Introductory course in Neuroscience

Neuropharmacology

2. April 2012
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Prof. Jean-Marc Fritschy

A paradigm shift: role of dopamine in behavior

Effect of L-DOPA in dopamine-depleted rabbits

A. Carlsson (1960)

Emotional responses activate the same brain regions as actual sensory stimuli

Contents

Basic principles of neuropharmacology and classification of brain diseases Monamines: properties and relevance for neuro- and psychopharmacology Antipsychotic drugs Experimental approaches to study brain diseases:
Molecular basis of the sedative and anxiolytic action of benzodiazepines

Basic principles of neuropharmacology

CNS diseases affect a large fraction of the general population and have a very high social cost The pathophysiological mechanisms underlying most brain disorders are poorly understood Many CNS disorders have a genetic basis. The elucidation of mutations in familial forms of these diseases contributes markedly to our understanding of their pathophysiology Pharmacological treatments are mainly symptomatic and the mechanism of drug action is often unknown Neuropsychiatric disorders are difficult to reproduce in animal models
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Experimental approaches to study brain diseases

Genetic screening of familial forms of disease to identify genes contributing to the pathophysiology Large scale genetic screening for identifying diseasesusceptibility genes Molecular and cell biological studies in vitro to understand the function of implicated genes In vivo animal models (lesion, pharmacological treatment, targeted gene mutations)

Classification of brain diseases (1) 1. Psychiatric diseases

Neurodevelopmental disorders (autism, Rett syndrome, X-linked mental retardation, attentiondeficit disorders) Anxiety (panic, generalized anxiety, phobia, posttraumatic stress disorder) Mood disorders (depression, bipolar disorder) Schizophrenia, Tourettes disease Drug dependence

Classification of brain diseases (2) 2. Neurological diseases

Stroke and ischemia Brain lesions (trauma, tumors, infections) Epilepsy Chronic pain Sleep disorders Movement disorders (dystonia; tremor)

3. Autoimmune diseases
Multiple Sclerosis (MS) Myasthenia gravis

Classification of brain diseases (3) 4. Neurodegenerative diseases

Alzheimer Parkinson Huntington Fronto-temporal lobe dementia (FTLD) Amyotrophic lateral sclerosis (ALS) Prion diseases (Creutzfeld-Jacob)
The cause of neurodegeneration is not established but is often linked to the production of protein aggregates (e.g. -amyloids), due to abnormal proteolytic processing or to mutations (e.g., trinucleotide repeats)

Examples of diseases caused by trinucleotide repeats

Disease Fragile X syndrome Myotonic dystrophy Spinobulbar muscle dystrophy Huntington Repeat (CGG)n in FMR1 gene (CTG)n in myotonin-protein kinase gene (CAG)n in androgen-receptor gene (CAG)n in Huntingtin gene

Huntingtons disease
Autosomal dominant disorder causes by mutation in the huntingtin gene (short arm of chromosome 4) Onset in middle adulthood (minor motor coordination problems, involuntary jerking progressing towards major deterioration) Cognitive alterations and changes in personality (impulsivity, depression, psychotic symptoms) Molecular basis: presence of tri-nucleotide repeats (37 86) coding for glutamine (CAG). Longer repeats lead to early onset and more severe symptoms Physiological role of huntingtin is unknown
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Pathophysiology of Huntingtons disease

Progredient loss of GABAergic neurons in putamen and caudate nucleus Upregulation of GABAA receptors in target regions (globus pallidus) Selectivity of degeneration is not explained
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Lessons from Huntingtons disease

Power of reverse genetics: the affected gene could be identified without knowledge about ist function The pathophysiology of the disease is not due to an obvious dysfunction in the striatum The selectivity of neurodegeneration remains unexplained The path to therapy is very long and no goal is in sight In the case of complex genetic diseases, such as psychiatric diseases, the development of novel therapies based on genetic information might be even more difficult

Monoamines: relevance to psycho- and neuropharmacology

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Monoamines

Catecholamines (dopamine, noradrenaline, and adrenaline) are derived from tyrosine Serotonin (an indolamine) is derived from tryptophan They are the neurotransmitter of small groups of neurons in the brainstem that innervate most of the brain Monoaminergic neurons regulate brain state and function (neuroendocrine systems, sleep-wake cycle, motor functions, sensory perception, emotions, attention, memory)

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Catecholamine synthesis

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Common principles of monoaminergic transmission

Local synthesis and storage in vesicles Ca++-dependent release Termination of synaptic transmission by re-uptake. The transporter proteins are a major drug target Action on a multitude of receptors (mainly 7 TM domain receptors coupled to G-proteins) Presence of pre- and postsynaptic receptors Complex metabolism, in neurons, glial cells, and other tissues (Monoamine oxidase A and B); metabolites can be toxic

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Significance of monoaminergic transmission in neuro- and psychopharmacology

Monoaminergic transmission is the target of many psychoactive drugs (antidepressants, antipsychotic drugs, some psychostimulant and psychotropic drugs, and anti-parkinson drugs) Many unwanted side effects of psychopharmacological treatment arise from interactions with monoaminergic transmission Dysfunction of dopaminergic systems underlie multiple neurological and psychiatric disorders, as well as drug addiction

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Complex pharmacology of monoaminergic systems

Direct drug effects by receptor activation/inhibition (agonist, antagonist) Indirect effects by enhancing effects of endogenous transmitter (increased release (e.g., amphetamin, inhibition of re-uptake (e.g. cocain), inhibition of catabolism) Complex interactions with precursor (e.g., L-Dopa) or pseudo-transmitters (e.g., -Methyl-Tyrosin) Inhibition of catabolism can affect pseudo-transmitters present in food Complex regulation of receptors (super-sensitivity, desensitization, opposite action of pre- and postsynaptic receptors Importance of target selectivity
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A dopaminergic synapse

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Dopamine receptors

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Dopamine

Major dopaminergic projections

Mesostriatal projection Mesolimbic projection

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cc Ca

Thalamus

ic Pu GPe GPi

Insula

STN Hypoth SNc Hip SNr

DA receptor agonist-induced rotation in lesioned animals

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Opposite rotation effects caused by amphetamine

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Serotonin (5-Hydroxytryptamine)

Source:
Enterochromaffin cells, thrombocytes, neurotransmitter in central neurons (raphe nuclei)

Effects
Periphery: complex actions on the cardiovascular system, increased motility of the gastrointestinal tract, vasoconstriction CNS: Regulation of blood pressure, temperature, appetite, sleep-weak cycle, motor activity, pain perception, emotional behavior (serotonin reuptake inhibitors are antidepressants)
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Serotonergic neurons

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Functional and pharmacological relevance of the serotonergic system

Serotonin exerts multiple, complex actions by activating pre- and postsynaptic receptors coupled to various signal transduction pathways Serotonin regulates mood, attention, sleep-wake cycle, descending pain control, motor systems, autonomic functions, neuroendocrine systems Selective 5-HT reuptake inhibitors (SSRI) are widely used antidepressants Psychostimulants, recreational drugs, and hallucinogens have mixed actions of the serotonergic, noradrenergic, and dopaminergic system.
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Noradrenergic neurons: Locus coeruleus

Immunhistochemistry of dopamine--hydroxylase

Nissl staining

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Noradrenergic projections from the locus coeruleus

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Scheme of serotonergic and adrenergic synapses

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Pathogenesis of Parkinsons disease

Advancing degeneration of nigrostriatal dopaminergic neurons Formation of Loewy bodies (intracellular aggregates) in catecholaminergic neurons Idiopathic form: unknown etiology Symptomatic forms:
Side-effects of chronic neuroleptic treatment Poisoning (CO, methanol, MPTP, rotenone)

Pharmacotherapy of Parkinsons disease

Dopamine receptor agonists (D1 and D2) Apomorphin, Bromocriptin, Cabergolin, Lisurid, Dihydroergocriptin, Pergolid, Ropinirol, etc. L-Dopa Mode of action: L-Dopa is transported across the blood-brain barrier and converted into dopamine (mainly in dopaminergic neurons) Is given in combination with inhibitors of Dopadecarboxylase (Carbidopa, Benserazid) to minimize peripheral side effects

Metabolism of L-Dopa

Antipsychotic drugs Treatment of schizophrenia Typical (Dopamine D2 receptor antagonists)

Phenothiazine derivatives (Chlorpromazine) Butyrophenone (Haloperidol) Numerous side-effects (blockade of Ach, NA, 5-HT receptors): sedation, autonomic dysfunction, involuntary movements

Atypical (mechanism of action unknown)

Clozapine Risperidone
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Effects of antipsychotics on the DA system

Location Mesolimbic, mesocortical pathway Nigrostriatal pathway Tuberoinfundibular projection Medulla oblongata (area postrema)

Clinical effects (due to blockade of D2 receptors) Antipsychotic action by modulation of neuronal circuits and regulation of excitatory-inhibitory balance (only positive symptoms) Extrapyramidal motor symptoms (EPS): Parkinson-like symptoms and dyskinesia Gynaecomasty, milk secretion (due to increased prolactin secretion) Anti-emetic effects

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Extrapyramidal motor symptoms

Syndrom Early dyskinesia Symptoms Head and neck muscle spasms Prevalence, duration 5%, at start of therapy 20-30% up to 8 weeks 25% up to12 weeks 20% After months/years rare before 2 weeks Treatment Anti-cholinergic drugs Dose, anti-cholinergic drugs Dose benzodiazepines Change to clozapin; no anti-cholinergic drugs Stop therapy; dentrolene

Parkinsonoid

Akinesis, rigor, tremor, hypersalivation Agitation, restlessness Chronic hyperkinetic syndrome (irreversible); stereotypic movements of the lips, tongue, jaws Acute emergency (rigor, akinesis, high fever, tachycardia, coma)
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Akathisia Late dyskinesia

Malignant neuroleptic syndrom

Relationship between wanted and unwanted effects of neuroleptics

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Tests for cognitive function

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Predicting schizophrenia: a longitudinal study

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Differences in gray matter volume prior to first psychosis

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Differences in gray matter volume after onset of psychosis

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New hypotheses about the pathophysiology of schizophrenia

Structural anomalies in the brain can be shown prior to the first psychosis Functional disturbances occur selectively in prefrontal cortex (thought and memory disorders) NMDA-Receptor antagonists (e.g. PCP) produce psychoses in healthy volunteers Schizophrenia may be related to altered glutamatergic neurotransmission Selective deficit of GABAergic transmission in prefrontal cortex

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Further hypotheses

Selective glutathion deficit (protection against oxidative stress) in prefrontal cortex Altered function of oligodendrocytes ( possible disturbance of myelination) Abnormal neuronal migration during formation of the neocortex (reelin hypothesis) Consequences of a prenatal production of inflammatory cytokines (disturbance of brain development during a critical period)

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Evidence for synaptic alterations in the prefrontal cortex of schizophrenia patients

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Altered regulation of dopaminergic function in schizophrenia

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Genotype-phenotype interactions in schizophrenia

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Allosteric modulation of GABAergic transmission by benzodiazepines

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Benzodiazepines
Benzodiazepines are drugs with a high affinity and selectivity for GABAA receptors. They differ mainly in their pharmacokinetic profile (half-life, active metabolites) Clinical applications of benzodiazepine agonists
Sleep disorders (sedation, hypnosis) Anxiety disorders (tranquillizer) Muscle spams, dystonia (muscle relaxant) Status epilepticus (anticonvulsants) Motor incoordination Anterograde amnesia Ethanol potentiation Tolerance
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Side effects

GABAA receptors

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Allosteric modulation by benzodiazepines

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GABAA receptor subtypes: pharmacology

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Diazepam-sensitive GABAA receptor subtypes

1

3 diazepam 5

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H101R: a molecular switch for diazepam sensitivity

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Wieland and Lddens, 1992

Diazepam-insensitive receptors in 1(H101R) mutants

3H

Ro15 4513 + diazepam

wt

Ro15 4513 is a ligand binding to all benzodiazepine sites; it can be displaced only from diazepamsensitive sites

3H

1(H101R)

1 subunit immunostaining

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Lack of sedative effect of diazepam in 1(H101R) mutants

WT
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1(H101R)

Further reading Books:

The biochemical basis of neuropharmacology (Cooper, Bloom, Roth) A primer of drug action (R.M. Julien; Freeman and company, New York) Molecular Neuropharmacology (E.J. Nestler, S. E. Hyman, R.C. Malenka; The McGraw-Hill Companies, Inc., New York)

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