神经生物学

Excitatory and inhibitory neurotransmission mediated by glutamate and GABA, respectively, plays a major role in generation of seizures. So far, emphasis has been placed on the GABA system in attempts to develop antiepileptic drugs. Tiagabine, a selective inhibitor of GABA transporter 1 ...

Neuronal networks cultured on microelectrode array (MEA) neurochips provide a testing platform for neuroactive compounds such as neurotoxicants and neuropharmaceuticals. Electrical network activity is recorded, quantified, characterized and classified at the level of ...

Studies of ion channel pharmacology have witnessed considerable developments during the past half a century. Whereas voltage clamp techniques were applied to neuropharmacology for the studies of some chemicals in the late 1950s, it was not until 1960s that cellular neuropharmacolo ...

GABAA receptor (GABAAR) constitutes the main inhibitory receptor of the central nervous system. Due to the wide distribution and activity of its main neurotransmitter agonist, the γ-aminobutyric acid (GABA), its pharmacology has been thoroughly studied, given rise to the developme ...

This chapter describes the materials and methods necessary to generate human induced pluripotent stem cells (iPSCs) from primary human fibroblasts and direct their differentiation into neural progenitor cells. Application of such methods is an emerging model for the study of neuro ...

The relation of autolysis to calpain activation has a long and controversial history. Early on, it was realized that calcium-induced calpain activation is accompanied by autolytic truncation of the enzyme in vitro and that sensitization to calcium occurs at the same time (1). The issue of the me ...

The Ca2+ binding properties of calpain are of great interest, both biochemically in the wider context of EF-hand proteins, and physiologically, in the context of calpain regulation. There are two major parameters which one might wish to measure: the actual number of binding sites (n) and the bindi ...

Calpain activity can be assayed by a variety of methods which almost invariably rely on measuring the liberation of trichloroacetic acid (TCA)-soluble peptides from a protein substrate, usually casein. The soluble peptides are then measured either by their absorption at 280 nm, or by their r ...

Calpains play important roles in numerous physiological and pathological processes (1,2) by catalyzing the limited proteolysis of a wide variety of protein substrates. The activity of calpain toward a specific substrate is regulated not only by calcium, but also by numerous other facto ...

Many of the early studies on the localization of calpain described a cytosolic distribution. However, it is becoming clear that calpain can play an essential role in regulating signal transduction across members of the integrin family of adhesion receptors and that in order to do this it is recru ...

There is now a considerable literature suggesting that the calpain/calpastatin system may be involved in myogenesis (1–7). Primary myoblasts in culture at first proliferate, but then usually cease to divide, in response to complex signals. The cells then migrate, align, and fuse to form the mu ...

A major area of calpain research involves the development of techniques to assess calpain activation in cells. Knowing physiologic conditions that allow the various calpain isoforms in cells to become activated should provide valuable information about their roles in cell functio ...

Cell membrane fusion is a ubiquitous cellular process, mediating such phenomena as fertilization, muscle development, certain viral infections, and giant cell formation from macrophages. For membrane fusion to occur, the cell membranes must attach to each other, usually by means of in ...

Calpain is a calcium-activated neutral proteinase present in all mammalian tissues thus far studied (1,2). Ubiquitous calpain exists as μ-calpain and m-calpain isoforms which require μM and mM calcium levels for activation, respectively. In the central nervous system (CNS) μ-calpain is ...

Identification of the preferred brain substrates of calpain in vivo is not an easy task. The first difficulty is that, given enough time, almost any protein will be degraded when incubated in the presence of calpain and Ca+ in a test tube. A second major problem is the lack of completely specific calpain in ...

The calpains are found ubiquitously in mammalian cells and are activated following various central nervous system (CNS) insults, including ischemia (1), spinal cord injury (2) and traumatic brain injury (TBI) (3–5). Nonerythroid α-spectrin is a submembrane cytoskeletal protein and p ...

The preparation of freshly isolated rat renal proximal tubules in suspension has been utilized by our laboratory for more than 10 yr and reported in numerous publications (1–10). This method of tubule isolation by collagenase digestion and Percoll centrifugation has recently been used by ...

The general assay for proteolytic activity using casein as a substrate was described in 1947. A modification using azocasein has been described, in which digestion by calpain results in colored peptides soluble in trichloroacetic acid (TCA), so that the intensity of the color in the supernat ...

Although considerable information about calpain has been obtained from studies using isolated enzyme preparations, this does not permit the dynamic assessment of intracellular protease activity during changes in intracellular Ca2+ and pH. Intracellular calpain activity is ...

Ischemia-reperfusion (I/Rp) injury to the liver occurs after liver transplantation, shock or in surgical procedures in which vascular supply to the liver is temporarily abrogated. The extent of I/Rp-mediated injury is dependent on several factors including duration of ischemia, and in ...