细胞功能测定

In this chapter we review protocols for transient transfection of primary keratinocytes. The ability to transfect primary epidermal cells regardless of their differentiation status allows the biochemical and molecular characterization of multiple proteins. We review meth ...

Compared to primary keratinocytes, HaCaT cells are easier to transfect and yet still maintain at least some features of normal epidermal proliferation and differentiation. This chapter describes methods used in our laboratory to maintain HaCaT cells in an undifferentiated state and ...

Many eukaryotic cells can elicit intracellular signaling relays to produce pseudopodia and move up to the chemoattractant gradient (chemotaxis) or move randomly in the absence of extracellular stimuli and nutrients (random movement). A precise spatiotemporal regulation of Ras ...

Invadopodia are hair-like membrane protrusions projecting from the ventral side of the plasma membrane of tumor cells invading into an extracellular matrix (ECM). Formation of invadopodia and phagocytosis of partially degraded ECM is correlated with invasiveness of cancer cells. ...

Intravital two-photon imaging allows the observation of immune cells in intact organs of live animals in real time. Recently, several studies using two-photon microscopy have detailed the motility of mouse B and T lymphocyte within lymph nodes and have shown a dependence upon chemokine rec ...

A broad array of biological responses ranging from cell polarization, movement, immune and inflammatory responses, as well as prevention of HIV-1 infection, are triggered by the chemokines, a family of structurally related chemoattractant proteins that bind to specific seven-tra ...

Asymmetric localization of intracellular proteins and signals directs movement during axon guidance, endothelial cell invasion, and immune cell migration. In these processes, cell movement is guided by external chemical cues in a process known as chemotaxis. In particular, leuko ...

Zebrafish have emerged as a powerful model organism to study neutrophil chemotaxis and inflammation in vivo. Studies of neutrophil chemotaxis in animal models have previously been hampered both by the limited number of specimens available for analysis and by the need for invasive proce ...

Quantitative studies of chemotactic signaling require experimental techniques that can expose single cells to chemical stimuli with high resolution in both space and time. Recently, we have introduced the method of flow photolysis (Anal. Chem. 79:3940–3944, 2007), which combines mi ...

The primary function of HIV-1 binding to its chemokine coreceptors is to mediate fusion and viral entry. However, it has been known that this interaction also triggers a variety of signaling cascades. It is likely that the virus-mediated signaling events may facilitate viral infection in var ...

The involvement of calcium signalling during chemotaxis in Dictyostelium discoideum is well documented. Spatiotemporal increases of intracellular calcium (i) have been observed within seconds of stimulation with the chemoattractants folic acid and cAMP. This rise in i locali ...

Anton van Leeuwenhoek first observed bacterial motility in the seventeenth century, and Wilhelm Pfeffer described bacterial chemotaxis in the late nineteenth century. A number of methods, briefly summarized here, have been developed over the years to quantify the motility and chemo ...

Phosphoinositide (PI) signalling is one of multiple signalling cascades involved in chemotaxis in Dictyostelium discoideum. PI signalling comprises a complex interaction of multiple enzymes, each with multiple phospholipid substrates and thus products, often relying upon ...

Dictyostelium discoideum is one of the most facile eukaryotic systems for the study of chemotactic response to secreted chemical ligands. Dictyostelium grow as individual cells, using bacteria and fungi as primary nutrient sources; during growth, Dictyostelium moves direction ...

Chemotaxis is a highly coordinated biological system where chemoattractants trigger multiple signal transduction pathways which act in concert to bring about directed migration. A signaling pathway acting through PIP3, which accumulates at the leading edge of the cell, has been ex ...

During development a variety of cell types are guided by molecular concentration gradients to form tissues and organ systems. In the nervous system, the migration and neuronal pathfinding that occurs during development is organized and driven by “guidance cues.” Some of these cues are subs ...

We describe an in vivo invasion assay that enables the collection of invasive cells from the primary tumor. In addition to determination of the endogenous, unstimulated invasive properties of cells in vivo, the assay can take advantage of the chemotactic properties of cancer cells. Microne ...

Cell motility is a widely researched and clinically relevant process that has primarily been investigated using cell culture models. While these in vitro assays are useful in allowing for high-resolution analysis of cell movement, there will always be questions surrounding the physi ...

Correct distal tip cell (DTC) migration in the nematode C. elegans requires sensing soluble and matrix cues, remodeling extracellular matrix, and signaling through conserved integrin and netrin pathways. The DTC executes a complex path and coordinates its migration with the develop ...

The ability of cells to migrate directionally in gradients of chemoattractant is a fundamental biological response that is essential for the survival of the social amoebae Dictyostelium discoideum. In Dictyostelium, cAMP is the most potent chemoattractant and the detection, synt ...