细胞功能测定

This chapter describes in vitro methods for studying the Blood-Brain barrier. These methods include a cell line and isolated brain microvessels. The rat brain endothelial cell line 4 (RBE4) express many properties that are expressed by brain endothelial cells in vivo. Tissue culture meth ...

Lipid rafts are dynamic structures made up of proteins and lipids that float freely within the liquid-disordered bilayer of cellular membranes and have the ability to cluster to form larger, more-ordered platforms. These clustered structures have been identified in all cell types and have ...

Epithelial cells at all mucosal surfaces are potentially apposed to bacteria, particularly in the intestine. It is established that intestinal epithelial cells (IECs) represent an important barrier between lamina propria cells and the potentially harmful lumenal contents. In ad ...

Adhesive interactions between cells regulate tissue integrity as well as the process of inflammatory cell recruitment. Such intercellular interactions are regulated by adhesion receptors and can be homotypic, that is, between cells of the same type, for example, between adjacent en ...

Disrupted epithelial cell junctions are a hallmark of numerous disease processes. The signaling mechanisms regulating barrier function and re-establishment of intact junctions after injury and during development are complex and tightly regulated. We have shown that heterot ...

Models to study molecular, biochemical, and functional responses in vitro generally incorporate an individual cell type or group of cells organized in a random fashion. Normal physiological responses in vivo require that individual cell types be oriented in an organized fashion with t ...

Interactions between the vascular endothelium and polymorphonuclear leukocytes (PMNs) are central to PMN emigration into inflamed tissues, and to neutrophil–endothelial crosstalk pathways that modulate inflammatory responses and vascular barrier function. For exam ...

The interactions between egg and sperm are among the most fascinating in cell biology. These interactions include Cell-Cell adhesion and then membrane fusion between the two gametes. This chapter details the experimental methods used to for gamete culture and in vitro fertilization us ...

Mucosal tissues undergo contraction and relaxation on a continuous basis. In its normal state, the pliable intestinal tract is characterized by a rhythmic pattern of contractions controlled by its intrinsic neuronal innvervation. In chronic inflammatory diseases such as Crohn’s d ...

An essential requirement for adequate organ performance is the formation of permeability barriers that separate and maintain compartments of distinctive structure. The endothelial cell lining of the vasculature defines a semipermeable barrier between the blood and the inter ...

The preparation of mammalian mitochondrial ribosomes in sufficient quantities for biochemical studies is best done beginning with whole tissue rather than from cells in culture. This issue arises because of the low abundance of these ribosomes in cells, making their isolation a challe ...

Blue native gel electrophoresis (BNGE) is a powerful tool for analyzing native protein complexes from biological membranes as well as water-soluble proteins. It can be used for determining relative molecular masses of protein complexes and their subunit composition and for the detec ...

Import of proteins into mitochondria is a pivotal process in the biogenesis of mitochondria. Only about 1% of the 1000–2000 different proteins constituting the mitochondrion are encoded in the mitochondrial deoxyribonucleic acid (DNA). All others are specified by nuclear genes. They a ...

In plants, the majority of mitochondrial and chloroplast proteins are nuclear encoded, synthesized on cytosolic polyribosomes, and then imported into the organelle. Most of the nuclear encoded precursor proteins contain an N-terminal extension called signal or targeting pepti ...

Blue native polyacrylamide gel electrophoresis (BN-PAGE) employs the dye Coomassie for the labeling of proteins and protein complexes under native conditions. Electrophoresis under native conditions subsequently allows resolution of proteins and protein complexes acc ...

Iron-sulfur (Fe-S) clusters are cofactors of many proteins that are involved in central biochemical pathways, such as oxidative phosphorylation, photosynthesis, and amino acid biosynthesis. The assembly of these cofactors and the maturation of Fe-S proteins require complex cell ...

Mitochondria are dynamic organelles with activities that adjust to altering physiological conditions and variable metabolic demands. A conserved proteolytic system present within the organelle exerts essential functions during the biogenesis of mitochondria and ensu ...

This chapter describes a luciferase-based protocol to measure adenosine triphosphate (ATP) production in isolated mitochondria of Trypanosoma brucei. The assay represents an excellent method to characterize the functionality of isolated mitochondria. Comparing the ATP p ...

The adenosine triphosphate (ATP) synthase (F1-F0 complex) of the mitochondrial inner membrane is responsible for making nearly all of the ATP utilized by eukaryotic organisms. The enzyme is an oligomer of more than 20 different subunits, 14 of which are essential for its catalytic activity. T ...

Mitochondria not only are a source of reactive oxygen species (ROS) but also are sites of oxidative damage. In plants, mitochondria must normally operate when there are high levels of ROS produced during photosynthesis and photorespiration. These levels are further enhanced during biot ...