Dopaminergic system function and dysfunction : experimental approaches

Experimental life sciences have two basic foundations: concepts and tools. The Neuromethods series focuses on the tools and techniques unique to the investigation of the nervous system and excitable cells. It will not, however, shortchange the concept side of things as care has been taken to integrate these tools within the context of the concepts and questions under investigation. In this way, the series is unique in that it not only collects protocols but also includes theoretical background information and critiques which led to the methods and their development. Thus, it gives the reader a better understanding of the origin of the techniques and their potential future development. The Neuromethods publishing program strikes a balance between recent and exciting developments like those concerning new animal models of disease, imaging, in vivo methods, and more established techniques, including, for example, immunocytochemistry and electrophysiological technologies. New trainees in neurosciences still need a sound footing in these older methods in order to apply a critical approach to their results.Under the guidance of its founders, Alan Boulton and Glen Baker, the Neuromethods series has been a success since its first volume published through Humana Press in 1985. The series continues to flourish through many changes over the years. It is now published under the umbrella of Springer Protocols. While methods involving brain research have changed a lot since the series started, the publishing environment and technology have changed even more radically. Neuromethods has the distinct layout and style of the Springer Protocols program, designed specifically for readability and ease of reference in a laboratory setting.The careful application of methods is potentially the most important step in the process of scientific inquiry. In the past, new methodologies led the way in developing new disciplines in the biological and medical sciences. For example, physiology emerged out of anatomy in the nineteenth century by harnessing new methods based on the newly discovered phenomenon of electricity. Nowadays, the relationships between disciplines and methods are more complex. Methods are now widely shared between disciplines and research areas. New developments in electronic publishing make it possible for scientists that encounter new methods to quickly find sources of information electronically. The design of individual volumes and chapters in this series takes this new access technology into account. Springer Protocols makes it possible to download single protocols separately. In addition, Springer makes its print-on-demand technology available globally. A print copy can therefore be acquired quickly and for a competitive price anywhere in the world. Preface to the Series Preface References Contents Contributors Part I: Neuroanatomy Chapter 1: Ultrastructural Study of Dopaminergic Axon Terminals 1 Introduction 2 Materials 2.1 Tissue Preparation 2.1.1 Perfusion-Fixation 2.1.2 Vibrating Microtome Sections 2.1.3 Freeze-Thaw 2.2 Immunohistochemistry 2.2.1 Primary and Secondary Antibody Incubation 2.2.2 Silver Intensification of Gold Conjugated to Secondary Antibody 2.3 Preparation of Sections for EM 2.3.1 Post-Fixation with Osmium Tetroxide 2.3.2 Dehydration and Embedding in Resin 2.3.3 Re-embedding for EM 2.3.4 Re-sectioning for EM 2.3.5 Staining of Ultrathin Sections with Reynold ́s Lead Citrate 2.4 Examination of Sections in the Electron Microscope 2.4.1 Image Acquisition 2.5 Analysis of Electron Micrographs 2.5.1 Software 3 Methods 3.1 Tissue Preparation 3.1.1 Perfusion-Fixation 3.1.2 Vibrating Microtome Sections 3.1.3 Freeze-Thaw 3.2 Immunohistochemistry 3.2.1 Primary and Secondary Antibody Incubation 3.2.2 Silver Intensification of Gold Conjugated to Secondary Antibody 3.3 Preparation of Sections for EM 3.3.1 Post-Fixation with Osmium Tetroxide 3.3.2 Dehydration and Embedding in Resin 3.3.3 Re-embedding for EM 3.3.4 Re-sectioning for EM 3.3.5 Staining of Ultrathin Sections with Reynold ́s Lead Citrate 3.4 Examination of Sections in the Electron Microscope 3.4.1 Sampling 3.4.2 Image Acquisition 3.5 Analysis of Electron Micrographs 3.5.1 Software 3.5.2 Image Analysis 4 Notes References Chapter 2: Quantifying Dopaminergic Innervation in Rodents Using Unbiased Stereology 1 Introduction 2 Materials 2.1 Tissue Processing 2.2 Stereology 3 Methods 3.1 Tissue Processing 3.1.1 Perfusion (See Note 2) 3.1.2 Tissue Sectioning and Storage 3.1.3 Tyrosine Hydroxylase (TH) Immunofluorescence 3.2 Stereology (See Note 8) 3.2.1 Calibration 3.2.2 Tracing the TH+ Innervation (See Note 13) 3.2.3 Using the Cavalieri Estimator to Calculate Region Volume 3.2.4 Optical Fractionator Workflow 3.2.5 Counting Objects (See Note 21) 3.2.6 Accounting for a Missing Section 3.2.7 OPTIONAL: Taking Image Stacks (See Note 22). 4 Notes References Chapter 3: Dendrites Crossing Borders: Quantifying the Distribution of Individual Substantia Nigra Dopaminergic Neurons ́ Dendr... 1 Introduction 2 Methods 2.1 Method 1: Manual Measurement of Dendrites in a Subdivision During Neuron Reconstruction 2.1.1 Method 1: Step by Step 2.2 Method 2: Manual Measurement of Dendrites in Subdivision Using a Rendered 3D Map 2.2.1 Methods 2: Step by Step 2.3 Method 3: Automatic Measurement of Dendrites Length in a Subdivision 2.3.1 Method 3: Step by Step 3 Conclusions and Future Perspectives References Part II: Cell Physiology Chapter 4: Characterizing Dopamine Transporter Mediated Uptake and Efflux in Brain Slices: A New Approach 1 Introduction 2 Materials 2.1 Obtaining Acute Rat Brain Slices 2.2 Rat Brain Superfusion Experiments for Measuring DA Efflux Using HPLC 2.3 High-Performance Liquid Chromatography HPLC to Measure DA Release 2.4 Multiwell-Based Assay for the Measurement of Endogenous DA Release Using HPLC 2.5 Use of Acute Brain Slices for Ex Vivo [3H]-DA Uptake Assay 3 Methods 3.1 Acute Rat Brain Slices for Both Uptake and Efflux Assays 3.2 Use of Acute Brain Slices for Ex Vivo [3H]-DA Uptake Assay 3.3 Rat Brain Superfusion for Measuring Endogenous Dopamine Efflux 3.4 Multiwell-Based Assay for the Measurement of Endogenous Dopamine Release 4 Notes 5 Discussion References Chapter 5: Modeling Pacemaking, Bursting, and Depolarization Block in Midbrain Dopamine Neurons 1 Introduction 2 Materials 3 Methods 3.1 Conductance-Based Models for Pacemaking 3.2 Phase Plane Analyses 3.3 Markov Models 3.4 Intrinsic Bursting 3.5 The Balanced State of Dopamine Neurons In Vivo 3.6 Biophysically Realistic Morphologies 4 Notes References Chapter 6: Electrophysiological Characterization of Dopaminergic Neurons of the Rat Substantia Nigra Compacta 1 Introduction 2 Materials and Drive Components 2.1 Wire Selection 2.2 Manipulator Assembly 2.3 Tube Array Soldering 2.4 Tetrode Twisting 2.5 Electrode Interfacing Using a Custom-Printed Circuit Board 3 Electrode Manipulator Assembly 3.1 Installing Cannulae Array Guide 3.2 Interfacing Each Single Wire of a Tetrode with PCB 3.3 Gold Plating 3.4 Improving Signal-to-Noise Ratio 4 Electrophysiological Setup 5 Qualitative and Quantitative Analysis of the Discharge Pattern of SNc Neurons 6 Classification of Dopaminergic Neurons and Effect of an AMPH Injection Bibliography Part III: Models for Addiction and Neuropsychiatric Diseases Chapter 7: Pulmonary Inhalation to Assess Effects of Coca Paste on Behavior and Dopamine Neurotransmission 1 Introduction 1.1 Cocaine Base Paste or Coca Paste: Definition 1.2 Coca Paste Chemical Composition: Cutting and Adulteration 1.3 Active Adulterant and Its Role in Cocaine Effects 1.4 Pulmonary Inhalation Route and Drug Addiction 2 Materials 2.1 Animals 2.2 Drugs 3 Methods 3.1 Procedure to Volatilize Drugs: Setup of Pulmonary Inhalation in Rats 3.2 Procedure to Study the Stimulant Effect of Drugs: Open Field Assay 3.3 Procedure to Study the Neurochemical Effect of Drugs: In Vivo Intracerebral Microdialysis Technique 4 Conclusions References Chapter 8: Ethanol Intake on Preclinical Models: Methods of Assessment and Effects on Dopamine Signaling 1 Introduction 1.1 Why Is It Important to Measure Ethanol Intake? How Preclinical Models Recapitulate Its Features? 1.2 Aim of the Chapter 1.3 Different Procedures to Model Ethanol Intake in Rodents 2 Assessing Ethanol Intake in Wistar Rats via an Adapted Version From the DID Paradigm: Materials 3 Assessing Ethanol Intake in Wistar Rats via an Adapted Version of the DID Paradigm: Procedure 4 Ethanol Self-Administration Modulates Dopaminergic System 5 DA Receptors, Agonists, and Antagonists 6 Concluding Comments 7 Notes References Chapter 9: Preclinical Models of THC and Nicotine Exposure During Adolescent Brain Development: Modeling Neuropsychiatric Phen... 1 Introduction 1.1 Drugs of Abuse and Adolescence 1.2 Evidence for the Impact of Neurodevelopmental Nicotine and Cannabinoid Exposure in Psychiatric Disorders 1.3 Rodent Models of Neurodevelopment: Similarities Between Human Vs. Rodent Brain Development Trajectories 2 Materials 2.1 Adolescent THC Treatment 2.2 Adolescent Nicotine Treatment 2.3 Behavioral Tests 2.3.1 Sucrose Preference 2.3.2 Social Interaction and Memory Test 2.3.3 Novel Object Recognition 2.4 Single Unit Recording 3 Methods 3.1 Adolescent THC Treatment 3.2 Adolescent Nicotine Treatment 3.3 Preclinical Behavioral Tests 3.3.1 Sucrose Preference 3.3.2 Social Interaction: Motivation and Memory 3.3.3 Novel Object Recognition 3.3.4 Software Analyses 3.4 Single-Unit Recording of Dopamine Neurons in VTA 3.4.1 Set up Multiclamp 700B Amplifier and Clampex Acquisition Software 3.4.2 Experimental Protocol 3.4.3 Results, Analyses, and Interpretations 4 Conclusions References Chapter 10: The MAM Model to Study the Role of Dopamine in Schizophrenia 1 Introduction 2 Material 2.1 MAM Compound 2.2 Timed Pregnant Rats 2.3 MAM Aliquoting Materials 2.4 Injection Materials 2.5 Bedding Changing Materials (Table 3) 2.6 Safety Concerns 3 Methods 3.1 Aliquoting MAM 3.2 Injection Procedure 3.3 Hazardous Period and Bedding Disposal 3.4 Disposal of MAM Materials 3.5 Postnatal Period 4 Discussion References Part IV: Circuit Level and Behavior Chapter 11: Fast Scan Cyclic Voltammetry to Assess Dopamine Function: From Circuits to Behavior 1 Introduction 2 Measuring Dopamine Activity in the Brain 2.1 Electrophysiology 2.2 Microdialysis 2.3 Fluorescent Biosensors 3 Fundamentals of Fast Scan Cyclic Voltammetry 3.1 Working and Reference Electrode Fabrication 3.2 Data Acquisition 3.2.1 Background Subtraction 3.2.2 Cycling of Electrodes 3.3 Data Analysis 3.3.1 Electrode Calibration 3.3.2 Analysis of Dopamine Release and Uptake 3.3.3 Chemometric Analysis 4 FSCV Variations and Applications 4.1 FSCV in Anesthetized Animals 4.1.1 Methods and Materials for FSCV in Anesthetized Animals 4.2 FSCV in Brain Slices 4.2.1 Methods and Materials for FSCV in Brain Slices 4.3 FSCV in Freely Moving Animals 4.3.1 Methods and Materials for FSCV in Awake, Freely Moving Animals 5 Effects of Psychostimulants on DA Neurotransmission 5.1 Acute Actions of Dopamine Transporter Inhibitors 5.2 Lasting Actions of Dopamine Transporter Inhibitors 6 Leveraging FSCV with Other Techniques 6.1 Electrophysiology 6.2 Biochemistry 6.3 Optogenetics and Chemogenetics 7 Conclusion References Chapter 12: Multi-circuit Recording in Animal Models of Parkinson's Disease 1 Introduction 2 Materials 2.1 Striatal Injection of 6-Hydroxydopamine 2.2 Electrode Implants 3 Methods 3.1 Striatal Injection of 6-Hydroxydopamine 3.2 Electrode Implants 4 Notes References Chapter 13: Neuroimaging Human Dopamine-Related Neurophysiology Across Development 1 Introduction 2 Overview of Iron in the Brain 3 Materials and Methods 3.1 MRI 3.2 Relaxometry-Based Approaches 3.2.1 R2 3.2.2 R2′ Methodological Considerations for R2* and R2′ 3.3 A Single-Echo Approximation of T2 Relaxation 3.3.1 Normalized T2*-Weighted Imaging 3.3.2 Methodological Considerations for nT2*-Weighted Imaging 3.4 Susceptibility-Based Approaches 3.4.1 Quantitative Susceptibility Mapping (QSM) 3.5 QSM Versus Relaxometry-Based Methods 3.6 Field Strength Considerations 4 Applications 4.1 Linking Tissue Iron to Striatal Dopamine 4.2 Linking Tissue Iron to Cognition Across the Lifespan 4.3 Linking Tissue Iron to Functional Changes in the Brain 5 Summary and Future Directions References Chapter 14: Dopaminergic Control of Actions and Habits 1 Introduction 1.1 Response Control by Actions and Habits 1.2 The Dopaminergic System and Its Role in Actions and Habits 1.3 Behavioral Tests to Assess Action Control 2 Materials 2.1 Animals 2.2 Housing 2.3 Behavioral Apparatus 2.4 Food Reward 3 Methods 3.1 Magazine Training 3.2 Instrumental Training 3.3 Outcome Devaluation 3.4 Contingency Degradation 3.5 Behavioral Analysis 3.6 Dopamine System, Action, and Habits: Future Directions 4 Notes References Index This volume provides a variety of technical approaches to study dopamine system function and dysfunction. Chapters guide readers through dopamine release in ex vivo and freely moving animals, multi-recording devices for in vivo simultaneous single cell and population activity, in silico modeling of dopamine neurons activity, neuroanatomical approaches, unbiased stereology, ultrastructural analyses of dopaminergic neurons, and axonal innervation. Additionally, chapters also incorporate pharmacological tools to model neuropsychiatric diseases, novel behavioral paradigms to dissect dopamine's role in behavior, and functional imaging to follow human dopamine system development. In the Neuromethods series style, chapters include the kind of detail and key advice from the specialists needed to get successful results in your laboratory. Comprehensive and cutting-edge, Dopamine Neurotransmission aims to be a valuable resource for researchers in various disciplines.