细胞技术

Growth factor receptors, such as the epidermal growth factor receptor (EGFR), stimulate a variety of signal transduction pathways upon binding a ligand molecule at the cell surface. Desensitization of signaling initiates when active receptors are recruited to clathrin-coated re ...

Recently the life cycles of receptor tyrosine kinases (RTKs) have become a focus of signal transduction research. Ligand-induced ubiquitination of RTKs followed by their internalization and degradation has, in particular, been extensively studied. This chapter describes the ba ...

All ligands of the epidermal growth factor receptor (EGFR) are made as membrane anchored precursors that can be proteolytically processed and released from the plasma membrane. This process, which is referred to as protein ectodomain shedding, is emerging as a key regulator of the function of ...

Oxidative stress (OS) is linked to the development of human diseases. Early identification of OS-associated diseases is essential in the control of their progression and treatment. Efforts have been undertaken to identify reliable endogenous markers, which correlate with the prog ...

The nitration of protein tyrosine residues represents an important posttranslational modification during development, oxidative stress, and biological aging. The major challenge in the proteomic analysis of nitroproteins is the need to discriminate modified proteins, us ...

During the last few years, reliable and simple tests have been proposed to estimate oxidative stress in vivo. Many of them can be easily adapted to automated analyzers, permitting the simultaneous processing of a large number of samples in a greatly reduced time, avoiding manual sample and reage ...

Microscopy studies of oxidative stress have often consisted of only immunocytochemical localization or only cytochemical localization studies. However, correlative studies on the same sections provide more useful data in interpreting oxidative and nitrosative stress. C ...

Atherosclerosis is associated with dysfunctional HDL, and oxidation of HDL is thought to give rise to HDL becoming dysfunctional. Lipoprotein oxidation represents a complex series of processes that can be assessed by various methods. In general, oxidation mediated by 1-electron or ra ...

High performance liquid chromatography/electron spin resonance/mass spectrometry (HPLC/ EPR/MS) analyses of radicals is performed for the reaction mixture of 13-hydroperoxy-(9Z,11E)-octadeca-9,11-dienoic acid (13ZE-OOH) with ferrous ions under an aerobic condition, ...

Mitochondrial superoxide (O2 •–) production is an important mediator of oxidative cellular injury and pathogenesis of many diseases such as myocardial ischemia/reperfusion. The O2 •– generated in mitochondria acts as a redox signal triggering cellular events including apoptos ...

Medicinal drugs do not always have clearly understood mechanisms of action, especially as regards psychiatric treatment. Identification of genes involved in drug resistance is an important step toward elucidating the genetic basis of disease and the molecular mechanism of drug act ...

Dictyostelium discoideum is a molecularly amenable host model system for several human pathogens, including Legionella pneumophila, Mycobacterium avium, Mycobacterium marinum, Pseudomonas aeruginosa, and Cryptococcus neoformans. Dictyostelium wild-type cells ...

Intercellular signaling mediated by cAMP plays a pivotal role in coordinating cell movement into aggregates at the early stage of Dictyostelium development when the extracellular level of cAMP periodically changes at 6- to 7-min intervals. We have shown that MAP kinase ERK2 is activated v ...

This chapter describes biochemical assays with which to analyze signal transduction from surface cAMP receptors via G proteins to the effector enzymes adenylyl cyclase, guanylyl cyclase, and phospholipase C. The cAMP-mediated formation of the second messengers cAMP, cGMP, and Ins(1 ...

Research on Dictyostelium has a long history, the trend of which has been to add cell biology, natural products, evolutionary biology research to the traditional studies of development. The methods presented here and the recent publication of the genome present vast new opportunities.

The chemotactic signal in Dictyostelium is a cAMP wave that is relayed over relatively large distances through a cell population during aggregation. Cells exhibit unique behaviors in response to the different spatial, temporal, and concentration components of the cAMP wave, suggest ...

A method for the integration of linear DNA into the Dictyostelium genome is described. Restriction enzyme-mediated integration, or REMI, involves the transformation of cells with a mixture of plasmid DNA, linearized with a restriction enzyme, along with a restriction enzyme that is capa ...

Dictyostelium slug phototaxis and thermotaxis are readily assayed phenotypes that reflect with great sensitivity and specificity the interactions of environmental stimuli with morphogenetic signaling systems controlling the collective movement of slug cells. Metho ...

Normally, vegetative Dictyostelium grow as haploid cells. Occasionally, two haploid cells fuse together during normal growth, forming a diploid cell containing both parental sets of chromosomes within a single nucleus. The diploid state is reasonably stable, and the growth, develo ...

While spontaneous differentiation is an undesired feature of expanding populations of embryonic stem cells, a variety of methods have been described for their intended differentiation into specialized cell types, such as the osteoblast or chondrocyte. Most commonly, different ...