实验仪器使用技术

Neurotoxicological testing is mainly based on experimental animal models, but several cell and tissue culture models have been developed to study the mechanisms of neurotoxicity. In general, cells of human origin are attractive alternatives to the animal models for extrapolation of ...

A major goal of our cellular toxicology research has been the identification of novel targets of organophosphorous compounds (OPs), to which end we have studied the effects of sub-lethal concentrations of OPs on morphological and molecular end points in differentiating mammalian cell ...

Cataract formation during preclinical drug safety assessment studies can be a devastating safety finding during drug development based on the stage at which these findings usually occur. The lens explant culture models offer an extremely versatile and simple in vitro model to screen co ...

Most neurodegenerative diseases culminate in cell death, although it is not uncommon for signs of dysfunction to precede cell death in humans and animal models. There is considerable evidence that neuronal and glial cell death during development occurs through apoptosis but whether ap ...

Inflammation contributes to a wide variety of brain pathologies. In this chapter methods are described for using microglia and astrocytes in culture to investigate inflammatory processes and inflammatory neurodegeneration, including the use of neuronal/glial co-cultures a ...

Oxidative stress is implicated as one of the major underlying mechanisms in a variety of human diseases. Reactive radicals, derived primarily from molecular oxygen, readily attack a variety of critical biological molecules, including DNA, cellular proteins, and lipids. Since lipid p ...

Carbonyl group formation on protein side chains is a common biochemical marker of oxidative stress and is frequently observed in a variety of acute and chronic neurological diseases including stroke, Alzheimer’s disease, and Parkinson’s disease. Given that proteins are often the imme ...

The selective loss of melanin-containing dopaminergic neurons in the substantia nigra pars compacta and locus coeruleus has motivated a large number of preclinical studies aimed at understanding the molecular mechanisms involved in the neurodegeneration of dopaminergic ne ...

The use of various in vitro systems is expanding dramatically not only in basic research, but also to meet regulatory requirements for chemicals and products of various kinds. Further significant developments are certain to result from the use of in vitro systems for high-throughput scree ...

The use of 13C and 15N labeled precursors in combination with adequate analytical tools makes it possible to study metabolic pathways in cultured neural cells. The most commonly used precursors are 13C labeled glucose, lactate, glutamate and acetate. For a dynamic evaluation of intermedi ...

Glutamine (Gln) plays an important role in satisfying brain metabolic demands and as a precursor for the synthesis of glutamate and γ-aminobutyric acid (GABA). In vitro cultured cell studies have shown that carrier-mediated Gln transport between astrocytes and neurons represents a key ...

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 ...