement develops into chorea, cognitive dysfunction and memory loss
• Death usually occurs as a result of secondary factors within 10 to 30 years
82Excitotoxicity and Huntington’s
• Patients are often thin
– High metabolic rate??
• Striatal injection of NMDA agonists produces symptoms
• 3-nitropropionic acid (3-NP) complex II inhibitor produces striatal lesions
83Huntingtin (IT15 gene)
• Expressed in cytoplasm
• No clear function
• Binds to Huntingtin associated protein 1 (HAP1)
• Most common in NO synthesizing neurons
• Mutant form contains polyglutamine repeats (CAG)
• More repeats more severe disease
• Maybe caspase substrate
• Binds to Glyceraldehyde-3-phosphates dehydrogenase
• Polyglutamines bind to sugars causing aggregation

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87Pharmacology for Huntington's
• NMDA antagonists
• Antioxidants (Omega-3 Fatty acids)
• Minocycline (apoptosis inhibitor)
• Neurotrophic growth factors (GDNF)
• Antidopaminergic (Tetrabenazine TBZ VMAT inhibitor)
• Inhibition of aggregation
– Disaccharides bind polyQ
– Trehalose
• Antipsychotics (Fluphenazine, D2 effect)
• Rapamycin (inhibit the kinase mTOR)
88Symptomatic Pharmacology
• Early in disease depressed, irritable, anxious, or psychotic
– Fluoxetine
– Carbamazepine
– Caution can impair mobility
• Large-amplitude chorea
– Reserpine
– Benzodiazepines
• Juvenile onset and rigidity
– DA agonists
89Amyotrophic Lateral Sclerosis
• Disease onset usually occurs in mid-thirties to mid-fifties
• Presents as peripheral weakness and/or loss of function (heterogeneous presentation makes diagnosis difficulty)
• Leads to paralysis and death
• Disease progression is usually 2-3 years
• No known effective therapy
90Guamanian ALS-PDC
Guamanian ALS-PDC is a neurological disease that is characterized by a combination of symptoms of amyotrophic lateral sclerosis (ALS) and Parkinson's disease.

91Age-dependency to ALS Survival
The older you are when diagnosed, the shorter you live afterward

92Gender Bias in ALS
More men are affected by ALS than women
93SOD Mutation and ALS
It is believed that mutations in the SOD1 gene lead to the accumulation of toxic SOD1 protein aggregates in motor neurons, which ultimately results in the death of these neurons. This, in turn, leads to the progressive loss of muscle function and eventual paralysis, which are hallmark symptoms of ALS.

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100Measures of ALS Progression
• ALS-FRS
– Functional rating scale
– Considers upper and lower motor neuron loss and specific functional effects
– Disease progression related to phenotypic presentation
• Ultrasound
– Can determine loss of motor unit density
• Electromyography
– Adverse events, fasiculations, bloackage events
– Disorder recruitment
• Pre-FALS Study
– Identify biomarkers
101ALS Therapeutic Strategies
• Symptomatic
– BiPAP
– Mucolytic agents
– Antidepressants
– Mild Exercise
• Neuroprotection/Antioxidants
– Vitamin E
– Selegiline
– NAC
• Increased Mitochondrial Function
– Dexpramipexole
• Neurotrophic factors
– IGF-1
– Myotrophin
• Reduced Excitation
– Riluzole
• Increases ventilator free life
– Lamotrigine
• Voltage gated sodium channel blocker
– Ceftriaxone
• Increased glutamate uptake by astrocytic cells EAAT2 expression
– Retigabine
• Stimulate Potassium channel opening (hyperpolarize)
• Anti-inflammatory
– Mastinib
• CD117 tyrosine kinase inhibitor (myeloid cells)
102Riluzole
• Sodium channel blocker
• High voltage calcium channel blocker
• NMDA receptor antagonist
• Antagonized by pertussis toxin
– Potential GPCR
• Metabolized in liver by P450 enzymes
– Potential hepatotoxicity
• Slows down progression of disease ?
103Other Potential Therapies for ALS
• Tau-tubulin kinase inhibitors (TDP-43)
• Antisense oligos (C9orf72)
• Antisense FUS
• RAN antibodies