神经生物学

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 demonst ...

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 attenuat ...

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 cell ...

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 ...

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 popul ...

Motor neuron diseases such as amyotrophic lateral sclerosis (ALS) and spinal muscular atrophy (SMA) are characterized by the progressive loss of motor neurons in the spinal cord and primary motor cortex. Subsequent paralysis of skeletal muscles leads to variable degrees of motor impai ...

Loss of function is a standard approach to elucidate the function of a specific protein. Among these multiple strategies for silencing genes in living animals, genetic knockout in mice have been so far most frequently used. However, short hairpin RNAs (shRNAs) and microRNAs (miRNAs) deliver ...

Neural stem/precursor cells (NPCs) are defined according to their ability to self-renew and differentiate into all the major cellular types of the central nervous system (CNS). They can survive and proliferate in defined culture conditions as spherical aggregates (neurospheres) for a ...

Optogenetics is a technical methodology that allows direct light-based manipulation of genetically specified cells. Optogenetic methods have provided novel insights into the role of defined neuronal populations in brain function and animal behavior. An expanding palette of s ...

During the last decade, viral technologies have progressively been used in the studies of different aspects of brain functions. Relatively recently, viral techniques have been combined with optogenetics, which allows the study of modifications of the activity of different brain reg ...

Neurodegenerative diseases, such as Parkinson’s disease (PD), offer unique opportunities for the therapeutic application of viral vector gene transfer. In this chapter, we discuss the current state of viral vector preclinical development in nonhuman primate with special focus on v ...

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 ...

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 d ...

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 prot ...

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 d ...

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 favo ...

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 t ...

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 genera ...

Whole-mount in situ hybridization is the preferred method for detecting transcript distributions in whole embryos, tissues, and organs. We present here a sensitive fluorescent in situ hybridization method for colocalization analysis of different transcripts in whole embryon ...

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 devel ...