实验仪器使用技术

With the advent of modern chromatographic instrumentation and improved sequencing methods that allow for isolation and characterization of minute amounts of biological material, a host of biologically active peptides are now known. More recently, the dramatic development of ge ...

Recently there has been an explosion of information on peptides that may act as neurotransmitters or neuromodulator substances in the brain and periphery. Many investigators interested in the effects of neuropeptides have made use of radioligand binding techniques to identify pot ...

During the course of the last decade a considerable number of biologically active peptides has been identified in the nervous system; several of these recently discovered peptides fulfill the criteria for a neurotransmitter function (H�kfelt et al., 1980a); Snyder, 1980; Bloom, 1981; Krie ...

Numerous peptides have been localized with immunocytochemical techniques in neuronal cell bodies, axons, and nerve endings of the central nervous system (CNS). The widespread distribution of pepndes within the brain and spinal cord implies that these substances may function under p ...

The role of protein phosphorylation in regulating and integrating the activity of the nervous system has become widely appreciated during the last few decades. Both extracellular signals (e.g., neurotransmitters, hormones, growth factors, and electrical activity of the nerve cell ...

Nerve terminal depolarization leads to Ca2+ influx and the exocytotic release of neurotransmitters (Jessell and Kandel, 1993; Kelly, 1993). Elucidation of the regulatory influences on this primary function of the nerve terminal is of fundamental importance in understanding synap ...

Protein phosphorylation/dephosphorylation systems have evolved as a primary means through which cellular processes are regulated posttranslationally. Indeed, most transmembrane signals exert their biological effects, at least in part, by altering the balance of protein ...

Over the past four decades, studies directed toward the elucidation of mechanisms involved in the hormonal regulation of metabolism have burgeoned into the field we now know as cellular signal transduction. Both then and now, the role of protein phosphorylation has been central to these inv ...

This chapter describes the types of protein methylation that might be confronted in investigations of the nervous system, and summarizes technical approaches that have proven useful in elucidating the chemistries and identities of methylated proteins. Several distinct protein ...

Following translation, many proteins undergo further modifications that can dramatically affect both their physical properties and biological function (Wold and Moldave, 1984; Freedman and Hawkins, 1985; Harding and Crabbe, 1992). These posttranslational modifications a ...

Reversible modifications of cellular proteins often serve as key steps in signal transduction and effector pathways, and the investigation of these modifications has been a principal route of access to signaling mechanisms. Ten years ago, for example, tyrosine phosphorylation was an ...

Generally, genomic DNA is used either for the construction of genomic libraries or for Southern blot analysis. For several reasons, it is frequently preferable in the field of invertebrate neurobiology to screen, at least initially, genomic rather than complementary DNA (cDNA) librari ...

Neuronal voltage-activated Ca2+ channels have received consider- able attention from scientists because of the central role played by Ca2+ in the transduction of electrical activity to chemical signals. The importance of neuronal electrical activity resulting in a rise in intra- cel ...

Conventional agarose gel electrophoresis is a widely used technique for the analysis of many kinds of biological molecules, including fragments of DNA. It has one major limitation, namely its inability to resolve DNA fragments of greater than approx 30 kb in length. In human genetics, the enor ...

It is generally agreed that ion channels are operationally composed of two functional domains: The selectivity filter comprises the part of the protein that determines which ion may pass and which is retained; the gate determines under which conditions the selected ions may pass. Named acco ...

Muscle contraction is affected by the rapid release of Ca2+ ions through Ca2+- conducting channels localized in an intracellular mem- brane compartment, the sarcoplasmic reticulum (SR). The rabbit skel- etal muscle SR Ca2+ release channel has been identified as a high-affinity receptor for ...

Calcium channels are an essential part of the cellular signal trans duction system, since they produce changes in cytosolic calcium. Three types of voltage-dependent calcium channels (T, L, and N channels) have been identified by electrophysiological and pharmacological tech- niqu ...

Purification of receptor proteins has always been an important and challenging task in advancing the structural characterization and also in providing sequence data for the molecular cloning of the recep- tors. Some of the difficulties in the purification of receptors are:

A variety of biochemical and physiological studies in recent years have made it clear that the brain possesses specific receptors and puta- tive signal transduction pathways for insulin and the insulin-like growth factors (IGF-I and IGF-II). These observations have challenged tradi- t ...

This chapter is designed to serve as a guide for neurobiologists interested in studying neurotransmitter receptors and ion channels expressed in the oocytes of the African clawed frog Xenopls laevis. The emphasis is on routinely employed methods, based mostly on the experience gained in o ...