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Nicotinic Acetylcholine Receptors in Experimental Models of Parkinson's Disease and Levodopa-Induced Dyskinesia

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Nicotinic Acetylcholine Receptors in Experimental Models of Parkinson's Disease and Levodopa-Induced Dyskinesia Focus on α5 Subunit-Containing Receptors SAKARI LEINO Nicotinic Acetylcholine Receptors in Experimental Models of Parkinson’s Disease and Levodopa-Induced Dyskinesia: Recent Publications in this Series 92/2018 Jenni Puurunen Metabolomic Characterization of Canine Behavioural Disorders: Fearfulness and Hyperactivity/ Impulsivity 93/2018 Marit Ilves Immunomodulatory Effects of Engineered Nanomaterials in Healthy and Diseased Lungs and Skin DISSERTATIONES SCHOLAE DOCTORALIS AD SANITATEM INVESTIGANDAM 94/2018 Laura Lahdentausta UNIVERSITATIS HELSINKIENSIS 13/2019 Serum and Saliva Biomarkers in Cardiovascular Diseases and Periodontitis—Smoking as a Confounding Factor 95/2018 Joni Lindbohm Unfolding Lipid Profile- and Sex-Paradoxes in Epidemiology of Subarachnoid Haemorrhage 96/2018 Anna But Mitigating Bias and Dealing with Multiple Time Scales in Cohort Studies — Studying SAKARI LEINO Medications and Complications of Diabetes 97/2018 Ashwini S. 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I Supervisors: Docent Outi Salminen, Ph.D. Division of Pharmacology and Pharmacotherapy Faculty of Pharmacy University of Helsinki Finland Docent Saara Rannanpää, Ph.D. Division of Pharmacology and Pharmacotherapy Faculty of Pharmacy University of Helsinki Finland Reviewers: Professor Seppo Kaakkola, M.D., Ph.D. Clinicum Faculty of Medicine University of Helsinki Finland Professor Esa R. Korpi, M.D., Ph.D. Department of Pharmacology Medicum Faculty of Medicine University of Helsinki Finland Opponent: Professor Susan Wonnacott, Ph.D. Department of Biology & Biochemistry Faculty of Science University of Bath United Kingdom © Sakari Leino 2019 Dissertationes Scholae Doctoralis Ad Sanitatem Investigandam Universitatis Helsinkiensis ISBN 978-951-51-4854-4 (Paperback) ISBN 978-951-51-4855-1 (PDF) ISSN 2342-3161 (Paperback) ISSN 2342-317X (PDF) Helsinki, Finland 2019 Layout: Tinde Päivärinta/PSWFolders Oy/Ltd Press: Hansaprint Oy, Turenki II III Contents Abstract ..........................................................................................................................................VI Tiivistelmä ................................................................................................................................... VII Original publications ...............................................................................................................VIII Abbreviations ................................................................................................................................IX 1. Introduction ............................................................................................................................1 2. Review of the literature .........................................................................................................2 2.1. Basal ganglia and the nigrostriatal dopaminergic pathway ....................................2 2.1.1. Anatomy and neurocircuitry of the basal ganglia ......................................2 2.1.2. Dopaminergic modulation of basal ganglia neurotransmission ..............4 2.1.3. Striatal interneurons and non-dopaminergic neurotransmission ...........6 2.2. Parkinson’s disease and dopaminergic neurodegeneration ....................................8 2.2.1. Clinical features and epidemiology of Parkinson’s disease........................8 2.2.2. Pathophysiology of Parkinson’s disease .......................................................8 2.2.3. Current treatment of Parkinson’s disease and future perspectives .........13 2.3. Levodopa-induced dyskinesia ..................................................................................14 2.3.1. Clinical features and epidemiology ............................................................14 2.3.2. Pathophysiology of peak-dose LID ............................................................17 2.3.3. Current treatment options for LID and future perspectives ...................25 2.4. Nicotinic acetylcholine receptors in Parkinson’s disease .......................................26 2.4.1. Nicotinic receptor structure and function .................................................26 2.4.2. Nicotinic receptor-mediated neuromodulation in the basal ganglia .....30 2.4.3. α5* nicotinic receptors .................................................................................33 2.4.4. Nicotinic receptors as drug targets in Parkinson’s disease ......................35 3. Aims of the studies ...............................................................................................................47 4. Materials and methods ........................................................................................................48 4.1. Drugs ............................................................................................................................48 4.2. Animals ........................................................................................................................49 4.3. Stereotactic 6-OHDA administration and postoperative care .............................49 4.4. In vivo experiments ....................................................................................................50 4.4.1. Drug-induced rotametry and locomotor activity (Study I) ....................50 4.4.2. Drug treatments ............................................................................................50 4.4.3. Assessment of dyskinesia severity ..............................................................52 4.5. Immunohistochemistry .............................................................................................54 4.5.1. Tyrosine hydroxylase immunostaining ......................................................54 4.5.2. Densitometric and stereological analyses ..................................................54 4.6. Ex vivo assays ..............................................................................................................55 4.6.1. Synaptosomal [3H]dopamine uptake (Study I) .........................................55 4.6.2. Dopamine transporter Western blotting (Study I) ...................................56 4.6.3. Dopamine and metabolite HPLC (Study I) ...............................................56 4.6.4. Dopamine receptor qPCR (Study I) ...........................................................57 4.6.5. Synaptosomal [3H]dopamine release (Study II) .......................................57 4.6.6. BDNF ELISA (Study III) ..............................................................................58 4.7. Statistical methods ......................................................................................................59IV 5. Results .....................................................................................................................................60 5.1. Characterization of the 6-OHDA lesion models in wild-type mice ....................60 5.2. Eff ects of α5 gene deletion in mouse models of Parkinson’s disease ...................61 5.2.1. Eff ects of α5 gene deletion on dopaminergic denervation and drug-induced rotation (Study I) .................................................................61 5.2.2. Eff ects
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