神经生物学

The methods described in this chapter are designed for rapid and convenient separation and quantitation of calpain isozymes in cultured osteoblastic cells by modern semiautomated strong-anion exchange FPLC. The isozymes of the calpain-calpastatin system are abundant in osteo ...

There is no single affinity chromatography method that is universally effective for the purification of calpains. Dye affinity chromatography and calcium-dependent binding to immobilized substrates or inhibitors are the most commonly attempted affinity methods. Our labor ...

The purification of μ-and m-calpain on a medium to large scale from animal tissues such as bovine heart or skeletal muscle, and on a microscale from less than 0.5 g of tissue, is described in Chapters 1 and 2. These preparations are excellent for many purposes, but for some other purposes, for example for cryst ...

Calcium-dependent cysteine proteinases (CDPs or calpain-like proteinases) constitute a large family of related proteins in the tissues of invertebrate species. They vary in native mass from 59 kDa for lobster muscle CDP III to 520 kDa for octopus muscle CDP, and vary also in subunit composit ...

p94 (also called calpain 3, nCL-1, or CAPN3) is a calpain large subunit homologue, which is predominantly expressed in skeletal muscle (1–4). The mRNA level of p94 in skeletal muscle is at least 10 times higher than that for the conventional calpain subunits (1). However, the p94 protein is not easily dete ...

Methods for analyzing enzyme activity after electrophoresis in acrylamide gels have been described for a variety of proteins, including several proteases and kinases. The application of this approach to detect calpain activity by casein zymography was first described by Raser et al. in ...

A number of brain insults including traumatic brain injury (TBI) can result in excitotoxic consequences largely attributable to pathological increases in intracellular calcium (1,2). Loss of calcium homeostasis can result in activation of the calcium-dependent proteases, or ca ...

The use of human postmortem brain tissue in neurochemical and neuropharmacological research has received increasing attention over recent years. In fact, there is one work that, more than any other, can be identified as being responsible for the interest in this approach. It was Birkmayer and ...

Magnetic resonance spectroscopy (MRS) provides information that is rarely obtainable by other noninvasive means, or even by invasive methods using radioactive labels. For example, it provides the means to monitor in time and in space changes in various metabolic pools and allows one to thi ...

Cell culture has proven to be a very powerful approach in addressing neurobiological questions. The reasons for this include the ability to isolate the effects of specific variables on cells and, more importantly, to ask questions of a specific cell type in isolation of other cells. The power of cell ...

In the last twenty years, the availabihty of radioactively labeled compounds that bind with high affinity and selectivity to pharmacological receptors has enabled the direct measurement of such binding sites. After initial studies, carried out with tissue homogenates, membrane pr ...

The aim of the present review is to summarize the methods used to estimate the turnover of various neurotransmitters of low molecular weight in the intact animal brain First, some introductory remarks and definitions are given. The following description of the terms used is derived mainly from ...

The term fluorescence commonly refers to the phenomenon in which light from the ultraviolet (UV) spectrum is absorbed by a substance and emitted in the visible spectrum. Certain organic molecules absorb light energy, resulting in various interatomic bonds being raised to higher energy l ...

Investigations in vitro on the transport of biogenic amines have done much to expand our knowledge of neurotransmission in general, of the role of amines in nervous function, and of the actions of a wide variety of psychotropic drugs In this review, we will be describing techniques that are employed ...

Gas chromatography (GC) is a technique used to separate and quantify the components of a mixture of volatile compounds by partitioning them as vapors between two phases—a stationary and a mobile phase. The stationary phase is contained in a narrow tube (the column) through which the mixture of com ...

It is evident that an explosive growth has occurred over the past ten years in the high performance liquid chromatographic (HPLC) analysis of catecholamines, indoleamines, and the trace amines In part, this expansion can be attributed to a growing interest in the monoamines; however, the incr ...

The biogenic amines 5-hydroxytryptamme (5-HT; serotonin), dopamine (DA), noradrenaline (NA), adrenaline (A), and histamine (HA) are present in mammalian brain, in which they play fundamental roles as neurotransmitters. Concentrated in a few brain nuclei and tracts (Dahlstrom and Fux ...

One of the fundamental goals of neuroscience is an understanding of the relationship between neurotransmission and behavior. Although considerable information has come from methods such as push-pull perfusion, obtaining data about neurotransmitter release in behaving ani ...

Mass spectrometry (MS) is now widely used in the neurosciences to identify and quantify a variety of biogenic compounds. When used in combination with chromatography, i.e., packed column or high resolution capillary column gas chromatography (CC or HRGC) or thin layer chromatography (TLC ...

The so-called trace amines or noncatecholic biogenic amines have recently attracted a considerable amount of attention among neuropharmacologists and psychiatrists. Some of these amines, e.g., phenylethylamine (PEA) and tyramme, are suspected to play some yet undetermined role ...