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Electrophysiological and Biochemical Studies of AMPA Receptor Phosphorylation and Synaptic Plasticity in Hippocampal CA1 Mini-Slices

In the hippocampus, synapses undergo activity-dependent changes in synaptic strength, and this is thought to underlie some forms of learning and memory. Different patterns of activity can selectively enhance synaptic transmission, inducing long-term potentiation (LTP), or wea ...

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The Sagittally Sectioned Rat Hindbrain Preparation: Improved Access to the Brainstem Respiratory Network

Neurons that control breathing are distributed along the ventral respiratory column (VRC) in the ventrolateral brain stem, extending from the pons to the spinomedullary junction. These networks can be isolated in the en bloc brainstem-spinal cord preparation, which remains sponta ...

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Converging Methodologies in a Mammalian Simple System Focused on the Biology of Memory: Conditioned Odor Preference in the Neonate Rat

We present here a critical period model for odor preference learning in the rat pup, which can produce short-term or lifelong changes in odor-related behaviors depending on training parameters. Features which make it a powerful tool for the analysis of memory processes are described. The cha ...

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Electrophysiological Methods for Investigating Inhibitory Synaptic Plasticity

This chapter provides a detailed electrophysiological protocol for investigating inhibitory synaptic plasticity using the gramicidin perforated patch clamp technique. Gramicidin is a polypeptide antibiotic that is used as a perforating agent because it forms pores in the ne ...

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Laminar Quantification of Dendrites in Dentate Gyrus Granule Neurons

Synaptic integration of young neurons requires the sprouting and extension of dendrites in order to establish contacts with afferent neurons. For this reason, dendritic growth is often used in developmental studies as an important indicator of neuronal health and maturity. In the hipp ...

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Glutamate Receptors and Synaptic Physiology in Developing Neural Circuits

The major excitatory neurotransmitter in the CNS is glutamate. It binds to and activates fast, ligand-gated receptors such as AMPA and NMDA ionotropic receptors. AMPA receptors are composed of 4 subunits (GluA1–4) with varying stoichiometries. The subunit composition of the AMPA recep ...

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Translational Regulation of Synaptic Plasticity

Synaptic plasticity defines the process by which synapses, the connections between neurons, can be modified in response to activity. Plasticity can be either positive or negative, with strengthening and weakening of synapses occurring with distinct patterns of activity (neuronal ...

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Exploring Mechanisms of Synaptic Plasticity Using Exogenous Expression of Proteins at the Sensory-to-Motor Neuron Synapse of Aplysia

The use of expression constructs to drive exogenous expression of proteins has long been a pillar of cell and molecular biology. In this chapter, we will focus on two particular uses for this technique in studying synaptic plasticity, using Aplysia californica as a model: first, the use of overexp ...

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Sindbis Virus-Mediated In Vivo Expression of Recombinant CREB to Study Gene Function in Neuronal Plasticity and Behavior

Understanding the role of specific genes in diverse brain functions is a major challenge in neuroscience. To address this issue, we have optimized an in vivo transgene expression system based on the sindbis virus. In this chapter, we provide a detailed description of the cloning, production, and ...

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Conditional Gene Targeting: A Refined Method for Genetic Studies in Neuroscience

Conditional mutagenesis in mice is a key approach in Neuroscience that makes it possible to investigate the functions of defined genes within certain neural subpopulations. The approach is based on the combination of transgenic expression of a recombinase with targeted candidate ge ...

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Astrocytes

The central nervous system (CNS) is characterized by an immense structural complexity with intimate associations of its constituent cell types: neurons and different types of glial cells, mainly oligodendrocytes and astrocytes. A century ago (1893) described what are now called pro ...

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Neurons

There are several preparations that can be utilized to study the development of the vertebrate nervous system and the physiology and pharmacology of individual vertebrate neurons. These include both in vivo and in vitro preparations. The latter include brain slices (both conventional ...

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Organotypic Slice Explant Roller-Tube Cultures

Within the past decade, organ cultures have reemerged as an important complement to in vivo studies for examining regulatory mechanisms in various neuronal systems. The principle behind organ cultures is to maintain a tissue explant in a state as close as possible to that found in vivo, i.e., to pres ...

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Three-Dimensional Organ Culture Systems

Normal brain functions are to the highest degree dependent on the cytoarchitecture and the intercellular relationships that govern both the nervous system metabolism (for instance, the integrity of the blood-brain barrier) and specific functions (synaptic transmission, glio ...

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The Role of Cell Adhesion Molecules in Neurite Growth

Stereotyped patterns of axonal growth and axon-axon bundling leading to tract formation are common features of nervous system development in organisms as diverse as grasshoppers and humans. In grasshoppers, axons growing from individual, identified neurons follow stereotyped ...

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Detection and Analysis of Growth Factors Affecting Neural Cells

Individual cellular behaviors are controlled by extrinsic signals within the environmental milieu. These signals maybe ions, nutrients, hormones, or, pertinent to this review, a collection of proteins generally termed “growth factors.” Growth factors, which may have autocrine, p ...

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Environmental Influences on Cells in Culture

The technique of culturing cells derived from the nervous system has been in use for over 80 years, with the objective of simplifying the experimental system to provide readily manipulatable models of neural function. These preparations are useful for testing hypotheses relevant to cell a ...

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Identification of Cell Types in Neural Cultures

Tissue culture provides an opportunity to study the functions of the nervous system under strictly controlled conditions. As amply documented in other chapters in this volume, the culture medium, the surface upon which the cultures are growing, and various cellular and soluble factors can ...

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Basic Techniques for Cell Culturing

Until recently, the cell culture technique was employed as the sole biological system within a laboratory, and major efforts were concentrated on the setting up and running of such a cell culture laboratory. However, recent developments in several areas, use on monoclonal antibodies and ce ...

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Brain Capillaries: Cell Culture of Capillary Endothelium Derived from Cerebral Microvessels

The microvasculature of the brain is important in the normal development and maintenance of the blood-brain barrier. Since glial cells are in close contact with brain capillaries in situ, communication between glial cells and capillary endothelial cells is thought to be important in mai ...

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