神经生物学

The α1B-adrenergic receptor (α1B-AR) is a G-protein-coupled receptor that plays a key role in the sympathetic regulation of a variety of physiological processes, such as cardiac and smooth muscle contractility, contraction of the spleen, liver glycogenolysis, melatonin secretion ...

Neurotransmitters act on cell-surface receptors to produce a wide range of effects on target cells (1). These effects include modulation of ion channels and changes in cellular electrical properties. Deciphering the intracellular molecular pathways by which receptor activation ...

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 ...

In order to isolate a single gene, phage or cosmid libraries can be screened by the conventional technique of hybridization as described by Sambrook et al. (1) using end-labeled oligonucleotide probes or gene-specific probes. The probes are labeled either by nick translation, end labeling, or ...

Many drugs and hormones interact with plasma membrane receptors to induce changes in the production of intracellular second messengers. The first such messenger to be identified was adenosine-3′,5′ cyclic monophosphate (cAMP), discovered by Sutherland and Rall in the late 1950s (1). Ch ...

It is believed that binding of agonist to a G-protein coupled receptor (GPCR) induces a set of structural changes in the tertiary structure of the receptor that can be recognized by the associated G-protein α-subunit. Many different methodological approaches have been applied over the years ...

The catecholamines epinephrine and norepinephrine produce a wide variety of physiological responses in target cells. Some of these are mediated by β-adrenergic receptors (β-ARs) through the stimulation of adenylyl cyclase and the production of cyclic AMP as first demonstrated by Su ...

Most membrane functions are mediated by enzymes and specific receptors that bind small molecules: substrates, inhibitors, modulators, hormones, and neurotransmitters. At the molecular level, polypeptide chains that contribute to the binding site of a given receptor can be identi ...

The power of polyacrylamide gel electrophoresis in resolving individual proteins in complex mixtures permitted the application of follow-up techniques to identify the separated components. A popular follow-up technique is immunoblotting (often referred to as Western blot ...

Adrenergic receptors mediate the central and peripheral actions of norepinephrine and epinephrine. Both of these catecholamine messengers play important roles in the regulation of diverse physiological systems and are widely distributed throughout the body. Agonists and an ...

Adrenergic and other G-protein-coupled receptors (GPCRs) are transmembrane molecules with complex secondary and tertiary organization intended to generate the binding pocket within the transmembrane loops (1). Although there are numerous differences in the primary amino a ...

The use of targeted gene disruptions, or knockouts, has become commonplace in many basic research laboratories. In its most common application, this technique enables the researcher to disrupt expression of a specific gene product selectively. This approach has been particularly us ...

Ablation of the mRNA of a targeted protein by the use of antisense DNA and RNA provides degrees of freedom not available in many other strategies to suppress or eliminate gene products (1–3). Numerous examples exist demonstrating the utility of the antisense DNA/RNA strategy for study of signali ...

There are several methods currently available for transfection of DNA into mammalian cells. These include transfection with calcium phosphate (1), diethylaminoethyl (DEAE)-dextran, cationic liposomes, nonliposomal lipid compounds, and electroporation (2). Each of these m ...

A large number of receptors for neurotransmitters and hormones have been identified by pharmacological studies and by molecular cloning. These receptors are divided into homologous families, which consist of a number of even more homologous subtypes. At least nine distinct adrener ...

G protein-coupled receptors (GPCRs) are among the most important targets for pharmacological compounds used in humans. During the last 10 years, more than 1000 genes coding for GPCRs, representing approx 0.1% of the human genome, have been identified, of which at least 200 have been demonstrat ...

As in many other fields of research in biology, the technology of expressing cloned genes in eukaryotic cells has become an increasingly important method for the study of adrenergic receptors (ARs). As developed in Chapter 14, substantial expression of adrenergic receptors can be achieved ...

Primer extension is often used to map the 5′ end of RNA (1). A single-stranded, end-labeled DNA primer is hybridized to RNA first. Using an RNA-dependent DNA polymerase (reverse transcriptase ) and nonradioactive deoxynucleotides, the primer is extended to yield cDNA. The cDNA is then analyzed on a s ...

The functionally diverse group of G-protein-coupled receptors (GPCRs) is a superfamily of membrane receptors. They include receptors for many different signaling molecules, such as peptide and nonpeptide hormones, neurotransmitters, chemokines, prostanoids, and protei ...

The quantification of adrenergic receptor (AR) mRNAs is an important tool in the study of the physiological and pathophysiological regulation of these receptors. Alterations of the levels of these mRNA represent one of the many mechanisms that regulate receptor signaling (1,2). Such al ...