神经生物学

The zebrafish (Danio rerio) has been at the forefront of neurobiological research and is steadily gaining favor as a model organism for behavioral applications. The ease of handling, high yield of progeny, and efficient mode of drug delivery make this species a particularly useful model for be ...

Information is transmitted throughout the nervous system by chemical and electrical signals. The cacophony of neuronal signals does not end at the cell surface, but continues deep into cells via diffusible intracellular messenger molecules. The list of molecules that serve as intrac ...

The patch-clamp method was introduced in 1976 (Hamill et al., 1981; Neher and Sakmann, 1976). It has been widely used to study whole-cell and single-channel currents. In many cases, the experimenter needs to test how changing the pipet solution composition can affect membrane-channel function ...

“The determination of the three-dimensional structures of protein molecules showed for the first time in detail the construction of the molecular ’machines’ of the life cycle. If we want to learn how these ’machines’ work it is not sufficient only to know their construction. We actually have to see ...

The great advantage of patch clamp in cellular physiology is that it allows sensitive and reliable analysis of the electrical activity of cell membranes at the molecular level. Whole cell is the most popular variant of the patch-clamp technique. It is easy to obtain and it allows the use of intra- and ext ...

Brain slices have become an integral part of synaptic and cellular physiology since the pioneering studies by Henry McIlwain (Li and McIlwain, 1957; Yamamoto and McIlwain, 1966). The hippocampal slice preparation was first brought to the United States from Per Anderson’s lab-initially by ...

One of the main challenges for a better understanding of signaling mechanisms in the normal and diseased central nervous system (CNS) is to unravel the molecular basics of function on the cellular and systemic levels. Several important cellular processes, such as proliferation, differe ...

The purpose of this chapter is to describe the procedures used to record macroscopic currents from patches of biological membranes. Several variations of the classic patch-clamp recording procedure are available and described in other chapters of this book, however the procedure rep ...

As mentioned by Cole (1968) the connections between life and electricity have long held the attention of investigators, with observations dating from ancient examinations of the electric catfish Malapterurus in 2600 B.C to modern examinations of the structure/function relations ...

ADP-ribosylation of proteins was originally discovered in 1966 by Chambon et al. (1966), who detected polymers of ADP-ribose attached to protein substrates. Since that time, protein ADP-ribosylation has been identified in a diverse range of species, from bacteria to human, as well as within v ...

Phosphorylated tyrosyl residues in proteins are quantitatively much less abundant in normal cells than phosphoserines or phosphothreonines, yet tyrosine phosphorylation plays a crucial role in cellular regulation, including cell transformation by viral oncogenes and the ...

A relatively new proteomic technology platform known as stable isotope labeling of amino acids in cell culture (SILAC) has been developed to record quantitative changes in in vitro experimental systems. This method allows for monitoring of changes due to various experimental conditi ...

Recent evolution in proteomics approaches from two-dimensional gel electrophoresis to peptide-based “shotgun proteomics” methods has greatly enhanced the abilities of scientists to uncover expression changes among “low abundant” proteins. Shotgun proteomics methods ...

The current protocol describes in the first part the isolation and culture of neural stem and progenitor cells. In the second part, the two-dimensional gel electrophoresis of neural stem and progenitor cells is explained, consisting of sample preparation, first-dimensional isoelec ...

We describe a method for analyzing and sharing gene expression data using a Web-based system, “Gemma.” Gemma is designed to support meta-analysis in a collaborative framework. Gemma allows users to log on, upload their data, annotate and analyze it, and share the data with others. Users can then comp ...

Massive parallel sequencing will become the method of choice for transcriptome profiling. Two protocols have been developed to quantify level of expressions: full-length RNA sequencing (RNA-SEQ) and 3′ tag digital gene expression (DGE). We have studied the performance of 3′ tag DGE profi ...

The brain vasculature, and more specifically the microvasculature, represents a center of interest for the investigation of the brain. Indeed, this tissue at the interface between the circulatory system and the central nervous system (CNS) actively participates in the brain function ...

The development of a variety of high-throughput (HT) technologies in molecular biology has markedly extended its analytical power and has given rise to an exponential jump of accumulation of data. The technologies can be roughly sorted into the ones used to analyze the transcriptome and the p ...

The process of RNA amplification is a stepwise series of molecular manipulations intended to amplify transcriptomic signals from small quantities of starting materials, including single cells and homogeneous populations of individual cell types for microarray analysis and o ...

Microarray expression profiling of the nervous system provides a powerful approach to ascribe activities to genes involved in distinct phases of neural development and function. Expression profiling of neural tissues and cell lines requires isolation of high-quality RNA, ampli ...