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Culture of Rat Cerebellar Granule Neurons and Application to Identify Neuroprotective Agents

In primary culture of the early postnatal cerebellum, glutamatergic granule cells are highly enriched and recapitulate many properties characteristic of developing granule neurons in vivo. For example, withdrawal of K+ from differentiated rat primary cerebellar granule neur ...

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Rodent Spinal Cord Injury Model and Application of Neurotrophic Factors for Neuroprotection

Spinal cord injury (SCI) is a serious clinical problem that causes lifetime disabilities to victims and inflicting huge social burden on our society. One of the main lacunae in developing potential therapeutic measures in SCI is a lack of suitable animal models that could be comparable to clini ...

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Evaluating Motor Neuron Death in Neonatal Rats Subjected to Sciatic Nerve Lesion

Neonatal sciatic nerve lesion is a useful experimental model for the study of neuronal cell death. Sciatic nerve transection or crush is the most frequently used approach to evaluate motoneuron loss in the lumbar enlargement of the spinal cord. Here we describe and illustrate the surgical pro ...

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Brain Microdialysis in Freely Moving Animals

Brain microdialysis is an analytical technique used for the dynamic monitoring of brain neurochemistry in awake, freely moving animals. This technique requires the insertion of a small dialysis catheter, called a microdialysis probe, into a specific brain region, and its perfusion wi ...

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A 6-Hydroxydopamine In Vivo Model of Parkinsons Disease

Animal models of Parkinson’s disease are essential to explore pathophysiological hypotheses and to test new treatment options, including neurotrophic factors. Catecholaminergic neurotoxins used to generate such models are 6-hydroxydopamine and 1-methyl-4-phenyl-1,2 ...

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Neuronal Growth-Promoting and Inhibitory Cues in Neuroprotection and Neuroregeneration

During development of the nervous system, neurons extend axons over considerable distances in a highly stereospecific fashion in order to innervate their targets in an appropriate manner. This involves the recognition, by the axonal growth cone, of guidance cues that determine the path ...

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Rat Hippocampal Slice Culture Models for the Evaluation of Neuroprotective Agents

Organotypic slices cultured for weeks in vitro represent an extremely valuable strategy for the investigation of the long-term properties of neuronal circuits under physiological and pathological conditions. Here, we describe how to prepare rat organotypic hippocampal slice ...

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[3H]Serotonin Release Assay Using Antigen-Stimulated Rat Peritoneal Mast Cells

The concentration of nerve growth factor (NGF) is elevated in a number of inflammatory and autoimmune states in conjunction with increased accumulation of mast cells. Mast cells, which are of hematopoietic lineage, and NGF appear to be involved in neuroimmune interactions and tissue infl ...

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Preparation and Characterization of Biocompatible Chitosan Nanoparticles for Targeted Brain Delivery of Peptides

Here, we describe a nanocarrier system that can transfer chitosan nanoparticles loaded with either small peptides such as the caspase inhibitor Z-DEVD-FMK or a large peptide like basic fibroblast growth factor across the blood–brain barrier. The nanoparticles are selectively dire ...

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The Use of Specific AAV Serotypes to Stably Transduce Primary CNS Neuron Cultures

Although primary neuronal cell cultures are a valuable source of in vitro insight for many neurobiologists, all current gene expression technologies for these cells have significant drawbacks. Some of these limitations of current gene expression protocols include toxicity, tra ...

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Imaging Amyloid Precursor Protein In Vivo: An Axonal Transport Assay

Fusion of fluorescent probes to axonally transported proteins represents an established approach that enables live imaging of axonal transport. In this approach, in vivo examination of fluorescent particle dynamics provides information about the length, directionality, and ...

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High-Resolution Imaging and Evaluation of Spines in Organotypic Hippocampal Slice Cultures

Dendritic spines act as sites of excitatory neuronal input in many types of neurons. Spine shape correlates with the strength and maturity of synaptic contacts. Thus, evaluation of spine morphology is relevant for studies on neuronal development, for determination of morphological co ...

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Dissociated Cell Culture for Testing Effects of Carbon Nanotubes on Neuronal Growth

Cell culture has emerged as an important research method for studying the effects of carbon nanotubes (CNTs) on primary neurons. We describe the procedure for preparation of dissociated mixed cell culture from postnatal rat hippocampi. Based on morphological criteria and specific ne ...

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Neurite Outgrowth Assessment Using High Content Analysis Methodology

High content analysis of neurite outgrowth enables the rapid and comprehensive phenotypic assessment of individual neurons in a multiwell format amenable to high throughput assays. The resulting data are considered “high content” because multiple measurements of neuronal out ...

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Indirect Immunofluorescence Staining of Cultured Neural Cells

Immunofluorescence is a technique allowing the visualization of a specific protein or antigen in cells or tissue sections by binding a specific antibody chemically conjugated with a fluorescent dye such as fluorescein isothiocyanate. There are two major types of immunofluoresce ...

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Preparation and Culture of Adrenal Chromaffin Cells

Cultured chromaffin cells have been used for almost 40 years in the study of different cell functions using biochemical, electrophysiological, pharmacological, and toxicological approaches. Chromaffin cells are essentially secretory cells that are used to model sympathetic ...

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The Neurotrophin Family of Neurotrophic Factors: An Overview

The neurotrophins are a family of closely related proteins that were first identified as survival factors for sympathetic and sensory neurons and have since been shown to control a number of aspects of survival, development, and function of neurons in both the central and peripheral nervous s ...

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Compartmented Chambers for Studying Neurotrophic Factor Action

Neurotrophic factors released by target tissues maintain the survival and differentiation of innervating neurons. The manner by which these target-derived neurotrophic proteins communicate with innervating neurons has been actively pursued for over three decades. The pre ...

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Use of PC12 Cells and Rat Superior Cervical Ganglion Sympathetic Neurons as Models for Neuroprotective Assays Relevant to Parkinsons Disease

Cellular models composed of primary neuronal cultures or neuron-like cell lines are commonly used to study neuron cell death and to test the neuroprotective properties of specific compounds. Cellular models are easily accessible, permitting dissection and modulation of signali ...

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Culture and Proliferation of Highly Purified Adult Schwann Cells from Rat, Dog, and Man

This chapter presents fast and easy protocols to obtain highly purified cultures of proliferating adult rat, canine, and human Schwann cells. Cell preparation from predegenerated adult sciatic nerves combined with the use of melanocyte growth medium supplemented with forskolin, f ...

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