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Drosophila Neurobiology, Second Edition


Subject Area(s):  Drosophila melanogaster; Laboratory Techniques; Neurobiology

Edited by Bing Zhang, University of Missouri; Ellie S. Heckscher, The University of Chicago; Alex C. Keene, Texas A&M University; Scott Waddell, University of Oxford

Due January 2025 • 930 pages (approx.), illustrated, index
Hardcover • $95 76.00
ISBN  978-1-621824-57-2
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  •     Description    
  •     Contents    

Description

Research on the fruit fly Drosophila melanogaster has played an important role in uncovering principles of nervous system structure and function. Fundamental insights include the first molecular descriptions of nervous system differentiation, the identification and elucidation of axon guidance cues, the first cloning of ion channels, the first demonstration that SNARE proteins are required for chemical neurotransmission, and the identification of single genes that regulate complex behavior. Early Drosophila neuroscience experiments used random mutagenesis to evaluate the genetic basis of complex traits. More recently, a rapidly expanding and more sophisticated genetic toolkit is readily available for precise genome modifications, and these approaches are complemented by protocols for studying neural function and behavior. Together, these methodologies have enabled and continue to drive significant progress in the field, from work investigating the earliest stages of nervous system development to the study of complex behaviors, including drug addiction and decision-making.

This second edition of the laboratory manual covers a comprehensive collection of review articles and protocols in Drosophila neuroscience. The articles are divided into three sections: neural development, neurophysiology, and behavior. The chapters discuss classic approaches as well as newer genetic techniques and behavioral paradigms. The contents of this manual are largely derived from those taught at the Cold Spring Harbor Laboratory “Drosophila Neurobiology” course, and the protocols are designed to be highly accessible for researchers at all levels and with diverse expertise and backgrounds. The breadth of the manual makes it not only an invaluable reference for researchers that currently work with these organisms, but also an essential start-up guide for those seeking to use Drosophila in their research for the first time.

Contents

Preface
SECTION 1: STUDYING NEURAL DEVELOPMENT IN DROSOPHILA
Chapter 1
INTRODUCTION
Studying Neural Development in Drosophila melanogaster: An Introduction to Section 1
Ellie S. Heckscher
Chapter 2
INTRODUCTION
Analysis of Axon Guidance in the Drosophila Embryo
Thomas Kidd and Timothy Evans
PROTOCOLS
1 Collection, Fixation, and Antibody Staining of Drosophila Embryos
Thomas Kidd and Timothy Evans
2 Ventral Nerve Cord Dissection and Microscopy of Drosophila Embryos
Thomas Kidd and Timothy Evans
Chapter 3
INTRODUCTION
Study of Axonal Injury and Degeneration in Drosophila
Thomas J. Waller, Laura J. Smithson, and Catherine A. Collins
PROTOCOLS
1 Wallerian Degeneration and Clearance of Olfactory Receptor Neuron Axons following Drosophila Antennal Transection
Thomas J. Waller, Laura J. Smithson, and Catherine A. Collins
2 Peripheral Nerve Crush in Drosophila Larvae
Thomas J. Waller, Laura J. Smithson, and Catherine A. Collins
3 Immobilizing Second-Instar Drosophila Larvae for Imaging and Surgery Using the Larva Chip
Laura J. Smithson, Thomas J. Waller, and Catherine A. Collins
4 Laser Microsurgery in Drosophila Larvae Using the MicroPoint Ablation System
Laura J. Smithson, Thomas J. Waller, and Catherine A. Collins
Chapter 4
INTRODUCTION
Study of Dendrite Differentiation Using Drosophila Dendritic Arborization Neurons
Jason Y. Tann, Fangke Xu, Minami Kimura, Oliver R. Wilkes, Li-Foong Yoong, Henrik Skibbe, and Adrian W. Moore
PROTOCOLS
1 Mosaic Analysis with a Repressible Cell Marker (MARCM) Clone Generation in Drosophila Dendritic Arborization Neurons
Jason Y. Tann, Fangke Xu, Minami Kimura, and Adrian W. Moore
2 Filleting and Immunostaining of Larvae to Visualize Drosophila Dendritic Arborization Neuron Dendrite Arbors
Fangke Xu, Jason Y. Tann, Oliver R. Wilkes, and Adrian W. Moore
3 Mounting of Embryos, Larvae, and Pupae for Live Drosophila Dendritic Arborization Neuron Imaging
Fangke Xu, Jason Y. Tann, Oliver R. Wilkes, Minami Kimura, Li-Foong Yoong, and Adrian W. Moore
4 Culture of Larval and Pupal Drosophila Dendritic Arborization Neurons
Jason Y. Tann, Fangke Xu, Li-Foong Yoong, and Adrian W. Moore
5 Use of DeTerm for Automated Drosophila Dendrite Arbor Terminal Counts
Minami Kimura, Jason Y. Tann, Oliver R. Wilkes, Fangke Xu, Henrik Skibbe, and Adrian W. Moore
Chapter 5
INTRODUCTION
Cell Biology Techniques for Studying Drosophila Peripheral Glial Cells
Katherine Clayworth, Mary Gilbert, and Vanessa Auld
PROTOCOLS
1 Dissection and Immunolabeling of the Central and Peripheral Nervous System of Drosophila Larvae
Katherine Clayworth, Mary Gilbert, and Vanessa Auld
2 Whole-Larva Cryosectioning and Immunolabeling of Drosophila Larvae
Katherine Clayworth, Mary Gilbert, and Vanessa Auld
3 Proximity Ligation Assay (PLA) for Fillets of Drosophila Larvae
Katherine Clayworth, Mary Gilbert, and Vanessa Auld
Chapter 6
INTRODUCTION
The Drosophila Larval Neuromuscular Junction: Developmental Overview
James Ashley and Robert A. Carrillo
PROTOCOL
1 Drosophila Late Embryonic through Late Larval Stage Body Wall Dissection: Dissection Tools and Techniques
James Ashley and Robert A. Carrillo
2 Immunohistochemistry and Morphometric Analysis of Drosophila Larval Body Wall Neuromuscular Junction Preparations
James Ashley and Robert A. Carrillo
3 Labeling of Cell Surface Proteins at the Drosophila Larval Neuromuscular Junction Using Binding Partner Peptides
James Ashley and Robert A. Carrillo
4 Using the Proximity Ligation Assay to Visualize Colocalization of Proteins at the Drosophila Larval Neuromuscular Junction
James Ashley and Robert A. Carrillo
5 Cell Ablation Techniques for the Larval Drosophila Neuromuscular Junction
James Ashley and Robert A. Carrillo
Chapter 7
INTRODUCTION
The Study of Sensorimotor Circuit Assembly in Drosophila melanogaster Embryos and Larvae
Chris C. Wreden and Ellie S. Heckscher
PROTOCOLS
1 Studying Drosophila Larval Behavior in Agarose Channels
Marie R. Greaney and Ellie S. Heckscher
2 Imaging Neural Activity in Intact, Semirestrained Drosophila Larvae
Deeptha Vasudevan, Chris C. Wreden, and Ellie S. Heckscher
3 Single-Neuron Labeling in Drosophila Using Multicolor FLP-Out
Zarion D. Marshall, Chris C. Wreden, and Ellie S. Heckscher
4 Fluorescent In Situ Hybridization Chain Reaction for RNA in the Drosophila Embryonic and Larval Central Nervous System
Jake E. Henderson, Chris C. Wreden, and Ellie S. Heckscher
Chapter 8
INTRODUCTION
Development of the Drosophila Optic Lobe
Yu Zhang and Xin Li
PROTOCOL
1 Immunostaining of Drosophila Brains to Study Optic Lobe Development
Yu Zhang and Xin Li
Chapter 9
INTRODUCTION
Developmental Genetic and Molecular Analysis of Drosophila Central Complex Lineages
Gonzalo N. Morales Chaya, Aisha Hamid, Adil R. Wani, Andrew Gutierrez, and Mubarak Hussain Syed
PROTOCOL
1 Lineage Analysis and Birthdating of Drosophila Central Complex Lineages
Adil R. Wani, Aisha Hamid, Gonzalo N. Morales Chaya, and Mubarak Hussain Syed
Chapter 10
INTRODUCTION
Using Chromatin Immunoprecipitation (ChIP) to Study the Chromatin State in Drosophila
Chengcheng Du and Pelin Volkan
PROTOCOLS
1 Sample Preparation for Chromatin Immunoprecipitation (ChIP) from Drosophila Antennal and Brain Samples
Chengcheng Du and Pelin Volkan
2 Chromatin Immunoprecipitation (ChIP) Using Drosophila Antennal and Brain Samples
Chengcheng Du and Pelin Volkan
3 Performing Quantitative PCR after Chromatin Immunoprecipitation (ChIP) of Drosophila Antennal and Brain Samples
Chengcheng Du and Pelin Volkan
Chapter 11
INTRODUCTION
Introduction to CRISPR and Its Use in Drosophila
Scott Gratz, Kate M. O’Connor-Giles, and Jill Wildonger
PROTOCOL
1 Generating CRISPR Alleles in Drosophila
Scott Gratz, Kate M. O’Connor-Giles, and Jill Wildonger
SECTION 2: STUDYING NEURONAL ACTIVITIES IN DROSOPHILA
Chapter 12
INTRODUCTION
Studying Neuronal Activities in Drosophila: An Introduction to Section 2
Bing Zhang
Chapter 13
INTRODUCTION
Synaptic Electrophysiology of the Drosophila Neuromuscular Junction
Bing Zhang and Bryan Stewart
PROTOCOLS
1 Electrophysiological Recording from a “Model” Cell
Bing Zhang and Bryan Stewart
2 Recording from Drosophila Larval Body Wall Muscles: Passive Membrane Properties and Basic Features of Synaptic Transmission
Bing Zhang and Bryan Stewart
3 Voltage-Clamp Analysis of Synaptic Transmission at the Drosophila Larval Neuromuscular Junction
Bing Zhang and Bryan Stewart
4 Focal Recording of Synaptic Currents from Single Boutons at the Drosophila Larval Neuromuscular Junction
Bing Zhang and Bryan Stewart
5 Fabrication of Microelectrodes, Suction Electrodes, and Focal Electrodes for Electrophysiological Recording in Drosophila
Bing Zhang and Bryan Stewart
Chapter 14
INTRODUCTION
Using Electrophysiology to Study Homeostatic Plasticity at the Drosophila Neuromuscular Junction
Tingting Wang and C. Andrew Frank
PROTOCOLS
1 Eliciting Presynaptic Homeostatic Potentiation at the Drosophila Larval Neuromuscular Junction
Tingting Wang and C. Andrew Frank
2 Measuring the Readily Releasable Synaptic Vesicle Pool at the Drosophila Larval Neuromuscular Junction
Tingting Wang and C. Andrew Frank
3 Measuring Presynaptic Calcium Influx at the Drosophila Larval Neuromuscular Junction
Tingting Wang and C. Andrew Frank
Chapter 15
INTRODUCTION
Patch-Clamping Fly Brain Neurons
Florencia Fernandez-Chiappe and Nara I. Muraro
PROTOCOLS
1 Preparation of Pipettes and Pipette-Filling Devices for Patch-Clamping Drosophila Neurons
Florencia Fernandez-Chiappe and Nara I. Muraro
2 Dissection of Drosophila Wandering Larval Brains for Patch-Clamping Neurons
Florencia Fernandez-Chiappe and Nara I. Muraro
3 Dissection of Drosophila Adult Brains for Patch-Clamping Neurons
Florencia Fernandez-Chiappe and Nara I. Muraro
4 Patch-Clamping Drosophila Brain Neurons
Florencia Fernandez-Chiappe and Nara I. Muraro
Chapter 16
INTRODUCTION
Whole-Brain Electrophysiology and Calcium Imaging in Drosophila during Sleep and Wake
Matthew Van De Poll, Lucy Tainton-Heap, Michael Troup, and Bruno van Swinderen
PROTOCOLS
1 Whole-Brain Electrophysiology in Drosophila during Sleep and Wake
Matthew Van De Poll and Bruno van Swinderen
2 Whole-Brain Calcium Imaging in Drosophila during Sleep and Wake
Lucy Tainton-Heap, Michael Troup, Matthew Van De Poll, and Bruno van Swinderen
Chapter 17
INTRODUCTION
Calcium Imaging of Neural Activity in Fly Photoreceptors
Pei-Ju Chen, Yan Li, and Chi-Hon Lee
PROTOCOLS
1 Dissection of the Head of a Live Fly for Functional Imaging
Pei-Ju Chen, Yan Li, and Chi-Hon Lee
2 Functional Imaging from Fly Photoreceptors
Pei-Ju Chen, Yan Li, and Chi-Hon Lee
3 Quantitative Analysis of Photoreceptor Intensity–Response Function in Fly Visual Neurons
Pei-Ju Chen, Yan Li, and Chi-Hon Lee
Chapter 18
INTRODUCTION
Chemosensory Coding in Drosophila Single Sensilla
Richard Benton and Anupama Dahanukar
PROTOCOLS
1 Recording from Fly Olfactory Sensilla
Richard Benton and Anupama Dahanukar
2 Recording from Fly Taste Sensilla
Anupama Dahanukar and Richard Benton
Chapter 19
INTRODUCTION
Imaging Neuropeptide Release from Drosophila Clock and Motor Neurons
Edwin S. Levitan, Dinara Bulgari, and Markus K. Klose
PROTOCOL
1 GFP and FAP Imaging of Neuropeptide Release in Drosophila
Edwin S. Levitan, Dinara Bulgari, and Markus K. Klose
Chapter 20
INTRODUCTION
Functional Imaging of Learning-Induced Plasticity in the Central Nervous System with Genetically Encoded Reporters in Drosophila
Tamara Boto and Seth M. Tomchik
PROTOCOLS
1 Imaging Olfactory Learning-Induced Plasticity In Vivo in the Drosophila Brain
Tamara Boto and Seth M. Tomchik
2 Ex Vivo Brain Imaging in Drosophila
Tamara Boto and Seth M. Tomchik
Chapter 21
INTRODUCTION
Transcriptional Genetically Encoded Calcium Indicators in Drosophila
Sonia Bansal and Suewei Lin
PROTOCOL
1 Imaging CaLexA and TRIC Signals in the Dissected Fly Central Nervous System
Sonia Bansal and Suewei Lin
Chapter 22
INTRODUCTION
Analysis of Mitochondrial Dynamics in Adult Drosophila Axons
Daniel C. Maddison, Francesca Mattedi, Alessio Vagnoni, and Gaynor Ann Smith
PROTOCOLS
1 Clonal Imaging of Mitochondria in the Dissected Fly Wing
Daniel C. Maddison, Francesca Mattedi, Alessio Vagnoni, and Gaynor Ann Smith
2 Live Imaging of Mitochondria in the Intact Fly Wing
Francesca Mattedi, Daniel C. Maddison, Gaynor Ann Smith, and Alessio Vagnoni
SECTION 3: STUDYING BEHAVIOR IN DROSOPHILA
Chapter 23
INTRODUCTION
Studying Behavior in Drosophila: An Introduction to Section 3
Scott Waddell and Alex C. Keene
Chapter 24
INTRODUCTION
Neuronal Regulation of Energy Homeostasis in Drosophila: A Practical Approach
Tânia Reis
PROTOCOL
1 Density Assay for Body Fat Determination in Drosophila Larvae
Tânia Reis
Chapter 25
INTRODUCTION
Learning and Memory in Drosophila Larvae
Denise Weber, Vincent Richter, Astrid Rohwedder, Alexandra Großjohann, and Andreas S. Thum
PROTOCOL
1 The Analysis of Aversive Olfactory–Taste Learning and Memory in Drosophila Larvae
Denise Weber, Vincent Richter, Astrid Rohwedder, Alexandra Großjohann, and Andreas S. Thum
Chapter 26
INTRODUCTION
Innate and Conditioned Taste Processing in Drosophila
Elizabeth Brown, Kurtresha Worden, Yuanyuan Li, Pavel Masek, and Alex C. Keene
PROTOCOLS
1 Measurement of Reflexive Feeding Response in Drosophila
Elizabeth Brown, Kurtresha Worden, Yuanyuan Li, Pavel Masek, and Alex C. Keene
2 Measurement of Taste Memory in Drosophila
Elizabeth Brown, Kurtresha Worden, Yuanyuan Li, Pavel Masek, and Alex C. Keene
Chapter 27
INTRODUCTION
Measurement of Drosophila Reproductive Behaviors
Anne C. von Philipsborn, Galit Shohat-Ophir, and Carolina Rezaval
PROTOCOLS
1 Single-Pair Courtship and Competition Assays in Drosophila
Anne C. von Philipsborn, Galit Shohat-Ophir, and Carolina Rezaval
2 Courtship Conditioning/Suppression Assays in Drosophila
Anne C. von Philipsborn, Galit Shohat-Ophir, and Carolina Rezaval
3 Probing Acoustic Communication during Fly Reproductive Behaviors
Anne C. von Philipsborn, Galit Shohat-Ophir, and Carolina Rezaval
4 Female Fly Postmating Behaviors
Anne C. von Philipsborn, Galit Shohat-Ophir, and Carolina Rezaval
Chapter 28
INTRODUCTION
Ethanol Behavioral Responses in Drosophila
Kavin M. Nuñez, Jamie L. Catalano, Kristin M. Scaplen, and Karla R. Kaun
PROTOCOL
1 Methods for Exploring the Circuit Basis of Ethanol-Induced Changes in Drosophila Group Locomotor Activity
Kavin M. Nuñez, Jamie L. Catalano, Kristin M. Scaplen, and Karla R. Kaun
Chapter 29
INTRODUCTION
Nociception in Drosophila Larvae
Lydia J. Borjon, Stephanie E. Mauthner, and W. Daniel Tracey
PROTOCOLS
1 Mechanical Nociception Assay in Drosophila Larvae
Stephanie E. Mauthner and W. Daniel Tracey
2 Optogenetic Stimulation of Nociceptive Escape Behaviors in Drosophila Larvae
Stephanie E. Mauthner and W. Daniel Tracey
3 Assaying Nociception Behaviors in Drosophila Larvae during Parasitoid Wasp Attacks
Stephanie E. Mauthner, Lydia J. Borjon, and W. Daniel Tracey
Chapter 30
INTRODUCTION
Tracking the Navigation Behavior of Drosophila Larvae in Real and Virtual Odor Gradients by Using the Raspberry Pi Virtual Reality (PiVR) System
David Tadres, Nitesh Saxena, and Matthieu Louis
PROTOCOL
1 Using the Raspberry Pi Virtual Reality (PiVR) System to Study Drosophila Larval Chemotaxis with Real and Virtual Odor Gradients
David Tadres, Nitesh Saxena, and Matthieu Louis
Chapter 31
INTRODUCTION
Twists to Classical Conditioning of Adult Drosophila
Zeynep Okray and Scott Waddell
PROTOCOL
1 Classical Conditioning of Adult Drosophila
Zeynep Okray, Pedro F. Jacob, John-Paul Moszynski, Clifford B. Talbot, and Scott Waddell
Chapter 32
INTRODUCTION
Fighting Flies: Quantifying and Analyzing Drosophila Aggression
Maria P. Fernandez, Severine Trannoy, and Sarah J. Certel
PROTOCOL
1 Comparing Methods for Quantifying and Analyzing Drosophila Aggression
Severine Trannoy, Maria P. Fernandez, and Sarah J. Certel
Chapter 33
INTRODUCTION
Activity Monitoring for Analysis of Sleep in Drosophila melanogaster
Divya Sitaraman, Christopher G. Vecsey, and Casey Koochagian
PROTOCOLS
1 Neural Stimulation during Drosophila Activity Monitor (DAM)-Based Studies of Sleep and Circadian Rhythms in Drosophila melanogaster
Christopher G. Vecsey, Casey Koochagian, Martin Reyes, and Divya Sitaraman
2 Analysis of Positional Preference in Drosophila Using Multibeam Activity Monitors
Maria T. Porter, Gregg Roman, and Christopher G. Vecsey
3 Analysis of Sleep and Circadian Rhythms from Drosophila Activity-Monitoring Data Using SCAMP
Christopher G. Vecsey, Casey Koochagian, Maria T. Porter, Gregg Roman, and Divya Sitaraman
Appendix: General Safety and Hazardous Material Information
Index