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.Role of Neurogenomics in the Development of Personalized Neurology
Personalized medicine is defined by the author as an integration of various biotechnologies to provide treatment best suited for a patient taking into consideration genomic as well as non-genomic factors. This chapter focuses on the role of neurogenomics in the development of personalized neurology, and the relationship to other relevant “omics” such as neuroproteomics is pointed out. Several of the genes relevant to neurological disorders have been sequenced, and their relation to various neurological disorders is under investigations. Genomic technologies may play a role in molecular diagnostics, biomarkers, ...
Source: Springer protocols feed by Neuroscience - January 1, 2015 Category: Neuroscience Source Type: news
Single-Cell Electroporation for In Vivo Imaging of Neuronal Morphology and Growth Dynamics
Single-cell electroporation (SCE) is a technique for acutely transfecting or dye-labeling individual neurons within intact living tissues. In addition to fluorescently labeling neurons, SCE can be used to conduct cell-autonomous studies of protein function by co-delivering fluorophores with DNA, RNA, antisense constructs, peptides, proteins, or drugs. SCE involves inserting a thin glass pipette into neural tissue to restrict an electric field and exposure to a solution of delivery compounds to an individual neuron at the pipette tip. Application of a brief electric pulse induces transient pores in the target cell and ionto...
Source: Springer protocols feed by Neuroscience - November 10, 2014 Category: Neuroscience Source Type: news
Simultaneous Collection of In Vivo Functional and Anatomical Data from Individual Neurons in Awake Mice
Ideally, to trace neural circuits, one often desires access to functional data that may be linked to anatomical attributes such as neuron type or projection target. Here we describe methods used for this purpose in our laboratory. We aim for this chapter to serve as a practical guide to applying “loose patch” or “cell-attached patch” electrophysiology techniques in vivo to simultaneously obtain information about neuronal firing (e.g., responses to sensory stimuli) and detailed anatomical information including dendritic morphology and axonal targeting. Since data on neuronal circuit function are ofte...
Source: Springer protocols feed by Neuroscience - November 10, 2014 Category: Neuroscience Source Type: news
Retrograde Tract-Tracing “Plus”: Adding Extra Value to Retrogradely Traced Neurons
Classical neuroanatomical tract-tracing methods have formed the basis for most of our current understanding of brain circuits. However, to obtain a deeper knowledge of the main operational principles of the brain, the simple delineation of brain connectivity is not sufficient. This particularly holds true in regard to the analysis of connections within the diseased brain, for instance, the study of a number of major neurological disorders through the use of animal models. In other words, the information gathered from tract-tracing techniques is often too static, and recent findings in the fields of neurophysiology, recepto...
Source: Springer protocols feed by Neuroscience - November 10, 2014 Category: Neuroscience Source Type: news
Wheat Germ Agglutinin (WGA) Tracing: A Classic Approach for Unraveling Neural Circuitry
Neuroanatomical tracing is a fundamental technique that has long been considered the primary method for visualizing brain networks in all areas of neuroscience. Although there are many new approaches for tracing neuronal connections, the lectin-based wheat germ agglutinin (WGA) tracing approach is still widely used, and it is firmly regarded as a classic method in the field. WGA has been used extensively to unravel both simple and complex neural networks in the central and peripheral nervous systems. It is reliable and versatile, as projections are labeled in the anterograde and retrograde directions. It is robust enough f...
Source: Springer protocols feed by Neuroscience - November 10, 2014 Category: Neuroscience Source Type: news
A Survey of Current Neuroanatomical Tracing Techniques
This chapter provides a systematic description of neuroanatomical tracing methods, with a brush of history. Tracing can be based on uptake and transport of tracer in living neurons but can also be based on physical diffusion in living neurons after intracellular injection of tracer or, in fixed tissue as is the case of Golgi silver staining, based on complex anorganic chemical reactions. Because of the special fixation status of human brain tissue, the physicochemical methods are prominent with this kind of nervous tissue. Nowadays, the transport methods enjoy popularity in animal connectivity models because they produce f...
Source: Springer protocols feed by Neuroscience - November 10, 2014 Category: Neuroscience Source Type: news
Genetic Pathways to Circuit Understanding in Drosophila
Genetic tools enable a diverse array of experimental approaches to dissect the relationships between brain function and behavior. Many genetic tools have been developed for use in the fruit fly, Drosophila melanogaster, making it a powerful model system due to its rich behavioral repertoire on one hand and, on the other, the accessible size and scale of its highly stereotyped nervous system. The stereotypy of the fly brain, in particular, makes it possible to interrogate, in essence, the same cell types—genetically, morphologically, physiologically, molecularly, and behaviorally—across individual flies. In this...
Source: Springer protocols feed by Neuroscience - November 10, 2014 Category: Neuroscience Source Type: news
Genetic Labeling of Synapses
A major challenge in neuroscience is to unravel how the synaptic contacts between neurons give rise to brain circuits. A number of techniques have been developed to visualize the synaptic organization of neurons. In this chapter, we focus on genetic methods to mark specific types of synapses so that synaptic sites can be visualized throughout the entire dendritic or axonal arbor of single neurons. Genetic synaptic labeling can be achieved by cell-type-specific viral or transgenic delivery of synaptic proteins tagged by fluorescent proteins. Sparse genetic labeling of neurons permits semiautomated quantification of the dist...
Source: Springer protocols feed by Neuroscience - November 10, 2014 Category: Neuroscience Source Type: news
Engineered Rabies Virus for Transsynaptic Circuit Tracing
Transsynaptic tracing using modified rabies virus (RV) is a powerful new technology in neuroscience that allows for visualization of targeted neurons and their synaptic connections. Here, we describe how a genetically engineered version of RV can be used for transsynaptic tracing studies of mammalian neuronal cells by providing protocols for viral isolation, propagation, pseudotyping, and concentration. The resulting genetically modified RV shows neuronal infectivity both in vitro and in vivo. Once the target neuron has been infected, the RV replicates and “jumps” presynaptically to connected neurons to provide...
Source: Springer protocols feed by Neuroscience - November 10, 2014 Category: Neuroscience Source Type: news
Hydrolytically Degradable Polyethylene Glycol (PEG) Hydrogel: Synthesis, Gel Formation, and Characterization
Degradable tight-mesh hydrogel scaffolds are a promising cell carrier for neural transplantation because they can match the stiffness of the native tissue, decrease immunogenicity of the transplant, and degrade over time as the new tissue forms. Here we describe the synthesis and gel formation of a hydrolytically degradable polyethylene glycol (PEG) hydrogel, where the mild gelation conditions allow for cell encapsulation prior to gel formation. We also describe how to measure swelling ratio and mass loss of the hydrogel over time in order to assess hydrogel degradation. (Source: Springer protocols feed by Neuroscience)
Source: Springer protocols feed by Neuroscience - November 4, 2014 Category: Neuroscience Source Type: news
Engineered Microdevices to Study and Manipulate Neural Stem Cell Chemotaxis
Traditional in vitro chemotaxis tools are typically limited by unstable concentration gradients, presence of hydrodynamic shear, and the inability to decouple cell migration directionality and speed. These limitations have restricted the reproducible and quantitative analysis of neuronal migration, which is a requirement for mechanism-based studies that may guide the development of new therapeutic strategies for neural regeneration. Here we describe a microfluidic setup for creating stable, linear, and shear-free gradients of putative chemotactic cues and their application in quantifying chemotaxis (i.e., migration directi...
Source: Springer protocols feed by Neuroscience - November 4, 2014 Category: Neuroscience Source Type: news
Cultivation and Imaging of Astrocytes on Protein-Coated Fluorescent Topographies Constructed from Aligned PLLA Electrospun Fibers
Electrospun poly-l-lactic acid (PLLA) fibers are presently explored as tissue engineering platforms for regeneration of the central nervous system. In particular, aligned, electrospun fibers are capable of directing astrocyte cellular extension and migration. The precise mechanisms by which aligned, electrospun substrates alter glial cell behavior are poorly understood. Therefore, there is a need for designing and refining electrospun fiber platforms and developing novel approaches for studying astrocytic behavior and physiology on aligned substrates. Here, we describe and discuss methods for (1) fabrication of fluorescent...
Source: Springer protocols feed by Neuroscience - November 4, 2014 Category: Neuroscience Source Type: news
Evaluating the Spatial and Temporal Protein Production in Neural Tissue Engineering Constructs In Vitro
Tissue engineering constructs are uniquely positioned not only for in vivo regenerative therapeutic applications but also to evaluate cellular responses to modifications in finely tuned three-dimensional (3-D) microenvironments as opposed to traditional two-dimensional (2-D) culture systems. However, moving into a 3-D system presents complications of assessing spatial and temporal alterations in cellular behavior (e.g., protein synthesis, motility, etc.). Here, we describe methods to evaluate spatial and temporal protein production in neural tissue engineering construct. (Source: Springer protocols feed by Neuroscience)
Source: Springer protocols feed by Neuroscience - November 4, 2014 Category: Neuroscience Source Type: news
Modulation of Neural Stem Cell Expressed Extracellular Matrix (ECM) by Targeting Glycosyltransferases
The stem cell niche microenvironment is rich in highly glycosylated extracellular matrix (ECM) proteins such as tenascin-C, chondroitin sulphate proteoglycans (CSPGs), laminins and their receptors, the β1-integrins. These proteins are present in the central nervous system (CNS) during embryonic development, and they are retained in adult neural stem cell niches such as the subventricular zone (SVZ) of the lateral ventricle. This niche composition favours stem cell maintenance, the proliferation of neuronal precursor cells and their differentiation. In this chapter we will briefly introduce three common glycan motifs p...
Source: Springer protocols feed by Neuroscience - November 4, 2014 Category: Neuroscience Source Type: news
Chondroitinase Gene Therapy for Spinal Cord Injury
Spinal cord injury (SCI) can lead to permanent paralysis below the level of injury. Environmental factors within the injured spinal cord have been shown to strongly participate in regenerative failure. Shortly after injury, the formation of a glial-fibroblastic scar develops at the injury site. Besides isolating the injury area and stabilizing inflammation and cellular damage, the glial scar is also an important source of both the physical and molecular barriers affecting axonal regeneration. The responding reactive astrocytes are known to secrete a group of potent axonal growth-inhibitory matrix molecules known as chondro...
Source: Springer protocols feed by Neuroscience - November 4, 2014 Category: Neuroscience Source Type: news
