神经生物学

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, ...

The term pyridoxine is generally used to refer to the group of naturally occurring pyridine derivatives represented by pyridoxine (pyridoxol), pyridoxal, and pyridoxamine, with similar physiological actions. They are referred to as vitamin B6 vitamers. Pyridoxine is used synonym ...

Hereditary hyperammonemias can be defined as a group of inborn errors in which the altered gene product would adversely effect the nitrogen metabolism of the affected individuals, causing an abnormal accumulation of ammonia in the blood. These altered gene products primarily include t ...

Quantification of radioligand binding by exposing labeled tissue sections to phosphor screens in cassettes is similar to autoradiography using radiation sensitive film (1) (see Chapters 5 and 7). The major advantage of phosphor screens over film is the greatly increased sensitivity ...

There are currently several screening methods for the detection of point mutations, such as single-stranded conformation polymorphism, heteroduplex analysis, denaturing gradient gel electrophoresis, and chemical cleavage. These are powerful tools for the identificati ...

Mutations in the parkin gene have been shown to be responsible for a substantial number of cases of autosomal recessive early onset parkinsonism (AR-JP, PARK2, OMIM 602544) worldwide (1–4). The gene on chromosome 6q25.2-27 consists of 12 coding exons with an open reading frame of 1395 bp. The gene is est ...

Huntington’s Disease (HD) is an autosomal dominant neurodegenerative disorder with an incidence of 1/10,000. The disease is characterized by involuntary choreic movements, psychiatric disorders, dementia, and death within 15-20 years. When the HD gene was cloned in 1993, it was discov ...

Fragile X syndrome, caused by the loss or diminution of the FMR1 (FRAXA - chromosomal locus Xq27.3) encoded protein, FMRP, results in mild to moderate mental retardation as its hallmark. Patients with the syndrome often vary dramatically in presentation with a range of intellectual and behavi ...

Ligand binding to receptors is a key step in the regulation of cellular function by neurotransmitters, hormones, and many drugs. Accordingly, this initial event in ligand action is important for understanding disease and designing new drugs. A large body of experimental data describing r ...

G protein-coupled receptors (GPCRs) make up the largest and most diverse family of transmembrane proteins and respond to a wide variety of stimuli including biogenic amines, peptides, bioactive lipids, hormones, and light (1,2). Agonist binding to these receptors activates intracel ...

Historically, the G protein-coupled receptor (GPCR) protein family has proven to be an extremely tractable target class (1). It is estimated that approximately one-half of all drugs currently marketed exert their actions, either directly or indirectly, via GPCRs (2). Given the potential c ...

Scintillation proximity assay (SPA) is a homogeneous assay technology (1) which is bead-based and removes the need for a filtration step to separate bound from free ligand in a receptor-binding assay. The principle of the technology is shown in Fig. 1 .