Genetic Dissection of Neural Circuits and Behavior (eBook)

Front Cover 1
Genetic Dissection of Neural Circuits and Behavior 4
Copyright Page 5
Contents 7
Contributors 9
Chapter 1: Genetic Dissection of Neural Circuits and Behavior in Mus musculus 11
Abstract 12
I. Conventional Gene Targeting Strategies 12
II. Spatially Restricted Gene Manipulation 15
A. The Cre/loxP system 16
B. Investigating NMDA receptor function in specific subregions of the brain using the Cre/loxP system 16
III. The Use of Dominant Negative Inhibitors to Manipulate Gene Function 20
IV. Conditional Manipulation of Gene Function 21
A. The tTA system 21
B. A novel form of the tTA system: The rtTA system 25
V. Pharmacogenetic Approaches 26
A. Tamoxifen-controlled gene manipulation 26
B. Pharmacogenetic regulation of neuronal excitability 28
C. Targeting of intracellular signaling pathways using pharmacogenetic approaches 29
VI. Optogenetic Approaches 32
VII. Visualization of Brain Structure and Function 34
A. The TetTag system 35
B. The Brainbow mouse 37
VIII. The Challenges Facing Genetic Approaches in the Mouse 38
Acknowledgments 41
References 42
Chapter 2: Worm Watching: Imaging Nervous System Structure and Function in Caenorhabditis elegans 49
I. Introduction 50
II. Real-Time Indicators of Neuronal Activity 53
A. Properties of worm neurons 53
B. Speed of electrical signaling 54
III. Monitoring Intracellular Calcium 55
A. GECIs: Cameleon and G-CaMP 55
B. Examples of GECIs in worm neurobiology 56
C. Limitations of GECIs 57
IV. Imaging Presynaptic Terminals 59
A. Quantifying presynaptic proteins 59
B. pH-Sensitive GFP at the synapse 61
C. Spatial localization as a signal 63
D. Systematic profiling of presynaptic proteins 63
V. Imaging Postsynaptic Compartments 64
A. Postsynaptic glutamate receptors 64
B. Postsynaptic nicotinic acetylcholine receptors 67
C. Other receptors 68
VI. Imaging Proxies for Neuropeptide Secretion 68
VII. Bimolecular Fluorescence Complementation in the Worm 70
A. Combinatorial promoters 70
B. GRASP 71
VIII. Methods for Immobilizing the Worm 71
A. Pharmacological paralysis 73
B. Glue 73
C. Microfluidics 74
D. Imaging on the go 74
IX. Femtosecond Lasers and Nanosurgery on the Worm 75
A. Beyond cell ablation 76
B. Axonal outgrowth and repair 76
X. Designing the Ideal Optical Probe 77
XI. Conclusions and Prospects 80
. Acknowledgments 80
. References 80
References 80
Chapter 3: Mapping and Manipulating Neural Circuits in the Fly Brain 89
. Abstract 90
I. Introduction 90
A. Genes for behavior 91
B. Neurons for behavior 93
C. Anatomy and stereotypy 94
II. Spatial Targeting of Neuron Types 96
III. Imaging Neurons 102
IV. Functional Imaging: Watching Neuronal Activity 105
A. Voltage sensors 106
B. Genetically encoded calcium indicators 107
V. Control of Neural Activity 110
A. Cell killers 113
B. Synaptic vesicle blockers 117
C. Electrical blockers 119
D. Neuronal activators 120
E. Light-based methods 121
F. Caveats 122
VI. Quantitative Behavioral Assays 123
VII. Conclusions 128
A. Example circuits 128
B. New tools 129
C. Full circle 130
. Acknowledgments 130
. References 131
References 131
Chapter 4: A Network of G-Protein Signaling Pathways Control Neuronal Activity in C. elegans 155
. Abstract 155
I. Introduction 156
A. Invertebrate neuroscience 156
B. C. elegans motorneurons 157
C. The C. elegans NMJ as a model synapse 158
D. Combining genetics with pharmacology 160
II. Mapping Intracellular Neural Networks Within C. Elegans Motorneurons 161
A. The EGL-30(Gaq) pathway 161
B. The GPA-12(Ga12) pathway 166
C. The GOA-1(Gao) pathway 167
D. The DAG effector pathways 169
E. The GSA-1(Gas) pathway 172
F. Other heterotrimeric G-protein subunits and regulators 177
G. G-protein-coupled receptors 178
H. Other genes that alter motorneuron activity 181
I. New screens-An embarrassment of riches 187
J. The C. elegans NMJ-A model for human synapses? 187
. References 188
References 188
Index 203
Color Plates 211