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Table of contents Abbreviations ........................................................................................... 10 List of figures and tables .......................................................................... 13 Abstract ..............

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Table of contents

Abbreviations ... 10

List of figures and tables ... 13

Abstract ... 18

Introduction ... 20

1.The Striatum: the input nucleus of the Basal Ganglia system..……….……….22

1.1 The Striatum ... 22

1.1.1 Striatal neurons ... 22

1.1.1.1 Medium spiny neurons ... 22

1.1.1.2 Striatal interneurons ... 24

1.1.2 Striatal subregions ... 25

1.1.2.1 Dorsal striatum ... 26

1.1.2.1.1 Dorsomedial striatum ... 26

1.1.2.1.2 Dorsolateral striatum ... 27

1.1.2.1.3 Inputs to the dorsal striatum ... 29

1.1.2.1.4 Efferent pathways of the dorsal striatum ... 32

1.1.2.2 Nucleus Accumbens ... 33

1.1.2.2.1 General description ... 33

1.1.2.2.2 Inputs to the NAc ... 35

1.1.2.2.3 Efferent pathways ... 37

1.2 Output nuclei of the BG ... 38

1.2.1 Globus pallidus / ventral pallidum ... 38

1.2.2 Substantia nigra pars reticulata and pars compacta ... 39

1.2.3 Ventral tegmental area ... 39

1.2.4 Subthalamic nucleus ... 39

2. Decision-making behaviour ... 41

2.1 A brief introduction into decision-making theories and studies ... 41

2.2 Neural basis of decision-making ... 44

2.3 Behavioural tasks to evaluate decision-making processes ... 46

2.3.1 The Iowa Gambling Task (IGT) paradigm ... 47

2.3.2 Pavlovian-instrumental transfer (PIT)... 48

2.3.3 Effort-based decision-making protocol in a T-maze ... 50

Goals of the project ... 52

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1.Transgenic mouse strains ... 54

1.1 Drd1a-Cre mouse (D1-Cre) ... 54

1.2 Adora2a-R Cre mouse (A2A-Cre) ... 54

1.3 Rosa26-YFP mouse ... 55

1.4 Breeding ... 56

1.5 Housing conditions ... 57

1.6 Ethics ... 57

1.7 Genotyping ... 57

1.7.1 DNA extraction and PCR... 57

1.7.2 Agarose gel electrophoresis ... 58

2. Surgeries ... 58

2.1 Anesthesia ... 58

2.2 Stereotaxic surgery ... 59

2.2.1 Apparatus and general procedure ... 59

2.2.2 Virus use and delivery ... 60

2.2.3 Post-surgery recovery ... 60

2.3 Intracardiac perfusion with 4% paraformaldehyde (PFA) ... 61

3. Chemogenetics: Designer’s Receptor Exclusively Activated by Designer’s Drug (DREADD) approach ... 61

4. Behavioural procedures ... 64

4.1 Iowa Gambling Task (IGT)………64

4.1.1 Apparatus ... 64

4.1.2 Behavioural protocol ... 66

4.1.3 Pharmacological treatment ... 71

4.2 Open-field ... 71

4.3 Pavlovian-instrumental transfer (PIT) (adapted from Bertran-Gonzalez et al.,2013) ... 72

4.3.1 Behavioural protocol ... 72

4.4 T-maze for effort-based decision-making (adapted from Solinsky and Kirby,2013) ... 75

4.4.1 Behavioural protocol ... 75

5. Immunohistochemistry and image acquisition and processing ... 77

5.1 Brain slice preparation ... 77

5.2 C-fos immunostaining ... 77

5.3 GFP immunostaining ... 77

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6. Electrophysiological recordings ... 78

7. Statistical analysis ... 80

Results ... 83

1.Use of chemogenetics in the striatum ... 83

1.1 Transfection rates in A2A-YFP and D1-YFP transgenic mouse lines ... 83

1.2 Chemogenetic-dependent C-fos induction ... 83

1.3 Chemogenetic-dependent increase in locomotor activity in D1-Cre mice injected in the DMS ... 84

1.4 Testing of Addgene’s chemogenetic constructs ... 84

2. Behavioural paradigms to analyse decision-making ... 92

2.1 Pavlovian-instrumental transfer ... 92

2.2 T-maze: effort-based decision-making ... 95

2.3 The Iowa Gambling Task ... 97

3. Study of the involvement of the striatum in decision-making processes using the IGT: analysis of behavioural and neuronal modifications ... 101

3.1 Behavioural analysis ... 104

3.2 Electrophysiological recordings ... 127

4. C-fos immunostaining analysis to validate chemogenetic-mediated activation of targeted neurons after the IGT ... 132

Discussion ... 137

References ... 154

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