Advances in Metabolic Mapping Techniques for Brain Imaging of Behavioral and Learning Functions: Proceedings of the NATO Advanced Research Workshop, Austin, Texas, U.S.A., November 7-9, 1991(v. 68 NATO Science Series D:)

In recent years, revolutionary technical advances have permitted neuroscientists to map the functioning of the brain in exquisite detail. Of interest are the new techniques that visually display cell energy metabolism which is coupled to functional brain activity in behaving animals. This is the first book dealing with the application of 2-deoxyglucose and related metabolic mapping techniques for brain imaging of behavioral and learning functions. Quantitative autoradiographic techniques based on the use of exogenous markers include radiolabeled glucose and its analogs, especially 2-deoxyglucose and fluorodeoxyglucose. Other mapping techniques are based on the histochemical staining of endogenous metabolic markers such as cytochrome oxidase, as well as immunohistochemistry for expression of c-fos genes. In spite of the great potential capabilities of the new imaging techniques, relatively few neuroscientists are using this approach to study brain functions related to behavior. There is a need to review state-of-the-art applications of these methods in behavioral neuroscience, and to formulate recommendations for future research in this area. This book is intended to fulfill these needs by bringing together leading neuroscientists using metabolic mapping approaches to elucidate brain mechanisms of behavior. Discussions are not limited to one animal species, but they cover a broad range of vertebrates with unique behavioral capabilities.

Table of Contents:
I: Techniques.- Imaging Techniques in Studies of Neural Functions.- 1. Introduction.- 2. Local Cerebral Blood Flow.- 3. Local Cerebral Glucose Utilization.- 3.1. Theoretical Basis of Radioactive Deoxyglucose Method.- 3.2. Considerations in Design of Procedure.- 3.3. Experimental Procedure.- 3.4. Local Rates of Cerebral Glucose Utilization.- 3.5. The [18F]Fluorodeoxyglucose Technique.- 3.6. Metabolic Mapping of Local Functional Activity.- 4. Local Cerebral Protein Synthesis.- 4.1. Theory.- 4.2. Determination of ?i.- 4.3. Local Rates of Protein Synthesis in the Conscious Rat Brain.- 4.4. Applications of Autoradiographic L-[1-14C]Leucine Method.- 5. Miscellaneous Imaging Methods.- 6. References.- Brain Imaging of Auditory Learning Functions in Rats: Studies with Fluorodeoxyglucose Autoradiography and Cytochrome Oxidase Histochemistry.- 1. Introduction.- 1.1. Brain Imaging with Metabolic Mapping Techniques.- 1.2. Neuronal Activity in Sensory Systems is Modified by Learning.- 1.3. Metabolic Mapping Studies show Learning-Related Modifications in the Activity of the Auditory System.- 2. Principles of Fluorodeoxyglucose Autoradiography and their Applications for Neuroimaging of Learning Functions.- 2.1. Goals for FDG Autoradiographic Experiments.- 2.2. Transport and Uptake of FDG as a Glucose Analog.- 2.3. Tracer Specific Activity and Testing Conditions.- 2.4. Tissue Processing for FDG Autoradiography.- 2.5. Quantitative Analysis of FDG Autoradiographs.- 2.6. Procedure for FDG Quantitative Autoradiography.- 3. Application of FDG to the Study of Behavioral Habituation to an Auditory Stimulus.- 3.1. Short and Long-Term Habituation of the Acoustic Startle Reflex.- 3.2. Experimental Design and Methods.- 3.3. Effects on the Auditory System.- 3.4. Effects Outside the Auditory System.- 3.5. Model of the Neural Circuitry Related to Long-Term Habituation of the Acoustic Startle Reflex.- 4. Application of FDG to the Study of Differential Conditioning of Auditory Stimuli.- 4.1. Neural Effects of Sounds Differentially Associated with Appetitive and Aversive Unconditioned Stimuli.- 4.2. Experimental Design and Methods.- 4.3. Effects on the Auditory System.- 4.4. Effects Outside the Auditory System.- 4.5. Model of the Neural Circuitry Related to Differential Conditioning of Auditory Stimuli.- 5. Principles of Cytochrome Oxidase Histochemistry and their Applications for Neuroimaging of Learning Functions.- 5.1. Goals for CO Histochemical Experiments.- 5.2. The use of CO as an Endogenous Marker for Neuronal Function.- 5.3. CO Enzymology and Quantitative Histochemistry.- 5.4. Procedure for CO Quantitative Histochemistry.- 6. Application of CO Histochemistry to the Study of Learning Functions.- 6.1. Interassay Variability and Linearity for Group Comparisons.- 6.2. Learning-Related Increase in Metabolic Capacity.- 7. Conclusions.- 7.1. Application of FDG Autoradiography to Long-Term Habituation Revealed the Functional Brain Circuitry Mediating this Simple Form of Learning.- 7.2. Application of FDG Autoradiography to Differential Conditioning Revealed the Functional Brain Circuitry Mediating this Form of Discrimination Learning.- 7.3. Application of CO Histochemistry Revealed Long-Lasting Modifications in Metabolic Capacity Related to Chronic Learning.- 7.4. Learning-Related Changes are Distributed in Neural Systems with Specific Functional Contributions to Modify Behavior.- 8. Acknowledgements.- 9. References.- Mapping Sensorimotor Pathways in Rat Brain Using 2-Deoxyglucose Autoradiography and C-Fos Immunocytochemistry.- 1. Introduction-2-DG Studies of Thalamus.- 2. 2-DG Methods-Whisker Stimulation Following Cortical Lesions.- 3. Conclusions of 2-DG Studies.- 4. Introduction- c-fos Studies.- 5. Methods-Cortical Stimulation with c-fos and 2-DG.- 6. Results- c-fos and 2-DG.- 7. Conclusions.- 8. Comments on Quantitation of 2-DG and c-fos Studies.- 9. Acknowledgements.- 10. References.- Brain Metabolic Mapping and Behavior: Assessing the Effects of Early Developmental Experiences in Adult Animals.- 1. General Introduction.- 1.1. General Methodological Aspects.- 2. Study I: 14C-2-DG Autoradiography Under Normal vs. Challenge Conditions: Effects of Neonatal 6-OHDA Lesions and Rearing Environment.- 2.1. Methods.- 2.2. Results.- 2.3. Discussion.- 3. Study II: Effects of Prenatal Cocaine Exposure on Regional Brain Glucose Metabolism and Cytochrome Oxidase Histochemistry.- 3.1. Methods.- 3.2. Results.- 3.3. Discussion.- 4. General Conclusions.- 5. Acknowledgements.- 6. References.- High Resolution Autoradiographic Imaging of Brain Activity Patterns With 2-Deoxyglucose: Regional Topographic and Cellular Analysis.- 1. Introduction.- 2. Methods and Examples of Results.- 2.1. Animal Preparation for Injection of 2-DG.- 2.2. Improvement of Regional Topographic Resolution in 2-DG Studies.- 2.3. Cellular Resolution in 2-DG Studies.- 3. Consideration of the Time Course of 2-DG Studies.- 4. Quantitative Considerations in 5 Minutes 2-DG Studies.- 5. Summary and Conclusions.- 6. Acknowledgements.- 7. References.- Metabolic Mapping in the Hippocampus. Patterns of (14C)2-Deoxyglucose Uptake Within Different Fields of the Hippocampal Slice.- 1. Introduction.- 2. Material and Methods.- 2.1. Tissue Preparation.- 2.2. Electrophysiology.- 2.3. [14C]2-Deoxyglucose Incubation.- 2.4. Potassium Stimulation.- 2.5. Histology.- 2.6. Densitometric Analysis.- 3. Results.- 3.1. Comparison with Previous in vivo Results.- 3.2. Potassium Stimulation.- 4. Discussion.- 4.1. Potassium Stimulation.- 5. Acknowledgements.- 6. References.- Covariance Analysis of Functional Interactions in the Brain Using Metabolic and Blood Flow Data.- 1. Introduction.- 2. Methods.- 3. Examples of Correlational Analyses.- 3.1. Nucleus Basalis Magnocellularis Lesions in the Rat.- 3.2. Obsessive-Compulsive Disorder.- 3.3. Object and Spatial Vision in Humans.- 4. What are the Neurobiological Substrates ofInterregional Correlations?.- 5. Brain Network Modeling of Metabolic Data.- 5.1. The Model.- 5.2. Examples of Simulations.- 5.3. Data-Fitting and Combinations of Neural Models.- 6. Acknowledgements.- 7. References.- The Application of Structural Modeling to Metabolic Mapping of Functional Neural Systems.- 1. Introduction.- 2. General Method.- 2.1. Building a Model.- 2.2. Preparation of Data.- 2.3. Running the Analysis.- 2.4. Stacked Models.- 2.5. Theoretical Interpretation of the Model.- 3. Auditory System Model of Long-Term Habituation.- 3.1. Results.- 3.2. Discussion of Auditory System Models.- 4. Visual System Model of the Effects of Patterned Light and Footshock.- 4.1. Results.- 4.2. Discussion of Visual System Models.- 5. General Discussion.- 5.1. Considerations and Caveats.- 5.2. Other Methods of Data Quantification.- 5.3. Concluding Remarks.- 6. References.- II: Application.- A 2-DG Analysis of the Effects of Monocular Deprivation on the Rat Visual System.- 1. Introduction.- 2. General Methods.- 2.1. Subjects.- 2.2. 2-DG Procedure.- 3. Results and Discussion.- 3.1. Six-Week Infant MD; Test: Binocular Exposure to Gratings.- 3.2. Six-Week Infant MD; Test: Dark Box.- 3.3. Six-Week Juvenile and Adult MD; Test: Binocular Exposure to Gratings.- 3.4. Six-Week Infant MD; Test: Binocular Exposure to Diffuse Light.- 3.5. Two- to Nine-Week Normal; Test: Monocular Exposure to Gratings or Flashing-Diffuse Light.- 4. Conclusions.- 5. Acknowledgements.- 6. References.- Development of Sound Representation in the Auditory Cortex of Tree Shrews (Tupaia Belangeri): A [14C]-2-DG Study.- 1. Introduction.- 2. Material and Methods.- 2.1. Subjects and Housing.- 2.2. 2-DG Application.- 2.3. Acoustic Stimulation.- 2.4. Tissue Preparation and Autoradiography.- 2.5. Histological Identification of Labelled Regions.- 2.6. Data Analysis and Documentation of Results.- 3. Results.- 3.1. Effect of Different Sound Patterns (BBN, HFS, LFS, SC) on 2-DG Uptake in Auditory Cortex of Adult Tree Shrews.- 3.2. Effect of Different Sound Patterns (BBN, SC) on 2-DG Uptake in the Auditory Cortex of Developing Tree Shrews.- 4. Conclusions.- 5. References.- Integration of Circadian and Visual Function in Mammals and Birds: Brain Imaging and the Role of Melatonin in Biological Clock Regulation.- 1. Introduction.- 1.1. The Avian Circadian Clock is a Multi-Component System.- 1.2. Mammalian Circadian Rhythmicity and the Suprachiasmatic Nucleus.- 2. Methodology.- 2.1. Circadian Rhythm Research.- 2.2. 2-Deoxyglucose Technique.- 2.3. In Vitro 2-[125 I] Iodomelatonin Binding.- 2.3. Image Representation.- 3. Melatonin and the Rat Circadian System.- 3.1. Effects of Melatonin on Rat SCN 2-Deoxyglucose Uptake.- 3.2. 2-[125I] Iodomelatonin Binding in the Rodent Brain.- 4. Sites of Melatonin Action in the Avian Brain.- 4.1. Circadian Variation of 2-Deoxyglucose Uptake in the Brain of the House Sparrow.- 4.2. Effect of Exogenous Melatonin on Cerebral 2-Deoxyglucose Uptake in the House Sparrow.- 4.3. 2-[125I]ldomelatonin Binding within the Avian Brain.- 5. Conclusions.- 6. Literature Cited.- Functional Correlates of Acute Prolonged Pain in the Rat Central Nervous System: 2-DG Studies.- 1. Introduction.- 2. Methods.- 2.1. The Formalin Test.- 2.2. Quantitative 2-DG Studies.- 2.3. Semi-Quantitative 2-DG Studies.- 3. Results and Discussion.- 3.1. Animal Behavior.- 3.2. Physiological Variables.- 3.3. Glucose Metabolism.- 4. Conclusions.- 5. Acknowledgements.- 6. References.- Brain Systems Involved in Depressed Behaviors: Corroboration from Different Metabolic Mapping Studies..- 1. Introduction.- 2. Methods.- 3. Results.- 4. Mapping Studies of Reward and Arousal.- 5. Mapping Studies of Reduced Locomotion.- 6. Conclusions.- 7. References.- 2-DG and Neuroethology: Metabolic Mapping of Brain Activity During Species-Typical Sexual and Aggressive Behaviors.- 1. Introduction.- 2. Reptiles as a Source of Model Systems in Behavioral Neuroscience.- 3. Brain Imaging During Sexual and Agonistic Behavior in Three Reptile Animal Models.- 3.1. Red-Sided Garter Snake.- 3.2. Green Anole Lizard.- 3.3. Whiptail Lizard.- 4. Conclusions.- 5. Acknowledgements.- 6. References.- Investigations into Time-Dependent Metabolic Changes During Memory Processing in the Mouse Brain Using (14C)-Deoxyglucose and (14C)-Glucose.- 1. Introduction.- 1.1. The Concept of Memory Processing.- 1.2. The Use of 2-DG to Study Learning and Memory Processes.- 2. Materials and Methods.- 2.1. Subjects.- 2.2. Surgery.- 2.3. Procedure for Injection of the Tracer.- 2.4. Processing of Brain Tissue.- 2.5. Densitometric Analysis of Autoradiographs: The Semi-Quantitative Method.- 2.6. The use of (14C)-Glucose to Study Short-Time Frames.- 2.7. Behavioral Tasks.- 2.8. Experimental Protocols.- 3. Results.- 3.1. Time-Dependent Sequential Increases in 2-DG Uptake During Memory Consolidation of the Bar-Pressing Task.- 3.2. Time-Dependent (14C)-Glucose Uptake Patterns During Memory Processing of Spatial Discrimination Testing in Radial Maze..- 3.3. General Discussion.- 4. Conclusions.- 5. Acknowledgements.- 6. References.- Localization of Learning-Related Metabolic Changes in Brain Structures of Common Toads: A 2-DG-Study.- 1. Introduction.- 2. Methods.- 2.1. Training Procedure.- 2.2. Mapping of Brain Activity with 2-DG, Technical Questions.- 3. Results.- 3.1. Behavioral Effects of Visual Conditioning on Prey-Predator Discrimination.- 3.2. Comparison of 2-DG Uptake on Naive and Visually Monocular and Binocular Conditioned Toads.- 3.3. Brain Lesions.- 3.4. Summary of Olfactory Learning Experiments in Toads with 2-DG.- 3.5. Summary of Extracellular Single Cell Recording.- 3.6. Summary of Visual Long-Term Habituation Experiments with 2-DG in Toads.- 3.7. Summary of Results of 2-DG Experiments in MS222 Anesthetized Toads.- 3.8. Summary of Results of 2-DG Experiments in Electrically RET Stimulated Toads.- 3.9. Summary of Results of 2-DG Experiments, Combining MS222 Anesthesia and Electrical RET Stimulation in Toads.- 4. Discussion.- 4.1. Associative Conditioning versus Extinction Learning.- 4.2. Usefulness of the Selected 2-DG Method in Combination with other Techniques for the Questions, Tackled in the Described Experiments..- 4.3. Physical Properties of the Stimulus and its Learned Motivational Significance.- 4.4. Changes of Metabolic Activity in the Central Visual Pathway.- 4.5. Changes of 2-DG Uptake Patterns in Non-Visual Forebrain Areas of Trained Toads.- 5. Conclusions.- 6. References.- Learning-Related Plasticity of Gerbil Auditory Cortex: Feature Maps versus Meaning Maps.- 1. Introduction.- 2. Basic Functional Organization of Auditory Cortex: Electrophysiology.- 3. Basic Functional Organization of Auditory Cotrex: Deoxyglucose Labeling.- 4. Learning-Induced Changes of Fluoro-2-Deoxyglucose Uptake.- 5. Electrophysiological Changes after Classical Conditioning.- 6. Discussion.- 7. Literature.- III: Discussions.- Discussions on Advances in Metabolic Mapping Techniques.- Discussions on Brain Imaging of Behavioral Functions.- Discussions on Brain Imaging of Learning Functions.