实验仪器使用技术

Transmission electron microscopy (TEM), a once popular research tool, is used less frequently now. However, the isolation of novel proteins in the past decade has led to renewed use of electron microscopy for the subcellular localization of these proteins. This chapter will describe the sta ...

The blood-brain barrier (BBB) is a unique dynamic regulatory interface situated at the border between the blood stream and the brain extracellular (or interstitial) fluid. As the “gatekeeper” to the brain, it determines the ability of drugs to gain entrance to brain extracellular fluid and re ...

The first report of blood-brain barrier (BBB) permeability by Paul Ehrlich (1) involved the use of the exogenous tracer Coerulean-S as described in Chapter 6. Over the years, tracers of different sizes were introduced to study the permeability properties of normal cerebral vessels in physio ...

Many techniques have been developed to study transport across the blood-brain barrier (BBB). One that offers particular advantage is the in situ brain perfusion technique. The primary objective of the in situ brain perfusion technique is to take over the circulation to the brain via direct inf ...

Efflux transport systems act as one of the detoxification systems at the blood-brain barrier (BBB) by eliminating xenobiotics and endogenous wastes from the brain into the blood (1–6). P-glycoprotein is a well-known example (1,2,5,6). It is a primary active transporter for hydrophobic neut ...

The blood-brain barrier (BBB) serves to regulate passage of solutes and water between circulating blood and brain extracellular fluid (1,2). It is composed of endothelial cells that line brain capillaries and exhibit selective transport properties. Cerebral endothelial cells are u ...

Cellular barriers in the central nervous system (CNS) present a formidable challenge in the delivery of drugs to the brain. These barriers include the blood-brain barrier (BBB) and the blood-cerebrospinal fluid barrier (BCSFB). Of these, given surface area and diffusional distance cons ...

The first successful isolation of cerebral microvessels from rat brain some 30 years ago opened the way to the development of in vitro models of the blood-brain barrier (BBB), which have since contributed considerably to the understanding of the physiology, pharmacology, and pathophysi ...

The ability to isolate and culture endothelial cells derived from brain microvessels has led to a considerable increase in our understanding of the biology of these cells over the last two decades. Most of these in vitro systems are derived from various animal sources and provide a valuable tool f ...

Primary vascular endothelial cell cultures provide powerful systems to investigate the molecular architecture and regulation of the blood-brain and blood-retinal barriers. Most investigators agree that in vitro models of endothelial cells alone do not completely recapitu ...

Several groups have reported the isolation and in vitro pharmacological characterization of brain microvessel endothelial cells (BMECs) of various origins: bovine, porcine, murine, or human (1–3). Brain capillaries are almost completely ensheathed by astrocyte processes, wh ...

The protein truncation test (PTT) (1–3), also known as the in vitro coupled transcription/ translation synthesis assay, was designed as a tool to detect mutations that lead to premature translational termination. It was originally developed to screen for mutations in the dystrophin (DMD) g ...

Prion disease represents a group of transmissible neurodegenerative disorders that include scrapie in sheep and goats, bovine spongiform encephalopathy (BSE), and Creutzfeldt-Jakob disease (CJD), Gerstmann-Str�ussler-Scheinker syndrome, fatal familial insomnia, a ...

The ability to generate antibodies that recognize a given protein relies on that protein’s “antigenicity”, i.e., its ability to appear foreign to the host immune system. Thus, proteins that are highly conserved among species are often poor antigens when used for immunization (1). This lack of imm ...

Tuberous sclerosis complex (TSC) is an autosomal dominant disorder characterized by the development of widespread hamartomas affecting most organs including the brain, heart, skin, kidney, lung, and eyes. Involvement of the brain is associated with some of the most severe clinical pro ...

Protein studies are of utmost importance for enhancing our understanding of genotype:phenotype correlations, as well as for diagnostic purposes. This is particularly true for the study of neuromuscular diseases, where defects in protein expression directly contribute to the eth ...

The use of formaldehyde as a reagent for fixing tissue was first identified over a century ago (1), and because of its efficacy in a broad range of situations, at varying concentrations and with virtually all tissues, it has remained one of the most widely used fixatives. Formaldehyde is a reactive elec ...

The application of in situ hybridization for the detection of neuropeptide mRNA has made a very significant contribution to the understanding of fundamental mechanisms of gene expression in neural tissues. First, as a powerful tool for analyzing the pattern of spatial and temporal expre ...

Ion-pair reversed-phase liquid chromatography (LC), as introduced by Johansson et al. in 1978 (1) is still the most widely used separation technique for analysis of the biogenic amines. It is usually coupled with electrochemical detection (ECD). The use of microbore LC was introduced by Scott ...

Microdialysis is a well-established sampling method for collecting neurotransmitters from the brains of freely moving rats (1; see also Chapter 19). To measure levels of amino acid transmitters in the microdialysates requires very sensitive methods of analysis owing to the subpicog ...