Springer protocols feed by Neuroscience 
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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.Alphavirus-Based Vectors
Alphavirus vectors have been developed for transient gene expression. The most common approach has been to generate replication-deficient vectors based on Semliki Forest virus, Sindbis virus, or Venezuelan equine encephalitis virus. All these alphavirus vectors have demonstrated efficient transduction of neurons in primary cultures, slices and in vivo. Wild-type vectors are cytotoxic resulting in host cell death. Mutant vectors have showed reduced toxicity and prolonged host cell survival. Engineering of temperature-sensitive mutants has allowed controlled expression and targeting to interneurons and astrocytes. Alphavirus...
Source: Springer protocols feed by Neuroscience - October 25, 2013 Category: Neuroscience Source Type: news
Herpes Simplex Virus 1 (HSV-1)-Based Vectors
Herpes simplex virus type 1 (HSV-1) is a neurotropic virus displaying several outstanding adaptations to the nervous system, and each of them can be rationally exploited in the design of gene therapy vectors with regard to neurological applications. Replication-competent attenuated vectors are becoming a suitable and powerful tool to eradicate brain tumors, such as malignant gliomas, due to their ability to replicate and spread only within the tumor mass, and have reached Phase II clinical trials in some cases. Replication-incompetent recombinant vectors are nontoxic gene transfer tools that preserve most of the neurotropi...
Source: Springer protocols feed by Neuroscience - October 25, 2013 Category: Neuroscience Source Type: news
Adeno-Associated Virus-Based Vectors
Viral vectors based on recombinant adeno-associated virus have gained increasing interest over the last two decades as promising delivery vehicles in gene therapy. This enthusiasm is based on their ability to infect a broad range of tissues including proliferative and quiescent cells, to establish long-term expression in vitro and in vivo, combine to an excellent safety profile, as they are replication deficient, poorly immunogenic, and have not been associated to any disease. This chapter provides detailed protocols for small-scale production and purification of adeno-associated vectors and currently used methods for the ...
Source: Springer protocols feed by Neuroscience - October 25, 2013 Category: Neuroscience Source Type: news
Recombinant Adenovirus in Neurobiology
Recombinant adenoviruses have been extensively applied in basic research and gene therapy. Recently, there is an increase in neurobiological studies using recombinant adenovirus technology to manipulate gene expression in neurological systems. Recombinant adenovirus technology provides a useful tool for investigating the function of a gene of interest, neurocircuitries and the delivery of genes for therapy. In order to take full advantage of this technique, it is important to understand the strategies for the design and application of the virus. The goal of this chapter is to provide a practical protocol on the application...
Source: Springer protocols feed by Neuroscience - October 25, 2013 Category: Neuroscience Source Type: news
Lentiviral Vectors as Research Tools in Neurobiology: Design and Production
Viral vectors are now common in contemporary neuroscience research and their use as gene transfer tools for the central nervous system has seen an enormous growth in the last 2 decades. This chapter discusses about designing, production, and use of lentiviral vectors (LVs), one of the most popular and versatile system currently available. (Source: Springer protocols feed by Neuroscience)
Source: Springer protocols feed by Neuroscience - October 25, 2013 Category: Neuroscience Source Type: news
The Cre/Lox System to Assess the Development of the Mouse Brain
Cre-mediated recombination has become a powerful tool to confine gene deletions (conditional knockouts) or overexpression of genes (conditional knockin/overexpression). By spatiotemporal restriction of genetic manipulations, major problems of classical knockouts such as embryonic lethality can be circumvented. Furthermore Cre-mediated recombination has broad applicability in the analysis of the cellular behavior of subpopulations and cell types as well as for genetic fate mapping. This chapter will give an overview about applications for the Cre/LoxP system and their execution. (Source: Springer protocols feed by Neuroscience)
Source: Springer protocols feed by Neuroscience - September 24, 2013 Category: Neuroscience Source Type: news
In Utero Electroporation to Study Mouse Brain Development
In utero electroporation is a rapid and powerful technique to study the development of many brain regions. This approach presents several advantages over other methods to study specific steps of brain development in vivo, from proliferation to synaptic integration. Here, we describe in detail the individual steps necessary to carry out the technique. We also highlight the variations that can be implemented to target different cerebral structures and to study specific steps of development. (Source: Springer protocols feed by Neuroscience)
Source: Springer protocols feed by Neuroscience - September 24, 2013 Category: Neuroscience Source Type: news
Immunohistochemistry and RNA In Situ Hybridization in Mouse Brain Development
During development, the mouse brain is progressively divided into functionally distinct compartments. Numerous neuronal and glial cell types are subsequently generated in response to various inductive signals. Each cell expresses a unique combination of genes encoding proteins from transcription factors to neurotransmitters that define its role in brain function. To understand these important and highly sophisticated processes, it is critical to accurately locate the various proteins and cells that produce them. In this chapter, we introduce the techniques of immunohistochemistry, which detects the localization of specific...
Source: Springer protocols feed by Neuroscience - September 24, 2013 Category: Neuroscience Source Type: news
RNAi-Based Gene Silencing in Chicken Brain Development
The mouse is the most commonly used vertebrate model for the analysis of gene function because of the well-established genetic tools that are available for loss-of-function studies. However, studies of gene function during development can be problematic in mammals. Many genes are active during different stages of development. Absence of gene function during early development may cause embryonic lethality and thus prevent analysis of later stages of development. To avoid these problems, precise temporal control of gene silencing is required. (Source: Springer protocols feed by Neuroscience)
Source: Springer protocols feed by Neuroscience - September 24, 2013 Category: Neuroscience Source Type: news
Transplantation of Neural Tissue: Quail-Chick Chimeras
Tissue transplantation is an important approach in developmental neurobiology to determine cell fate, to uncover inductive interactions required for tissue specification and patterning as well as to establish tissue competence and commitment. Avian species are among the favorite model systems for these approaches because of their accessibility and relatively large size. Here we describe two culture techniques used to generate quail-chick chimeras at different embryonic stages and methods to distinguish graft and donor tissue. (Source: Springer protocols feed by Neuroscience)
Source: Springer protocols feed by Neuroscience - September 24, 2013 Category: Neuroscience Source Type: news
Immunohistochemistry and In Situ Hybridization in the Developing Chicken Brain
One of the first steps in studies of gene function is the spatiotemporal analysis of patterns of gene expression. Indirect immunohistochemistry is a method that allows the detection of a protein of interest by incubating a histological section with an antibody or antiserum raised against the protein and then localizing this primary antibody with a tagged secondary antibody. To determine the cellular source of a protein of interest, or if a specific antibody is not available, specific transcripts can be localized using in situ hybridization. A histological section is incubated with a labeled RNA probe that is complementary ...
Source: Springer protocols feed by Neuroscience - September 24, 2013 Category: Neuroscience Source Type: news
Anatomical Dissection of Zebrafish Brain Development
Zebrafishbrain.org is an online neuroanatomical atlas of the embryonic zebrafish. The atlas uses high-resolution confocal images and movies of transgenic lines to describe different brain structures. This chapter covers detail of materials and protocols that we employ to generate data for the atlas. (Source: Springer protocols feed by Neuroscience)
Source: Springer protocols feed by Neuroscience - September 24, 2013 Category: Neuroscience Source Type: news
Dynamic Neuroanatomy at Subcellular Resolution in the Zebrafish
Genetic means to visualize and manipulate neuronal circuits in the intact animal have revolutionized neurobiology. “Dynamic neuroanatomy” defines a range of approaches aimed at quantifying the architecture or subcellular organization of neurons over time during their development, regeneration, or degeneration. A general feature of these approaches is their reliance on the optical isolation of defined neurons in toto by genetically expressing markers in one or few cells. Here we use the afferent neurons of the lateral line as an example to describe a simple method for the dynamic neuroanatomical study of axon te...
Source: Springer protocols feed by Neuroscience - September 24, 2013 Category: Neuroscience Source Type: news
Sensitive Whole-Mount Fluorescent In Situ Hybridization in Zebrafish Using Enhanced Tyramide Signal Amplification
We present here a sensitive fluorescent in situ hybridization method for colocalization analysis of different transcripts in whole embryonic zebrafish brains. The method is based on simultaneous hybridization of differently hapten-labeled RNA probes followed by sequential rounds of horseradish peroxidase (POD)-based transcript detection. Sequential detection involves enhancement of fluorescent signals by tyramide signal amplification (TSA) and effective inactivation of the antibody–POD conjugate prior to the following detection round. We provide a detailed description of embryo preparation, hybridization, antibody de...
Source: Springer protocols feed by Neuroscience - September 24, 2013 Category: Neuroscience Source Type: news
In Situ Hybridization and Immunostaining of Xenopus Brain
The dynamic expression pattern analysis provides the primary information of gene function. Differences of the RNA and/or protein location will provide valuable information for gene expression regulation. Generally, in situ hybridization (ISH) and immunohistochemistry (IHC) are two main techniques to visualize the locations of gene transcripts and protein products in situ, respectively. Here we describe the protocol for the whole brain dissection, the in situ hybridization and immunostaining of the developing Xenopus brain sections. Additionally, we point out the modification of in situ hybridization for microRNA expression...
Source: Springer protocols feed by Neuroscience - September 24, 2013 Category: Neuroscience Source Type: news
