Springer protocols feed by Neuroscience 
This is an RSS file. You can use it to subscribe to this data in your favourite RSS reader or to display this data on your own website or blog.
This is an RSS file. You can use it to subscribe to this data in your favourite RSS reader or to display this data on your own website or blog.Characterization and Analysis of Extracellular Matrix in Malignant Brain Tumors and Their Cellular Derivatives
The neural extracellular matrix (ECM) is a key regulator of cellular phenotype in normal and diseased states. Analytical approaches have helped uncover the myriad of roles the ECM plays in a variety of diseases including malignant gliomas, the most prevalent and deadly primary brain tumors. Major components of the glioma ECM include secreted proteoglycans as well as basal lamina proteins such as laminins. Characterization of secreted proteoglycans by protein blotting is an invaluable tool to understand their molecular complexity. To provide complete characterization of these molecules, tissue specimens must be first proces...
Source: Springer protocols feed by Neuroscience - November 4, 2014 Category: Neuroscience Source Type: news
Culturing Neurons, Glia, and Progenitor Cells in Three-Dimensional Hydrogels
The physiologic environment of neuronal and glial cells is the three-dimensional (3D) tissue. We and others have demonstrated that more physiologically relevant cell responses occur in 3D culture environments as opposed to flat stiff culture substrates such as tissue culture plates and coverslips. Hydrogels provide significant advantages towards providing cells a biocompatible, all-encompassing culture environment, but pose challenges towards adapting standard biochemical and molecular biology assay techniques. Herein, we provide methods for encapsulating and culturing cells inside 3D type I collagen gels. We also provide ...
Source: Springer protocols feed by Neuroscience - November 4, 2014 Category: Neuroscience Source Type: news
Replicating the In Vivo Environment: Organotypic and Submerged Three-Dimensional Culture Methods
In the embryo, soluble molecules are expressed in gradients to regulate tissue morphogenesis and direct cell lineage. Yet most tissue culture methods rely on modified two-dimensional (2D) substrates. When a cell is attached to a flat surface, the cellular structure is limited by adhesion to the substrate, and the cell is unable to acquire the in vivo morphology and possibly function. In most tissue culture applications, extracellular matrix (ECM) molecules are presented on the basal planar 2D substrate. In addition, cell-secreted ECM deposits on the 2D substrate or is released into the liquid media. Establishing concentrat...
Source: Springer protocols feed by Neuroscience - November 4, 2014 Category: Neuroscience Source Type: news
Extracellular Matrix Components as a Substrate for Outgrowing Motoneurons
Embryonic motoneurons depend on extracellular matrix (ECM), and molecules within which are potent mediators of survival, axonal growth, and guidance, exhibiting either attractive or repellent functions. Therefore, isolated embryonic spinal cord motoneurons were cultured on a three-dimensional substrate composed of different ECM molecules produced by different glia cell lines. The motoneuron-matrix co-culture could be used to analyze the modifying effects of matrix components like chondroitin sulfate proteoglycans, bifunctional molecules with modifying properties either due to their protein backbone or the glycan portion su...
Source: Springer protocols feed by Neuroscience - November 4, 2014 Category: Neuroscience Source Type: news
High-Throughput Quantitative Assay for Analyzing Neurite Outgrowth on a Uniform Substratum: The Cell-Substratum Assay
Neurite outgrowth measurements often require time-consuming measurements of multiple images. The numerous experimental conditions lead to long analysis time and effort. A high-throughput assay provides a quick method to obtain quantitative measures of neurite outgrowth. In this chapter, the experimental setup and imaging protocol are described to efficiently process images of primary neurons or cell lines in order to quantitate parameters. (Source: Springer protocols feed by Neuroscience)
Source: Springer protocols feed by Neuroscience - November 4, 2014 Category: Neuroscience Source Type: news
Techniques for Examining the Effect of Substratum-Bound Proteoglycans on Neurite Outgrowth In Vitro
Techniques to investigate mechanisms affecting neurite outgrowth, in vitro, are invaluable to the neuroscience community. In particular, these techniques have enhanced our understanding of the role of the extracellular matrix (ECM) in neuronal regeneration following spinal cord injury (SCI). In this chapter, we explain two useful techniques that have been used in our laboratory to investigate the role of chondroitin sulfate proteoglycans (CSPGs) in the inhibition of neurite outgrowth: (1) the Stripe Assay (Snow et al., Exp Neurol 109(1):111–130, 1990), which is a widespread model used to determine the effect of a pat...
Source: Springer protocols feed by Neuroscience - November 4, 2014 Category: Neuroscience Source Type: news
Isolation of Perinatal Mouse Cerebral Cortical Astrocytes
Proper functioning of the central nervous system requires collaboration between neurons and glia. Glia are composed of astrocytes, oligodendrocytes, and microglia. In vitro, astrocytes provide support for neuronal cultures for the study of development and regeneration of the brain. Upon injury, astrocytes can produce molecules that protect the harmed area and also prevent further regeneration. Research of astrocyte cell biology and translational uses requires isolation of purified astrocytes. The classical methods for preparing rat astrocytes have been adapted for mouse. In this chapter, we outline a simple method for prep...
Source: Springer protocols feed by Neuroscience - November 4, 2014 Category: Neuroscience Source Type: news
Investigating Cell-ECM Interactions and ECM Synthesis in Three-Dimensional Hydrogels
Model systems composed of cells cultured within three-dimensional (3D) gels are widely used to examine mechanisms of morphogenesis and repair and are applied as vehicles for delivering cells for therapeutic applications in vivo. Cells secrete extracellular matrix (ECM), the composition of which depends on the type of cell and cues present; the ECM is assembled and remodeled, forming a dynamic cellular niche. While it is generally recognized that the presence of ECM and cellular interactions with the ECM, for example via integrin binding, are closely tied to cell and tissue behavior, few investigations have centered on quan...
Source: Springer protocols feed by Neuroscience - November 4, 2014 Category: Neuroscience Source Type: news
Perineuronal Nets: A Special Structure in the Central Nervous System Extracellular Matrix
The extracellular matrix of the central nervous system (CNS) contains various compartments. Perineuronal nets (PNNs) are specialized and stable brain matrices which enwrap the surface of the soma and proximal dendrites on the neuronal surface. The presence of PNNs marks the end of the critical period in the developing CNS and limits the degree of plasticity after CNS injury. Chondroitin sulfate proteoglycans, link proteins, hyaluronan, and tenascin-R are identified in the PNNs. Here, we describe a method which allows the sequential extraction of the main matrix compartments of the CNS, ending with the isolation of the PNN ...
Source: Springer protocols feed by Neuroscience - November 4, 2014 Category: Neuroscience Source Type: news
The Use of Synaptic Basal Lamina and Its Components to Identify Sites of Recent Morphological Alterations at Mammalian Neuromuscular Junctions
While muscle fibers are ensheathed in basement membrane material, the extracellular matrix at their synaptic segment, the neuromuscular junction (NMJ), is specialized and contains factors that are essential for the proper development, maintenance, and regeneration of the synapse after injury. Immunohistochemical studies have been critical in characterizing the composition of the synaptic basal lamina at the NMJ. Although NMJs are extremely stable synapses, they can undergo structural alteration both pre- and postsynaptically, especially during development and in response to injury. Even though there are no permanent molecu...
Source: Springer protocols feed by Neuroscience - November 4, 2014 Category: Neuroscience Source Type: news
Immunohistochemical Characterization of Brain Neural and Vascular Basement Membranes
Basement membranes are a critical component of the neural and vascular structures of the brain and play important roles in both brain development and function. Defects in basement membrane integrity are associated with a large number of brain disorders including cobblestone lissencephaly and intracerebral hemorrhage. Here, we describe immunohistochemical approaches for characterizing the neural and vascular basement membranes in the perinatal mouse brain. (Source: Springer protocols feed by Neuroscience)
Source: Springer protocols feed by Neuroscience - November 4, 2014 Category: Neuroscience Source Type: news
Speech Mapping with Transcranial Magnetic Stimulation
The identification of eloquent cortical regions associated with language function is of great relevance to basic scientists as well as clinicians. Repetitive transcranial magnetic stimulation (rTMS) is a relatively novel technique that allows for noninvasive, lesion-based identification of critical language sites. The integration of frameless stereotactic navigation systems with rTMS allows for precise targeting and identification of the cortical region that is being stimulated. Integration of this navigational data also allows for accurate, real-time analysis of magnetic field strength in the stimulated region of interest...
Source: Springer protocols feed by Neuroscience - May 22, 2014 Category: Neuroscience Source Type: news
Navigated Transcranial Magnetic Stimulation: Principles and Protocol for Mapping the Motor Cortex
Transcranial magnetic stimulation (TMS) is a unique method for studying the human brain. Whereas the majority of imaging tools detect and map all the brain areas that participate during a given task (both primary and secondary network activations), TMS, when used to evoke a measurable physiological response, maps only those areas that are mandatory for the observed reaction. As such, TMS is particularly suitable for mapping cortical motor areas and for assessing the functional status of the motor tracts, both in normal subjects and in patients. In this chapter, we explore the physical and mechanistic background of using TM...
Source: Springer protocols feed by Neuroscience - May 22, 2014 Category: Neuroscience Source Type: news
Protocol for Depression Treatment Utilizing H-Coil Deep Brain Stimulation
The Brainsway Deep TMS System for the treatment of major depressive disorder (MDD) was cleared by the FDA on January 2013 for the treatment of depressive episodes in adult patients suffering from MDD who failed to achieve satisfactory improvement from previous antidepressant medication treatment in the current episode, based on a large, prospective, double blind, randomized, controlled multicenter trial. This chapter presents the detailed protocol of this multicenter trial. (Source: Springer protocols feed by Neuroscience)
Source: Springer protocols feed by Neuroscience - May 22, 2014 Category: Neuroscience Source Type: news
A Review of Current Clinical Practice in the Treatment of Major Depression
Transcranial magnetic stimulation (TMS) is a method of using high-strength pulsed magnetic fields to induce a depolarizing electric current in a localized area of the cerebral cortex. A large, replicated body of multisite, randomized sham-controlled studies has established the safety and effectiveness of a specific TMS device for the treatment of patients with pharmacoresistant major depressive disorder. This chapter reviews the current understanding of the physiologic effects and presumed mechanism of effect of TMS as an antidepressant. In addition, the scientific evidence base for its safety and efficacy are reviewed, al...
Source: Springer protocols feed by Neuroscience - May 22, 2014 Category: Neuroscience Source Type: news
