细胞功能测定

Virtually all eukaryotic cells can grow in a polarized fashion in response to external signals. Cells can respond to gradients of chemoattractants or chemorepellents by directional growth, a process referred to as chemotropism. The budding yeast Saccharomyces cerevisiae underg ...

Electric fields were measured at human skin wounds over one and half centuries ago. Modern techniques have verified and greatly extended our understanding of the existence of endogenous wound electric fields. In virtually all wounds studied, disruption of an epithelial layer instant ...

Dictyostelium slugs are able to respond to environmental stimuli in an extremely sensitive and efficient way. This enables a slug to migrate to more favourable locations for formation of fruiting bodies and dispersal of spores. Phototaxis is a readily assayed phenotype and reflects the in ...

Motile microorganisms react to a host of external stimuli, including light, gravity, the magnetic field of the Earth as well as thermal and chemical gradients, in their habitat in order to select a niche suitable for survival and reproduction. Several forms of light-induced behavior have been d ...

The task of developing and simulating computational models of signaling networks for eukaryotic chemosensing confronts the modeler with several challenges: (1) The stimuli that initiate the cellular responses one wishes to study are provided by extracellular concentration gr ...

When stimulated by chemoattractants, eukaryotic cells respond through a combination of temporal and spatial dynamics. These responses come about because of the interaction of a large number of signaling components. The complexity of these systems makes it hard to understand without a ...

Many eukaryotic cells move in the direction of a chemical gradient. Several assays have been developed to measure this chemotactic response, but no complete mathematical models of the spatial and temporal gradients are available to describe the fundamental principles of chemotaxis. ...

For decades, Dictyostelium discoideum has been an efficacious and attractive model system for the study of cell motility, primarily because cells become highly motile during the transition from growth phase to aggregation competence and because the haploid genome is readily amenab ...

Microorganisms have various mechanisms at their disposal to react to (changes in) their ambient light climate (i.e., intensity, color, direction, and degree of polarization). Of these, one of the best studied mechanisms is the process of phototaxis. This process can be described as a behavior ...

B-cell activation initiates antibody responses against pathogens. Recent imaging of B cells in vivo shows that B cells move rapidly through lymphoid tissues to search for antigens captured on the surfaces of antigen-presenting cells. Recognition of antigens by the B-cell antigen rece ...

In this chapter, we describe methods to monitor signaling events at the single-molecule level on the membrane of living cells by using total internal reflection fluorescence microscopy (TIRFM). The techniques provide a powerful tool for elucidating the stochastic properties of sign ...

Since directed movement toward an extracellular chemoattractant requires rapid and continuous reorgani�zation of the actin cytoskeleton to form complex structures such as a protruding lamellipodium, it is of great interest to analyze and understand the individual contribut ...

This chapter will focus on responses that the chemoattractant cyclic AMP elicits in the motility system of Dictyostelium. These cells can be permanently transfected to express cytoskeleton-associated proteins tagged with fluorescent proteins. Multiple proteins that are dis ...

How a eukaryotic cell translates a small concentration difference of a chemoattractant across the length of its surface into highly polarized intracellular responses is a fundamental question in chemotaxis. Chemoattractants are detected by G-protein-coupled receptors (GP ...

Dictyostelium discoideum is a useful cell model for studying protein–protein interactions and deciphering complex signaling pathways similar to those found in mammalian systems. Many of these interactions were analyzed using classical in vitro biochemical techniques. How ...

Platelet microparticles are submicron vesicles that can support thrombin generation on externalized negatively charged phospholipids. Increased numbers of circulating platelet microparticles have been investigated as the basis of hypercoagulability in a variety of p ...

Isolation of pure platelet samples from whole blood is crucial for the study of platelet gene expression. The main obstacles to overcome in order to successfully isolate platelets from whole blood include (1) platelet activation; (2) leukocyte and red blood cell contamination, and (3) time- ...

Platelets are anucleate cytoplasts that circulate in the bloodstream for approximately 9–11 days. Because they lack nuclei, platelets were considered incapable of protein synthesis. However, studies over the last decade have revealed that platelets use a variety of translational ...

Intravital fluorescence microscopy is increasingly used to measure experimental arterial thrombosis in large and small arteries of mice in vivo. This chapter describes protocols for applying this technology to detect and measure thrombi formed by: (1) ultrasound-induced ruptu ...

Platelet aggregation responses are conventionally assessed in cuvette-based systems using either isolated platelets or whole blood. Unfortunately, in vitro aggregometry poorly predicts in vivo functionality, since mediators derived from the vascular endothelium are ma ...