Current Protocols in Neuroscience

Current Protocols in Neuroscience

Online ISBN: 9780471142300

DOI: 10.1002/0471142301

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  1. Preface
  2. Foreword
  3. Selected Suppliers of Reagents and Equipment
  4. Chapter 1 Neuroanatomical Methods
    1. Introduction
    2. UNIT 1.1 Basic Neuroanatomical Methods
    3. UNIT 1.2 Immunohistochemical Localization of Proteins in the Nervous System
    4. UNIT 1.3 Hybridization Histochemistry of Neural Transcripts
    5. UNIT 1.4 Receptor Binding Techniques
    6. UNIT 1.5 Transneuronal Circuit Analysis with Pseudorabies Viruses
    7. UNIT 1.6 Metabolic Mapping
    8. UNIT 1.7 Targeted Toxins
    9. UNIT 1.8 Cellular Compartment Analysis of Temporal Activity by Fluorescence In Situ Hybridization (catFISH)
    10. UNIT 1.9 Detection of DNA Damage in Tissue Sections by In Situ Nick Translation
    11. UNIT 1.10 Quantitative In Situ Hybridization for the Study of Gene Expression at the Regional and Cellular Levels
    12. UNIT 1.11 Counting Cells in Sectioned Material: A Suite of Techniques, Tools, and Tips
    13. UNIT 1.12 Intracellular and Juxtacellular Staining with Biocytin
    14. UNIT 1.13 Quantified Assessment of Terminal Density and Innervation
    15. UNIT 1.14 Anterograde Axonal Tract Tracing
    16. UNIT 1.15 Histochemical Methods for the Detection of Apoptosis in the Nervous System
    17. UNIT 1.16 Biochemical Fractionation of Brain Tissue for Studies of Receptor Distribution and Trafficking
    18. UNIT 1.17 Retrograde Axonal Tracing with Fluorescent Markers
    19. UNIT 1.18 Evolution of Neurotoxins: From Research Modalities to Clinical Realities
    20. UNIT 1.19 Mapping Lineage Using BAC-Cre Reporter Lines
    21. UNIT 1.20 Adeno-Associated Viral Vectors for Anterograde Axonal Tracing with Fluorescent Proteins in Nontransgenic and Cre Driver Mice
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      UNIT 1.21 How to Prepare Neuroanatomical Image Data
    23. UNIT 1.22 Non-isotopic RNA In Situ Hybridization on Embryonic Sections
    24. Unit 1.23 Fixation and Immunolabeling of Brain Slices: SNAPSHOT Method
    25. UNIT 1.24 Cre Activated and Inactivated Recombinant Adeno-Associated Viral Vectors for Neuronal Anatomical Tracing or Activity Manipulation
    26. UNIT 1.25 Whole Mouse Brain Image Reconstruction from Serial Coronal Sections Using FIJI (ImageJ)
    27. UNIT 1.26 Anterograde or Retrograde Transsynaptic Circuit Tracing in Vertebrates with Vesicular Stomatitis Virus Vectors
  5. Chapter 2 Imaging
    1. Introduction
    2. UNIT 2.1 Fluorescence Microscopy: A Concise Guide to Current Imaging Methods
    3. UNIT 2.2 Basic Confocal Microscopy
    4. UNIT 2.3 Imaging Nervous System Activity
    5. UNIT 2.4 Electronic Imaging in Neuroscience
    6. UNIT 2.5 Loading Neurons with Dextran-Conjugated Calcium Indicators in Intact Nervous Tissue
    7. UNIT 2.6 Use of FM1-43 and Other Derivatives to Investigate Neuronal Function
    8. UNIT 2.7 Characterizing Synaptic Vesicle Proteins Using Synaptosomal Fractions and Cultured Hippocampal Neurons
    9. UNIT 2.8 3-D Reconstruction of Neurons from Multichannel Confocal Laser Scanning Image Series
    10. UNIT 2.9 Two-Photon Imaging in Live Rodents
    11. UNIT 2.10 Whole-Mount Immunohistochemistry of the Brain
    12. UNIT 2.11 Ballistic Labeling with Fluorescent Dyes and Indicators
    13. UNIT 2.12 The Importance of Titrating Antibodies for Immunocytochemical Methods
    14. UNIT 2.13 DiOlistic Labeling in Fixed Brain Slices: Phenotype, Morphology, and Dendritic Spines
    15. UNIT 2.14 High-Speed Multineuron Calcium Imaging Using Nipkow-Type Confocal Microscopy
    16. UNIT 2.15 Confocal Laser Scanning Microscopic Photoconversion: A New Method to Stabilize Fluorescently Labeled Cellular Elements for Electron Microscopic Analysis
    17. UNIT 2.16 Use of Channelrhodopsin for Activation of CNS Neurons
    18. UNIT 2.17 Fluorescence Recovery After Photobleaching (FRAP) with a Focus on F-actin
    19. UNIT 2.18 Tracking Quantum Dot–Tagged Calcium Channels at Vertebrate Photoreceptor Synapses: Retinal Slices and Dissociated Cells
    20. UNIT 2.19 Assembly of Synapses: Biomimetic Assays to Control Neurexin/Neuroligin Interactions at the Neuronal Surface
    21. UNIT 2.20 Single-Molecule Tracking Photoactivated Localization Microscopy to Map Nano-Scale Structure and Dynamics in Living Spines
    22. UNIT 2.21 Imaging mGluR5 Dynamics in Astrocytes Using Quantum Dots
    23. UNIT 2.22 Optical Clearing of Fixed Brain Samples Using SeeDB
    24. UNIT 2.23 Time-Lapse Imaging of Glial–Axonal Interactions
    25. UNIT 2.24 Laser-Mediated Microlesions in Mouse Neocortex to Investigate Neuronal Degeneration and Regeneration
    26. UNIT 2.25 The Membrane Marker mCLING Reveals the Molecular Composition of Trafficking Organelles
  6. Chapter 3 Cellular and Developmental Neuroscience
    1. Introduction
    2. UNIT 3.1 Culture of Neuroepithelial Stem Cells
    3. UNIT 3.2 Long-Term Culture of Hippocampal Neurons
    4. UNIT 3.3 Culture of Substantia Nigra Neurons
    5. UNIT 3.4 Isolation and Purification of Primary Oligodendrocyte Precursors
    6. UNIT 3.5 Isolation and Purification of Primary Rodent Astrocytes
    7. UNIT 3.6 Differentiation of Embryonic Stem Cells
    8. UNIT 3.7 Immortalizing Central Nervous System Cells
    9. UNIT 3.8 Identification of Neural Programmed Cell Death Through the Detection of DNA Fragmentation In Situ and by PCR
    10. UNIT 3.9 Quantitative Analysis of In Vivo Cell Proliferation
    11. UNIT 3.10 Neural Stem Cell Transplantation in Mouse Brain
    12. UNIT 3.11 Calcium Phosphate Transfection of DNA into Neurons in Primary Culture
    13. UNIT 3.12 Purification of Oligodendrocytes and Their Progenitors Using Immunomagnetic Separation and Percoll Gradient Centrifugation
    14. UNIT 3.13 Isolation and Generation of Oligodendrocytes by Immunopanning
    15. UNIT 3.14 Generation and Utilization of Phosphorylation State–Specific Antibodies to Investigate Signaling Pathways
    16. UNIT 3.15 Neuronal Transfection Using Particle-Mediated Gene Transfer
    17. UNIT 3.16 Gene Expression in Transgenic Mice Using Neural Promoters
    18. UNIT 3.17 Isolation and Purification of Primary Schwann Cells
    19. UNIT 3.18 Fluorescence-Based Sorting of Neural Stem Cells and Progenitors
    20. UNIT 3.19 Rodent Sensory Neuron Culture and Analysis
    21. UNIT 3.20 Dissection and Culture of Embryonic Spinal Commissural Neurons
    22. UNIT 3.21 Culture of Postnatal Mesencephalic Dopamine Neurons on an Astrocyte Monolayer
    23. UNIT 3.22 A Method for Preparing Primary Retinal Cell Cultures for Evaluating the Neuroprotective and Neuritogenic Effect of Factors on Axotomized Mature CNS Neurons
    24. UNIT 3.23 In Vitro Analysis of Mouse Mesencephalic Neural Crest Development
    25. UNIT 3.24 Culture of Mouse Olfactory Sensory Neurons
    26. UNIT 3.25 Culturing Embryonic Nasal Explants for Developmental and Physiological Study
    27. UNIT 3.26 Culturing of Cerebellar Granule Cells to Study Neuronal Migration: Gradient and Local Perfusion Assays
    28. UNIT 3.27 Primary Porcine Brain Microvessel Endothelial Cell Isolation and Culture
    29. UNIT 3.28 Tracking Neuronal Migration in Adult Brain Slices
    30. UNIT 3.29 Real-time Recordings of Migrating Cortical Neurons from GFP and Cre Recombinase Expressing Mice
  7. Chapter 4 Gene Cloning, Expression, and Mutagenesis
    1. Introduction
    2. UNIT 4.1 Homology Cloning of cDNA or Genomic DNA
    3. UNIT 4.2 PCR Cloning of Neural Gene Products
    4. UNIT 4.3 Expression Cloning of Neural Genes Using Xenopus laevis Oocytes
    5. UNIT 4.4 Interaction Trap/Two-Hybrid System to Identify Interacting Proteins
    6. UNIT 4.5 Overview of Neural Gene Expression
    7. UNIT 4.6 Selection of Transfected Mammalian Cells
    8. UNIT 4.7 Transient Expression of Proteins Using COS Cells
    9. UNIT 4.8 cDNA Expression Cloning in Mammalian Cells
    10. UNIT 4.9 Mutagenesis Approaches for Elucidation of Protein Structure-Function Relationships
    11. UNIT 4.10 Oligonucleotide-Directed Mutagenesis Without Phenotypic Selection
    12. UNIT 4.11 Directed Mutagenesis Using the Polymerase Chain Reaction
    13. UNIT 4.12 Overview of Gene Delivery into Cells Using HSV-1-Based Vectors
    14. UNIT 4.13 Generation of High-Titer Defective HSV-1 Vectors
    15. UNIT 4.14 Gene Delivery Using Helper Virus-Free HSV-1 Amplicon Vectors
    16. UNIT 4.15 The Substituted-Cysteine Accessibility Method (SCAM) to Elucidate Membrane Protein Structure
    17. UNIT 4.16 Protein Expression in the Drosophila Schneider 2 Cell System
    18. UNIT 4.17 Production of Recombinant Adeno-Associated Viral Vectors and Use in In Vitro and In Vivo Administration
    19. UNIT 4.18 Overview of the Baculovirus Expression System
    20. UNIT 4.19 Protein Expression in the Baculovirus System
    21. UNIT 4.20 Calcium Flux Assay in Xenopus Oocytes
    22. UNIT 4.21 Production and Titration of Lentiviral Vectors
    23. UNIT 4.22 Alphaviruses: Semliki Forest Virus and Sindbis Virus Vectors for Gene Transfer into Neurons
    24. UNIT 4.23 Gene Transfer into Neural Cells In Vitro Using Adenoviral Vectors
    25. UNIT 4.24 Gene Transfer into Rat Brain Using Adenoviral Vectors
    26. UNIT 4.25 Overview of Nucleic Acid Arrays
    27. UNIT 4.26 Preparation of mRNA for Expression Monitoring
    28. UNIT 4.27 cDNA Library Construction from Single Cells
    29. UNIT 4.28 Gene Expression Analysis of Neural Cells and Tissues Using DNA Microarrays
    30. UNIT 4.29 Overview of Gene Targeting by Homologous Recombination
    31. UNIT 4.30 Production of a Heterozygous Mutant Cell Line by Homologous Recombination (Single Knockout)
    32. UNIT 4.31 Conditional Gene Expression and Targeting in Neuroscience Research
    33. UNIT 4.32 Transfection of Cultured Primary Neurons via Nucleofection
    34. UNIT 4.33 Generation of Designer Receptors Exclusively Activated by Designer Drugs (DREADDs) Using Directed Molecular Evolution
    35. UNIT 4.34 Transfection of Mouse Cochlear Explants by Electroporation
    36. UNIT 4.35 Manipulating Gene Expression in Projection-Specific Neuronal Populations Using Combinatorial Viral Approaches
    37. UNIT 4.36 The Use of Herpes Simplex Virus in Ex Vivo Slice Culture
    38. UNIT 4.37 Use of Adeno-Associated and Herpes Simplex Viral Vectors for In Vivo Neuronal Expression in Mice
  8. Chapter 5 Molecular Neuroscience
    1. Introduction
    2. UNIT 5.1 RNA Analysis by Nuclease Protection
    3. UNIT 5.2 Differential Display of mRNA by PCR
    4. UNIT 5.3 Analysis of mRNA Populations from Single Live and Fixed Cells of the Central Nervous System
    5. UNIT 5.4 Reducing Gene Expression in the Brain via Antisense Methods
    6. UNIT 5.5 Production of Polyclonal Antisera
    7. UNIT 5.6 Production of Antipeptide Antisera
    8. UNIT 5.7 Production of Antisera Using Fusion Proteins
    9. UNIT 5.8 Epitope Tagging of Recombinant Proteins
    10. UNIT 5.9 Overview of Membrane Protein Solubilization
    11. UNIT 5.10 Metal-Chelate Affinity Chromatography
    12. UNIT 5.11 Immunoaffinity Chromatography
    13. UNIT 5.12 The Use of Phage Display in Neurobiology
    14. UNIT 5.13 Fluorescent In Situ Transcription in Cells and Tissues
    15. UNIT 5.14 Detection of Protein Phosphorylation in Tissues and Cells
    16. UNIT 5.15 Use of the A. Victoria Green Fluorescent Protein to Study Protein Dynamics In Vivo
    17. UNIT 5.16 Green Fluorescent Protein in the Study of Neuronal Signaling Pathways
    18. UNIT 5.17 Analysis of RNA by Northern and Slot-Blot Hybridization
    19. UNIT 5.18 Using Phage Display in Neurobiology
    20. UNIT 5.19 Immunoblotting and Immunodetection
    21. UNIT 5.20 An Overview on the Generation of BAC Transgenic Mice for Neuroscience Research
    22. UNIT 5.21 Modification of Bacterial Artificial Chromosomes (BACs) and Preparation of Intact BAC DNA for Generation of Transgenic Mice
    23. UNIT 5.22 Quantifying Cellular Dynamics by Fluorescence Resonance Energy Transfer (FRET) Microscopy
    24. UNIT 5.23 Monitoring Protein-Protein Interactions in Living Cells by Bioluminescence Resonance Energy Transfer (BRET)
    25. UNIT 5.24 Immunoprecipitation
    26. UNIT 5.25 Detection of Protein-Protein Interactions by Coprecipitation
    27. UNIT 5.26 Lentiviral Strategies for RNAi Knockdown of Neuronal Genes
    28. UNIT 5.27 Split-Ubiquitin System for Identifying Protein-Protein Interactions in Membrane and Full-Length Proteins
    29. UNIT 5.28 Identifying Novel Protein-Protein Interactions Using Co-Immunoprecipitation and Mass Spectroscopy
    30. UNIT 5.29 Visualization of G Protein−Coupled Receptor (GPCR) Interactions in Living Cells Using Bimolecular Fluorescence Complementation (BiFC)
    31. UNIT 5.30 A Protein Cross-Linking Assay for Measuring Cell Surface Expression of Glutamate Receptor Subunits in the Rodent Brain After In Vivo Treatments
    32. UNIT 5.31 Imaging the Insertion of Superecliptic pHluorin-Labeled Dopamine D2 Receptor Using Total Internal Reflection Fluorescence Microscopy
  9. Chapter 6 Neurophysiology
    1. Introduction
    2. UNIT 6.1 The Electrophysiology Setup
    3. UNIT 6.2 Principles of Signal Conditioning
    4. UNIT 6.3 Fabrication of Patch Pipets
    5. UNIT 6.4 Preparation of Hippocampal Brain Slices
    6. UNIT 6.5 Acute Isolation of Neurons from the Mature Mammalian Central Nervous System
    7. UNIT 6.6 Whole-Cell Voltage Clamp Recording
    8. UNIT 6.7 Patch-Pipet Recording in Brain Slices
    9. UNIT 6.8 Single-Channel Recording
    10. UNIT 6.9 Rapid Solution Application Methods
    11. UNIT 6.10 Recording and Analyzing Synaptic Currents and Synaptic Potentials
    12. UNIT 6.11 Preparation and Maintenance of Organotypic Slice Cultures of CNS Tissue
    13. UNIT 6.12 Voltage Clamp Recordings from Xenopus Oocytes
    14. UNIT 6.13 Synaptic Plasticity in the Hippocampal Slice Preparation
    15. UNIT 6.14 Amperometry and Cyclic Voltammetry with Carbon Fiber Microelectrodes at Single Cells
    16. UNIT 6.15 Preparation and Maintenance of Organotypic Cultures for Multi-Electrode Array Recordings
    17. UNIT 6.16 Chronic Recording of Extracellular Neuronal Activity in Behaving Animals
    18. UNIT 6.17 Imaging Nervous System Activity with Voltage-Sensitive Dyes
    19. UNIT 6.18 Recording in the Cerebellar Slice
    20. UNIT 6.19 Patch-Clamp Recording from Neuronal Dendrites
    21. UNIT 6.20 Caged Molecules: Principles and Practical Considerations
    22. UNIT 6.21 Photolysis of Caged Neurotransmitters: Theory and Procedures for Light Delivery
    23. UNIT 6.22 Whole-Cell Recording In Vivo
    24. UNIT 6.23 Electrophysiological Recordings from Neonatal Neocortical Brain Slices
    25. UNIT 6.24 EEG Recording in Rodents, with a Focus on Epilepsy
    26. UNIT 6.25 Electroencephalography (EEG) and Event-Related Potentials (ERPs) with Human Participants
    27. UNIT 6.26 Electrophysiology Tool Construction
    28. UNIT 6.27 Incubation of Fear
    29. UNIT 6.28 Quantifying Recording Quality in In Vivo Striatal Recordings
    30. UNIT 6.29 Electrophysiological Measurement of Cannabinoid-Mediated Synaptic Modulation in Acute Mouse Brain Slices
  10. Chapter 7 Neurochemistry/Neuropharmacology
    1. Introduction
    2. UNIT 7.1 Overview of Brain Microdialysis
    3. UNIT 7.2 Microdialysis in Rodents
    4. UNIT 7.3 Microdialysis in Nonhuman Primates
    5. UNIT 7.4 Detection and Quantification of Neurotransmitters in Dialysates
    6. UNIT 7.5 Analyzing Binding Data
    7. UNIT 7.6 Saturation Assays of Radioligand Binding to Receptors and Their Allosteric Modulatory Sites
    8. UNIT 7.7 Saturation Analysis of Ligand Binding Using a Centrifugation Procedure
    9. UNIT 7.8 Ligand Characterization Using Microphysiometry
    10. UNIT 7.9 Uptake and Release of Neurotransmitters
    11. UNIT 7.10 Measurement of Chloride Movement in Neuronal Preparations
    12. UNIT 7.11 Measurement of Cation Movement in Primary Cultures Using Fluorescent Dyes
    13. UNIT 7.12 Measurement of Second Messengers in Signal Transduction: cAMP and Inositol Phosphates
    14. UNIT 7.13 Measurement of NO and NO Synthase
    15. UNIT 7.14 Measurement of Nitric Oxide in Single Cells and Tissue Using a Porphyrinic Microsensor
    16. UNIT 7.15 Scintillation Proximity Assay
    17. UNIT 7.16 Peripheral Type Benzodiazepine Binding Sites as a Tool for the Detection and Quantification of CNS Injury
    18. UNIT 7.17 Measurement of Oxygen Radicals and Lipid Peroxidation in Neural Tissues
    19. UNIT 7.18 Assessment of Cell Viability in Primary Neuronal Cultures
    20. UNIT 7.19 In Vivo Measurement of Blood-Brain Barrier Permeability
    21. UNIT 7.20 Methods for Sample Preparation for Direct Immunoassay Measurement of Analytes in Tissue Homogenates: ELISA Assay of Amyloid β-Peptides
    22. UNIT 7.21 Assays for Cyclic Nucleotide-Specific Phosphodiesterases (PDEs) in the Central Nervous System (PDE1, PDE2, PDE4, and PDE10)
    23. UNIT 7.22 Isolation of Mitochondria from the CNS
    24. UNIT 7.23 High-Affinity Choline Uptake (HACU) and Choline Acetyltransferase (ChAT) Activity in Neuronal Cultures for Mechanistic and Drug Discovery Studies
    25. UNIT 7.24 Direct Bisulfite Sequencing for Examination of DNA Methylation with Gene and Nucleotide Resolution from Brain Tissues
    26. UNIT 7.25 Sampling Phasic Dopamine Signaling with Fast-Scan Cyclic Voltammetry in Awake, Behaving Rats
  11. Chapter 8 Behavioral Neuroscience
    1. Introduction
    2. UNIT 8.1 Locomotor Behavior
    3. UNIT 8.2 Sexual and Reproductive Behaviors
    4. UNIT 8.3 Animal Tests of Anxiety
    5. UNIT 8.4 Application of Experimental Stressors in Laboratory Rodents
    6. UNIT 8.5A Assessment of Spatial Memory Using the Radial Arm Maze and Morris Water Maze
    7. UNIT 8.5B Assessment of Spatial Memory Using the T Maze
    8. UNIT 8.5C Cued and Contextual Fear Conditioning in Mice
    9. UNIT 8.5D Social Learning of Food Preferences in Rodents: Rapid Appetitive Learning
    10. UNIT 8.5E Assessment of Sustained and Divided Attention in Rats
    11. UNIT 8.5F Assessment of Learning and Memory Using the Autoshaping of Operant Responding in Mice
    12. UNIT 8.5G Social Transmission of Food Preference in Mice
    13. UNIT 8.5H Assaying Aspects of Attention and Impulse Control in Mice Using the 5-Choice Serial Reaction Time Task
    14. UNIT 8.5I Social Recognition Assay in the Rat
    15. UNIT 8.6A Overview of Methodological Approaches to the Study of Ingestive Behavior
    16. UNIT 8.6B Basic Measures of Food Intake
    17. UNIT 8.6C Direct, Continuous Behavioral Analysis of Drug Action on Feeding
    18. UNIT 8.6D Sham Feeding in Rats with Chronic, Reversible Gastric Fistulas
    19. UNIT 8.6E Conditioned Flavor Aversions: Assessment of Drug-Induced Suppression of Food Intake
    20. UNIT 8.6F Conditioned Flavor Preferences: Evaluating Postingestive Reinforcement by Nutrients
    21. UNIT 8.6G Macronutrient Selection in Experimental Animals
    22. UNIT 8.7 Measurement of Startle Response, Prepulse Inhibition, and Habituation
    23. UNIT 8.8 Measurement of Rodent Stereotyped Behavior
    24. UNIT 8.9 Models of Nociception: Hot-Plate, Tail-Flick, and Formalin Tests in Rodents
    25. UNIT 8.10A Rodent Models of Depression: Forced Swim and Tail Suspension Behavioral Despair Tests in Rats and Mice
    26. UNIT 8.10B Rodent Models of Depression: Learned Helplessness Using a Triadic Design in Rats
    27. UNIT 8.10C Rodent Models of Depression: Learned Helplessness Induced in Mice
    28. UNIT 8.11A Fear-Potentiated Startle in Rats
    29. UNIT 8.11B Fear-Potentiated Startle in Mice
    30. UNIT 8.12 Motor Coordination and Balance in Rodents
    31. UNIT 8.13 Latent Inhibition
    32. UNIT 8.14 Ultrasonic Vocalizations in Rat and Mouse Pups
    33. UNIT 8.15 Parental Behaviors in Rats and Mice
    34. UNIT 8.16 Mouse Social Recognition and Preference
    35. UNIT 8.17 Assessment of Murine Startle Reactivity, Prepulse Inhibition, and Habituation
    36. UNIT 8.18 Assessment of Developmental Milestones in Rodents
    37. UNIT 8.19 Defensive Responses to Predator Threat in the Rat and Mouse
    38. UNIT 8.20 Olfactometry with Mice
    39. UNIT 8.21A Overview of Methods for Assessing the Mouse Auditory System
    40. UNIT 8.21B Measurement of the Auditory Brainstem Response (ABR) to Study Auditory Sensitivity in Mice
    41. UNIT 8.21C Assessment of Cochlear Function in Mice: Distortion-Product Otoacoustic Emissions
    42. UNIT 8.21D Behavioral Assessment of Hearing in Mice—Conditioned Suppression
    43. UNIT 8.22 Delay Discounting and Impulsive Choice in the Rat
    44. UNIT 8.23 Fear Extinction in Rodents
    45. UNIT 8.24 Simple Behavioral Assessment of Mouse Olfaction
    46. UNIT 8.25 Modeling Appetitive Pavlovian-Instrumental Interactions in Mice
    47. UNIT 8.26 Automated Three-Chambered Social Approach Task for Mice
    48. UNIT 8.27 A Mouse Model for Observational Fear Learning and the Empathetic Response
    49. UNIT 8.28 Analysis of Skilled Forelimb Movement in Rats: The Single Pellet Reaching Test and Staircase Test
    50. UNIT 8.29 Methods for Studying Habitual Behavior in Mice
    51. UNIT 8.30 Assessing Delay Discounting in Mice
    52. UNIT 8.31 Examining Object Location and Object Recognition Memory in Mice
    53. UNIT 8.32 Avoidance Behavior: A Free-Operant Lever-Press Avoidance Task for the Assessment of the Effects of Safety Signals
    54. UNIT 8.33 To Stress or Not to Stress: A Question of Models
    55. UNIT 8.34 Measuring Motivation and Reward-Related Decision Making in the Rodent Operant Touchscreen System
  12. Chapter 8 Behavioral Neroscience
    1. UNIT 8.35 Studying Socio-Affective Communication in Rats through Playback of Ultrasonic Vocalizations
  13. Chapter 9 Preclinical Models of Neurologic and Psychiatric Disorders
    1. Introduction
    2. UNIT 9.1 Dural Inflammation Model of Migraine Pain
    3. UNIT 9.2 Traumatic Brain Injury in the Rat Using the Fluid-Percussion Model
    4. UNIT 9.3 Preclinical Models of Hepatic Encephalopathy
    5. UNIT 9.4 Preclinical Models of Parkinson's Disease
    6. UNIT 9.5 Rodent Models of Global Cerebral Ischemia
    7. UNIT 9.6 Rodent Models of Focal Cerebral Ischemia
    8. UNIT 9.7 Experimental Autoimmune Encephalomyelitis (EAE)
    9. UNIT 9.8 A Murine Model of Retroviral Encephalopathy
    10. UNIT 9.9 Animal Models of Retroviral Encephalopathies: Feline Model
    11. UNIT 9.10 Overview of Rodent Models for Obesity Research
    12. UNIT 9.11 Intravenous Self-Administration of Ethanol in Mice
    13. UNIT 9.12 Operant Self-Administration Models for Testing the Neuropharmacological Basis of Ethanol Consumption in Rats
    14. UNIT 9.13 Models of Amyotrophic Lateral Sclerosis
    15. UNIT 9.14 Models of Neuropathic Pain in the Rat
    16. UNIT 9.15 Place Preference Test in Rodents
    17. UNIT 9.16 Inducing Photochemical Cortical Lesions in Rat Brain
    18. UNIT 9.17 Measurement of Panic-Like Responses Following Intravenous Infusion of Sodium Lactate in Panic-Prone Rats
    19. UNIT 9.18 Animal Models of Painful Diabetic Neuropathy: The STZ Rat Model
    20. UNIT 9.19 Chemoconvulsant Model of Chronic Spontaneous Seizures
    21. UNIT 9.20 Chronic Intravenous Drug Self-Administration in Rats and Mice
    22. UNIT 9.21 Intravenous Self-Administration Techniques in Monkeys
    23. UNIT 9.22 Overview on Rodent Models of Alzheimer's Disease
    24. UNIT 9.23A Binge Eating in Rats Produced by Combining Dieting with Stress
    25. UNIT 9.23B Binge Eating in Rats with Limited Access to Vegetable Shortening
    26. UNIT 9.23C Sugar Bingeing in Rats
    27. UNIT 9.23D Characterization of Energy Expenditure in Rodents by Indirect Calorimetry
    28. UNIT 9.24 Overview of Animal Models of Schizophrenia
    29. UNIT 9.25 Ratings of L-DOPA-Induced Dyskinesia in the Unilateral 6-OHDA Lesion Model of Parkinson's Disease in Rats and Mice
    30. UNIT 9.26 Overview of Mouse Assays of Ethanol Intoxication
    31. UNIT 9.27 Schedule-Induced Polydipsia: A Rat Model of Obsessive-Compulsive Disorder
    32. UNIT 9.28 The Majchrowicz Binge Alcohol Protocol: An Intubation Technique to Study Alcohol Dependence in Rats
    33. UNIT 9.29 Vapor Inhalation of Alcohol in Rats
    34. UNIT 9.30 Animal Models of Obsessive-Compulsive Disorder
    35. UNIT 9.31 An Animal Model of Premenstrual Dysphoric Disorder Sensitive to Antidepressants
    36. UNIT 9.32 Recapitulation and Reversal of a Persistent Depression-like Syndrome in Rodents
    37. UNIT 9.33 A Murine Model of Atypical Antipsychotic-Induced Weight Gain and Metabolic Dysregulation
    38. UNIT 9.34 Drug-Induced Sexual Dysfunction in Rats
    39. UNIT 9.35 Overview of Animal Models of Attention Deficit Hyperactivity Disorder (ADHD)
    40. UNIT 9.36 Open-Space Forced Swim Model of Depression for Mice
    41. UNIT 9.37 Pentylenetetrazol (PTZ) Kindling Model of Epilepsy
    42. UNIT 9.38 Diet-Induced Models of Obesity (DIO) in Rodents
    43. UNIT 9.39 Prime-, Stress-, and Cue-Induced Reinstatement of Extinguished Drug-Reinforced Responding in Rats: Cocaine as the Prototypical Drug of Abuse
    44. UNIT 9.40 Rodent Models of Colorectal Distension
    45. UNIT 9.41 Blast Traumatic Brain Injury in the Rat Using a Blast Overpressure Model
    46. UNIT 9.42 Social Fear Conditioning as an Animal Model of Social Anxiety Disorder
    47. UNIT 9.43 The MAM Rodent Model of Schizophrenia
    48. UNIT 9.44 A Choice-Based Screening Method for Compulsive Drug Users in Rats
    49. UNIT 9.45 Animal Models of Post-Traumatic Stress Disorder
    50. UNIT 9.46 Modeling an Inflammation-Related Depressive Phenotype in Mice Using Bacille Calmette-Guérin
    51. UNIT 9.47 Rodent Model of Activity-Based Anorexia
    52. UNIT 9.48 The 3-NP Model of Striatal Neurodegeneration
    53. UNIT 9.49 “Drinking in the Dark” (DID): A Simple Mouse Model of Binge-Like Alcohol Intake
    54. UNIT 9.50 Latent Sensitization: A Model for Stress-Sensitive Chronic Pain
    55. UNIT 9.51 Attentional Set-Shifting Paradigm in the Rat
    56. UNIT 9.52 The Revised Neurobehavioral Severity Scale (NSS-R) for Rodents
  14. Chapter 10 Human Neuroscience
    1. Introduction
    2. UNIT 10.1 Assessment of Parkinson Disease Manifestations
    3. UNIT 10.2 EEG Recording and Analysis for Sleep Research
    4. UNIT 10.3 Neuropsychological Assessment in the Diagnosis of Alzheimer's Disease
    5. UNIT 10.4 Clinical and Neuroimaging Assessments for Research Studies (Including Drug Trials) in Multiple Sclerosis
    6. UNIT 10.5 A Rubric for Extracting Idea Density from Oral Language Samples
  15. Appendix 1 Molecular Biology Techniques
    1. APPENDIX 1A Molecular Biology References
    2. APPENDIX 1B Optimization of Transfection
    3. APPENDIX 1C Calcium Phosphate Transfection
    4. APPENDIX 1D Transfection Using DEAE-Dextran
    5. APPENDIX 1E Transfection by Electroporation
    6. APPENDIX 1F Transfection of Cultured Eukaryotic Cells Using Cationic Lipid Reagents
    7. APPENDIX 1G Purification and Concentration of DNA from Aqueous Solutions
    8. APPENDIX 1H Preparation of Genomic DNA from Mammalian Tissue
    9. APPENDIX 1I Preparation of RNA from Tissues and Cells
    10. APPENDIX 1J Preparation of Bacterial Plasmid DNA
    11. APPENDIX 1K Quantitation of DNA and RNA with Absorption and Fluorescence Spectroscopy
    12. APPENDIX 1L Introduction of Plasmid DNA into Cells
    13. APPENDIX 1M Digestion of DNA with Restriction Endonucleases
    14. APPENDIX 1N Agarose Gel Electrophoresis
  16. Appendix 2 Stock Solutions, Equipment, and Laboratory Guidelines
    1. APPENDIX 2A Common Stock Solutions, Buffers, and Media
    2. APPENDIX 2B Standard Laboratory Equipment
  17. Appendix 3 Commonly Used Techniques
    1. APPENDIX 3A Preparation of an Iodinated Radioligand
    2. APPENDIX 3B Techniques for Mammalian Cell Tissue Culture
  18. Appendix 4 Animal Techniques
    1. APPENDIX 4A High Precision Stereotaxic Surgery in Mice
    2. APPENDIX 4B Mouse and Rat Anesthesia and Analgesia
    3. APPENDIX 4C Animal Health Assurance
    4. APPENDIX 4D Handling and Restraint
    5. APPENDIX 4E Animal Identification
    6. APPENDIX 4F Parenteral Injections
    7. APPENDIX 4G Blood Collection
    8. APPENDIX 4H Euthanasia
    9. APPENDIX 4I Assessing Reproductive Status/Stages in Mice
  19. Appendix 5 Databases
    1. APPENDIX 5A Internet Resources for Genomic, Bioinformatics, and Medical Genetics Information
    2. APPENDIX 5B Databases and Web Sites for Neurogenetics