- A guide to running a behavioral testing lab, including the many aspects of mouse research beyond the confines of the specific test
- Diagrams and photographs are shown for many kinds of apparatus and test situations with sufficient details such as dimensions to enable building of replicas
- Provides step-by-step instructions on planning and executing behavioral experiments in order to run them successfully
Autor/Autorin: Douglas Wahlsten
Dr. Wahlsten received his Ph.D. degree in psychology from the University of California Irvine and completed his postdoctoral work in behavior genetics at the Institute for Behavioral Genetics at the University of Colorado. He then established research laboratories at the University of Waterloo, the University of Alberta, and the University of North Carolina. The laboratory work involved genetic and developmental analysis of mouse brain defects and methods of behavioral testing. He also acquired expertise in statistical data analysis and theoretical issues central to the field. He was a co-founder of the very successful journal Genes, Brain and Behavior, and was given the Distinguished Scientist award by the International Behavioral and Neural Genetics Society. Throughout his career, he has taught behavioral and neural genetics to undergraduate and graduate students.
Authoritative step-by-step guide to methods of mouse behavioral testing
Mouse Behavioral Testing: How to Use Mice in Behavioral Neuroscience provides detailed explanations of how to conduct an experiment on mouse behavior from the initial planning of the research design through every step of the process until the data analysis phase.
The book discusses the practical matters that need to be considered carefully when working with any species of animal, such as how many animals need to be tested. It describes the tests and techniques devised specifically for work with mice. Every step of the research process is illustrated with real situations encountered in previous studies. All examples are based on real experiments, and extensive details of several published experiments are provided. The essential features of a behavioral test protocol are outlined, and several complete protocols are provided. Methods to balance the order of tests and determine throughput are described, then a completely balanced order of tests in a complex experiment is presented.
The book will be useful for those already familiar with the general principles of research but are new to the realm of behavioral testing of live mice. It will also serve as a text for a formal course, most likely at the graduate level.
PrefaceAcknowledgmentsChapter 1 Introduction to the Research Process The Research Process: Scientific Aspects Adapting the approach to the stage of an investigation Mice, mice, and more mice Measuring instruments: Tests The formal research design Sample size Ethics approval Logistics Obtaining subjects The test day Prelude to data analysis Data analysis Publishing results The Institutional Context of Research People Facilities PoliciesChapter 2 Mice Mice Versus Rats Models of Human Functions Origins of Standard Mouse Strains Kinds of Breeding Schemes Outbred animals Closed colony Inbred strains F1 hybrids Selectively bred lines The Process of Close Inbreeding Inbred Strains for Research Special Genotypes Derived from Inbred Strains Coisogenic strains Congenic strains Consomic strains Conplastic strains Recombinant inbred strains Designer Mice: Transgenic Methods and Targeted MutationsChapter 3 Tests of Mouse Behavior Popularity of Tests Commercial Devices Exploration Open field Nose poke hole board Symmetrical Y maze Home cage activity Anxiety Lightdark box Elevated plus maze Elevated zero and square mazes Vogel conflict test Geller conflict test Motor Function Ataxia observation Grip strength Balance beam Grid test Rotarod Running wheel Treadmill Learning Operant learning Mazes Spatial memory Electric shock Other TestsChapter 4 Designs One Group Two Groups Matched Pairs Repeated Measures Single-Factor Study With More Than Two Groups (One-Way Design) Two-Factor Studies Multi-Factor Studies Specialized Designs Reciprocal hybrid crosses Manipulations of the maternal environment Transgenerational influences Selective breeding with control for environmental influences Complications Fixed and random effects Nested factors Litter as unit Consulting experts Utilities One- and two-factor studies Three- and higher factor studies Breeding listChapter 5 Sample Size Utilities to do the Calculations Population and Sample Two kinds of populations Specifying an entire population Random choice of a sample Comparing two groups Comparing several groups Size of an Effect What is an effect? Index of effect size Effect size for a study with two groups Effect size for more than two groups Finding effect size from published data Significance of an Effect False positives happen The more tests, the higher the risk of a false positive Power of a Test Finding the Correct Sample Size Two independent groups 2 2 design, 4 groups J groups, omnibus test J groups, linear contrast Two-way factorial design Designs with more than two factorsChapter 6 Ethics Approval Good Ethics and Good Science The Era Before Regulation The Era of Regulated Research With Animals Ethical Status of Mice and Rats The Fundamental Ethical Principle of Animal Research The 3Rs Replacement Reduction Refinement Case Studies Death as an end point Food deprivation Categories of Invasiveness and Severity Categories for Behavioral Tests Used with Mice The FutureChapter 7 Logistics The Total Experiment One-factor design Two-, three- and four-factor designs The Principle of Balancing and Randomization The Total Sample Divided into Smaller Units Throughput Partitioning the Work Day The Balancing Act Enumeration Latin square Randomization to the Rescue Shipments, Cages, Tail Marks, and Id Numbers Cages in the Colony Room The Data Sheet Final Consultation Other Examples More Intricate DesignsChapter 8 Getting Ready for Testing Ordering Mice Shipping Unpacking Marking for Individual Identification Housing Group versus individual housing Cage enrichment Lightdark cycle Going to School: Test DayChapter 9 Prelude to Data Analysis Know Your Object Contents of One Row of Data The Id Code Order of Variables Across a Row Naming Variables The Value of Using the Right Values The Plague Called Missing Data Importing the Spreadsheet into the Statistical Program Checking for Errors in the Data The Crucial Distinction Between Errors and Exceptions Looking at Results Statistical Data Analysis as a Final Step in the ProcessChapter 10 Domains and Test Batteries Types of Observations Whole behavior versus criterion response Free expression versus highly constrained actions Microscopic analysis versus the whole picture Short duration versus full day observation The Ethological Method Domains and Subdomains Complexity of Domains Test Batteries Every test changes the mouse Advantages of using different mice for different tests Efficiency and economy of numbers comes at a price Current wisdom about test batteries Testing effects of batteries Effect size and importance of test order effects One solution: Standard test orders How stressful are tests?Chapter 11 Motivating Mice Hunger and Food Deprivation Appetite without deprivation IntelliCage Methods of food deprivation A refined method for daily deprivation Degree of hunger and eating Revised eating test Electric Shock Basic features of a shock source Variables that influence intensity Determining the correct level of shock Threshold of sensation Threshold of overt reaction Kinds of responses and task requirements Water Escape Depth Temperature Number of trials Opacity Cleanliness Air Puff Aversion Parameters of air puffs Determining optimal air flow rate Avoidance of air puffs Motivation and LearningChapter 12 Qualities of Behavioral Tests Resolution, Accuracy, and Precision Consistency Reliability Replicability ValidityChapter 13 Task Refinement and Standardization Flawed Tests and Missteps in the Research Process The lab cat Mouse shuttle avoidance Bar press avoidance Task Complexity The relatively simple open field The submerged platform water escape task Do task parameters matter? Probe trial performance Escape latencies and rate of learning Task Refinement The accelerating rotarod The submerged platform water escape task Prospects for refining other tests Test Standardization Standardizing Lab EnvironmentChapter 14 Video Tracking Basic Features Camera and lens Spectral sensitivity and filters Focus of the lens Hardware Modifications for Video Tracking Solid floors for uniform fields of view Paint in the water tank Eliminating shadows and blind spots The experimenters hand Doubts about the Accuracy of Video Tracking Tracking in Edmonton and Portland VideoScan versus ANY-maze Accuracy and Precision from a Mechanical Device The Solution: Ir Backlighting The True Path LengthChapter 15 The Laboratory Environment Two Sources of Environmental Variance in Test Scores Environmental Effects on Test Scores: Early Studies Interactions with Lab Environment: Recent Studies Complexity of the Lab Environment Standardizing the Lab Environment Heterogenization of the Lab Environment Strategies for Standardizing the Lab EnvironmentAppendixReference ListIndex
Mouse Behavioral Testing: How to Use Mice in Behavioral Neuroscience provides detailed explanations of how to conduct an experiment on mouse behavior from the initial planning of the research design through every step of the process until the data analysis phase.
The book discusses the practical matters that need to be considered carefully when working with any species of animal, such as how many animals need to be tested. It describes the tests and techniques devised specifically for work with mice. Every step of the research process is illustrated with real situations encountered in previous studies. All examples are based on real experiments, and extensive details of several published experiments are provided. The essential features of a behavioral test protocol are outlined, and several complete protocols are provided. Methods to balance the order of tests and determine throughput are described, then a completely balanced order of tests in a complex experiment is presented.
The book will be useful for those already familiar with the general principles of research but are new to the realm of behavioral testing of live mice. It will also serve as a text for a formal course, most likely at the graduate level.
- A guide to running a behavioral testing lab, including the many aspects of mouse research beyond the confines of the specific test
- Diagrams and photographs are shown for many kinds of apparatus and test situations with sufficient details such as dimensions to enable building of replicas
- Provides step-by-step instructions on planning and executing behavioral experiments in order to run them successfully
Preface
Acknowledgments
Chapter 1 Introduction to the Research Process
The Research Process: Scientific Aspects
Adapting the approach to the stage of an investigation
Mice, mice, and more mice
Measuring instruments: Tests
The formal research design
Sample size
Ethics approval
Logistics
Obtaining subjects
The test day
Prelude to data analysis
Data analysis
Publishing results
The Institutional Context of Research
People
Facilities
Policies
Chapter 2 Mice
Mice Versus Rats
Models of Human Functions
Origins of Standard Mouse Strains
Kinds of Breeding Schemes
Outbred animals
Closed colony
Inbred strains
F1 hybrids
Selectively bred lines
The Process of Close Inbreeding
Inbred Strains for Research
Special Genotypes Derived from Inbred Strains
Coisogenic strains
Congenic strains
Consomic strains
Conplastic strains
Recombinant inbred strains
Designer Mice: Transgenic Methods and Targeted Mutations
Chapter 3 Tests of Mouse Behavior
Popularity of Tests
Commercial Devices
Exploration
Open field
Nose poke hole board
Symmetrical Y maze
Home cage activity
Anxiety
Lightdark box
Elevated plus maze
Elevated zero and square mazes
Vogel conflict test
Geller conflict test
Motor Function
Ataxia observation
Grip strength
Balance beam
Grid test
Rotarod
Running wheel
Treadmill
Learning
Operant learning
Mazes
Spatial memory
Electric shock
Other Tests
Chapter 4 Designs
One Group
Two Groups
Matched Pairs
Repeated Measures
Single-Factor Study With More Than Two Groups (One-Way Design)
Two-Factor Studies
Multi-Factor Studies
Specialized Designs
Reciprocal hybrid crosses
Manipulations of the maternal environment
Transgenerational influences
Selective breeding with control for environmental influences
Complications
Fixed and random effects
Nested factors
Litter as unit
Consulting experts
Utilities
One- and two-factor studies
Three- and higher factor studies
Breeding list
Chapter 5 Sample Size
Utilities to do the Calculations
Population and Sample
Two kinds of populations
Specifying an entire population
Random choice of a sample
Comparing two groups
Comparing several groups
Size of an Effect
What is an effect?
Index of effect size
Effect size for a study with two groups
Effect size for more than two groups
Finding effect size from published data
Significance of an Effect
False positives happen
The more tests, the higher the risk of a false positive
Power of a Test
Finding the Correct Sample Size
Two independent groups
2 2 design, 4 groups
J groups, omnibus test
J groups, linear contrast
Two-way factorial design
Designs with more than two factors
Chapter 6 Ethics Approval
Good Ethics and Good Science
The Era Before Regulation
The Era of Regulated Research With Animals
Ethical Status of Mice and Rats
The Fundamental Ethical Principle of Animal Research
The 3Rs
Replacement
Reduction
Refinement
Case Studies
Death as an end point
Food deprivation
Categories of Invasiveness and Severity
Categories for Behavioral Tests Used with Mice
The Future
Chapter 7 Logistics
The Total Experiment
One-factor design
Two-, three- and four-factor designs
The Principle of Balancing and Randomization
The Total Sample Divided into Smaller Units
Throughput
Partitioning the Work Day
The Balancing Act
Enumeration
Latin square
Randomization to the Rescue
Shipments, Cages, Tail Marks, and Id Numbers
Cages in the Colony Room
The Data Sheet
Final Consultation
Other Examples
More Intricate Designs
Chapter 8 Getting Ready for Testing
Ordering Mice
Shipping
Unpacking
Marking for Individual Identification
Housing
Group versus individual housing
Cage enrichment
Lightdark cycle
Going to School: Test Day
Chapter 9 Prelude to Data Analysis
Know Your Object
Contents of One Row of Data
The Id Code
Order of Variables Across a Row
Naming Variables
The Value of Using the Right Values
The Plague Called Missing Data
Importing the Spreadsheet into the Statistical Program
Checking for Errors in the Data
The Crucial Distinction Between Errors and Exceptions
Looking at Results
Statistical Data Analysis as a Final Step in the Process
Chapter 10 Domains and Test Batteries
Types of Observations
Whole behavior versus criterion response
Free expression versus highly constrained actions
Microscopic analysis versus the whole picture
Short duration versus full day observation
The Ethological Method
Domains and Subdomains
Complexity of Domains
Test Batteries
Every test changes the mouse
Advantages of using different mice for different tests
Efficiency and economy of numbers comes at a price
Current wisdom about test batteries
Testing effects of batteries
Effect size and importance of test order effects
One solution: Standard test orders
How stressful are tests?
Chapter 11 Motivating Mice
Hunger and Food Deprivation
Appetite without deprivation
IntelliCage
Methods of food deprivation
A refined method for daily deprivation
Degree of hunger and eating
Revised eating test
Electric Shock
Basic features of a shock source
Variables that influence intensity
Determining the correct level of shock
Threshold of sensation
Threshold of overt reaction
Kinds of responses and task requirements
Water Escape
Depth
Temperature
Number of trials
Opacity
Cleanliness
Air Puff Aversion
Parameters of air puffs
Determining optimal air flow rate
Avoidance of air puffs
Motivation and Learning
Chapter 12 Qualities of Behavioral Tests
Resolution, Accuracy, and Precision
Consistency
Reliability
Replicability
Validity
Chapter 13 Task Refinement and Standardization
Flawed Tests and Missteps in the Research Process
The lab cat
Mouse shuttle avoidance
Bar press avoidance
Task Complexity
The relatively simple open field
The submerged platform water escape task
Do task parameters matter?
Probe trial performance
Escape latencies and rate of learning
Task Refinement
The accelerating rotarod
The submerged platform water escape task
Prospects for refining other tests
Test Standardization
Standardizing Lab Environment
Chapter 14 Video Tracking
Basic Features
Camera and lens
Spectral sensitivity and filters
Focus of the lens
Hardware Modifications for Video Tracking
Solid floors for uniform fields of view
Paint in the water tank
Eliminating shadows and blind spots
The experimenters hand
Doubts about the Accuracy of Video Tracking
Tracking in Edmonton and Portland
VideoScan versus ANY-maze
Accuracy and Precision from a Mechanical Device
The Solution: Ir Backlighting
The True Path Length
Chapter 15 The Laboratory Environment
Two Sources of Environmental Variance in Test Scores
Environmental Effects on Test Scores: Early Studies
Interactions with Lab Environment: Recent Studies
Complexity of the Lab Environment
Standardizing the Lab Environment
Heterogenization of the Lab Environment
Strategies for Standardizing the Lab Environment
Appendix
Reference List
Index
Klappentext
This volume presents the essentials of behavioral test methodology needed for effective study planning and high quality behavioral data collection on laboratory mice. The broad scope of behavioral testing in relation to the neurosciences is discussed, as are the fine details of various test situations. Mistakes made in past research are scrutinized and lessons are drawn about how to avoid them in future work. Diverse factors that can influence the results of behavioral experiments are discussed, ranging from the specific parameters employed in a test to the laboratory environment outside the test situation itself. Throughout the book, emphasis is placed on special issues that arise when testing genetically different mice, including inbred strains and mice with targeted mutations. This knowledge will assist an investigator who lacks formal training in behavioral psychology to build a behavioral testing lab and insure that its output meets current standards of good practice in behavioral neuroscience research.
