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Short Protocols in Neuroscience: Systems and Behavioral Methods


Description: As recently as 25 years ago, a wide technical gulf separated neuroscience from the rest of biology. To address the complex anatomy and unusual physiological properties of neurons, neurobiologists (a term not then in vogue) used highly specialized techniques of anatomical tracing and electrophysiological analysis that were foreign to most biologists. Conversely, many neurobiologists, absorbed in their own disciplines, were insulated from the ideas and techniques in common use elsewhere in biology. Twenty-five years later the technical landscape of neurobiology is vastly different. The specialized features of neurons are now recognized as variations on fundamental biological processes shared by all cells. Axonal transport and neurotransmitter release, for example, are now known to occur by mechanisms that are related to those of membrane trafficking and secretion used by all eukaryotic cells, and the neurofilaments whose unusual staining properties allowed neuroanatomists to trace axonal tracts are now recognized as part of a larger family of intermediate filaments found in many cell types. The recognition of common themes in the biology of neurons and other cells has opened neuroscience to the wide range of techniques used in the study of other cell types. Of these, molecular biological approaches, in particular, have had an enormous impact on neurobiology. Immunological, genetic, cell culture, and cell imaging techniques have also enriched the repertoire of cellular neurobiologists. In the meantime, innovations in biophysics have allowed the electrical properties of cells to be studied at the level of single channels. Methods for preserving brain microcircuitry through the use of brain slices have allowed the complex organization of the mammalian brain to be studied at the cellular level. Finally, the techniques used by neurobiologists have been amplified by our increasing ability to study the biological bases of behavior. These exciting developments have posed a considerable challenge to neurobiologists in that the array of relevant technologies for study of the nervous system is arguably broader than for biologists in any other field. In a single experiment, laboratories may attempt to relate results obtained with molecular, cellular, anatomical, and behavioral techniques. The demand for technical versatility is met in part through increased collaborations between laboratories and in part by broad training among neuroscientists. The contemporary pace of discovery and of technical innovation is such, however, that all of us are continually learning new techniques, either as practitioners or as knowledgeable collaborators.    


Contents: 1. NEUROPHYSIOLOGY (Michael A. Rogowski).
1.0 Introduction.
1.1 Fabrication of Patch Pipets.
1.2 Whole-Cell Voltage Clamp Recording.
1.3 Preparation of Hippocampal Brain Slices.
1.4 Synaptic Plasticity in Hippocampal slice Preparation.
1.5 Recording and Analyzing Synaptic Currents and Synaptic Potentials.
1.6 Voltage Clamp Recordings from Xenopus Oocytes.
1.7 Preparation and Maintenance of Organotypic Slice Cultures of CNS Tissues.
1.8 Patch Clamp Recording in Brain Slices.
1.9 Single-Channel Recording.
1.10 Acute Isolation of Neurons from the Mature Mammalian Central Nervous System.

1.11 Patch Clamp Recording from Neuronal Dendrites.

2. NEUROCHEMISTRY/NEUROPHARMACOLOGY (Phil Skolnick).
2.0 Introduction.
2.1 Scintillation Proximity Assay.
2.2 Methods for Sample Preparation for Direct Immunoassay Measurement of Analytes in Tissue Homogenates: ELISA.
2.3 Assessment of Cell Viability in Primary Neuronal Cultures.
2.4 Measurement of NO and NO Synthase.
2.5 Measurement of Oxygen Radical and Lipid Peroxidation in Neural Tissues.
2.6 Overview of Microdialysis.
2.7 Microdialysis in Rodents.
2.8 Microdialysis in Nonhuman Primates.
2.9 Detections and Quantification of Neurotransmitters in Dialysates.
2.10 Saturation Assays of Radioligand Binding to Receptors and Their Allosteric Modulatory Sites.
2.11 Uptake and Release of Neurotransmitters.
2.12 Measurement of Chloride Movement in Neuronal Preparations.
2.13 Measurement of Cation Movement in Primary Cultures Using Fluorescent Dyes.
2.14 Measurement of Second Messengers in Signal Transduction: cAMP and Inositol Phosphates.
2.15 In Vivo Measurement of Blood-Brain Barrier Permeability.

3. BEHAVIORAL NEUROSCIENCE (Michael A. Rogowski).
3.0. Introduction.
3.1. Assessment of Developmental Milestones in Rodents.
3.2. Locomotor Behavior.
3.3. Motor Coordination and Balance in Rodents.
3.4. Basic Measures of Food Intake.
3.5. Sexual and Reproductive Behavior.
3.6. Parental Behaviors in Rats and Mice.
3.7. Application of Experimental Stress in Laboratory Rodents.
3.8. Rodent Models of Depression: Forced Swimming and Tail Suspension Behavioral Despair Tests in Rats and Mice.
3.9. Rodent Models of Depression: Learned Helplessness Induced in Mice.
3.10. Animal Tests of Anxiety.
3.11. Assessment of Spatial Memory Using the Radial Arm Maze and Morris Water Maze.
3.12 Cued and Contextual Fear Conditioning in Mice.
3.13 Conditioned Flavor Aversions: Assessment of Drug-Induced Suppression of Food Intake.
3.14 Measurement of Startle Response, Prepulse Inhibition, and Habituation.
3.15 Latent Inhibition.

4. PRECLINICAL MODELS OF NEUROLOGIC AND PSYCHIATRIC BEHAVIORS (Phil Skolnick).
4.0 Introduction.
4.1 Preclinical Models of Parkinson’s Disease.
4.2 Rodent Models of Global Cerebral Ischemia.
4.3 Rodent Models of Focal Cerebral Ischemia.
4.4 Inducing Photochemical Cortical Lesions in Rat Brain.
4.5 Traumatic Brain Injury in the Rat Using the Fluid-Percussion Model.
4.6 Models of Neuropathic Pain in the Rat.
4.7 Dural Inflammation Models of Migraine Pain.
4.8 Animal Models of Painful Diabetic Neuropathy: The STZ Rat Model.
4.9 Models of Nociception: Hot Plate, Tail Flick, and Formalin Tests in Rodents.
4.10 Place Preference Tests in Rodents.
4.11 Intravenous Self-Administration of Ethanol in Mice.
4.12 Preclinical Models to Evaluate Potential Pharmacotherapeutic Agents in Treating Alcoholism and Studying the Ni….
4.13 Experimental Autoimmune Encephalomyelitis (EAE).
4.14 Models of Amyotrophic Lateral Sclerosis.
4.15 Measurement of Panic-Like Responses Following Intravenous Infusion of Sodium Lactate in Panic-Prone Rats.
4.16 Chemoconvulsant Model of Chronic Spontaneous Seizures.
Appendices.
References.
Index.




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