实验仪器使用技术

The term “kindling” is a particularly apt metaphor when used to describe the phenomenon discussed in this chapter. In a manner similar to that in which small burning twigs ultimately produce a large fire, repeated subconvulsive stimulation of the brain by electrical current or pharmacolog ...

The purpose of this review is to present an overview of various experimental models of chemically induced seizures. It is not intended to be an exhaustive review of all chemical models, but rather a “how to” description of some of the more popular models that have yielded important data concerning me ...

In humans, deficiency of thiamine (vitamin B,) is associated with two well-defined neurological disorders: a mixed sensorymotor neuropathy and the Wernicke-Korsakoff syndrome. Disturbances of thiamine metabolism may also play a role in other human diseases, such as infantile suba ...

Adrenergic receptors (ARs) belong to the family of G-protein-coupled receptors and can be classified into three subfamilies, with each one having at least three distinct subtypes (1). Although all three subfamilies have similar affinity for the endogenous ligands norepinephrine and ...

α2-Adrenergic receptors (α2-ARs) are G-protein-coupled receptors that mediate the inhibitory effects of norepinephrine and epinephrine in the peripheral and central nervous system (CNS). In the CNS, these receptors have been implicated in the regulation of analgesia, memory, and c ...

AA is the precursor for prostaglandins, eicosanoids, and the endogenous cannabinoid ligands, such as anandamide (1). The major pathways that lead to the liberation of free AA from phospholipids are shown in Fig. 1. Phospholipase A2 (PLA2) provides a direct pathway for AA release and is the most comm ...

Ever since the initial discovery of the structurally homologous G-protein-coupled receptors, the search has been ongoing for identifying suitable expression systems. Such systems should be easy to use, inexpensive, and appropriate for mass production. In addition, these systems s ...

The redox modulatory sites of the NMDA receptor consist of critical cysteine residues, which when chemically reduced, increase the magnitude of NMDA-evoked responses. In contrast, after oxidation, NMDA-evoked responses are decreased in size. In recent years, as endogenous sources of ...

Phage-displayed random peptide libraries have been used to identify novel agonists and antagonists to a wide variety of receptors. Among these are a family of peptides that bind to the NMDA receptor subunit NR1 and non-competitively inhibit receptor channel activity (1). The peptide libr ...

There are two approaches to evaluation of the effects of drugs on steady-state ligand-gated receptor function: (1) concentration-response curves derived from whole-cell recordings and (2) analysis of single-channel recordings. Whole-cell currents are the average response of a la ...

Glutamate receptors are large integral membrane proteins that belong to the class of ligand-gated ion channels. They constitute a family of receptors that are activated by binding of the neurotransmitter l-glutamate. Functional receptors are formed by assembly of several subunits a ...

Neurotransmitter-gated channels, such as the NMDA receptor, are oligomeric membrane proteins assembled from multiple homologous subunits organized around a central ion-conducting pore (1,2). A major effort is directed toward understanding the structural features that det ...

A prominent current paradigm in the discovery of potential drugs directed toward therapeutic targets relies on the combination of brute force and specificity. The increasing number of entities in pharmaceutical company compound libraries, frequently surpassing 106, dictates ...

The NMDA subtype of excitatory glutamate receptor is a multisubunit, fast-acting, ligand-gated cation channel with a high permeability for Ca2+. NMDA receptors have been shown to be of importance in physiological processes, such as long-term potentiation, and also in pathophysiolog ...

Foreign gene transfer and expression in the cells are among the most important techniques for determining the characteristics and function of cloned genes. Glutamate receptor genes have been expressed by RNA injection of Xenopus oocytes or transfection of mammalian cells, such as HEK, C ...

Xenopus laevis oocytes have contributed greatly to the study of glutamate receptors. The isolation of the first cDNA clone for a glutamate receptor channel, GluR1, was achieved by utilizing an oocyte expression cloning system (1). In 1991–1992, three groups reported the isolation of cDNA cl ...

NMDA receptors are ligand-gated cation channels that are activated by glutamate and glycine. They are distributed throughout the mammalian CNS, and play an important role in normal development and plasticity of the CNS, and likely in such specialized functions as memory and learning. Pro ...

This chapter covers the major immunocytochemical methods used in our laboratory for NMDA receptor localization. There are three methods: pre-embedding immunoperoxidase for light microscopy (LM), the same for electron microscopy (EM), and postembedding immunogold (1–5). Gener ...

NMDA receptor is a ligand-gated ion channel receptor composed of multiple subunits encoded by at least five genes and their splice variants (1). Functional channels can be produced with only the NR1 subunit. These single-subunit channels, exhibit many of the key properties found in native cha ...

The quantification of drug interaction with NMDA receptors has been greatly facilitated by the use of ligand binding assays. The first assays for this receptor measured NMDA-sensitive glutamate binding (see 1 for review). However, glutamate has relatively low affinity for the receptor, ...