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A variety of bioartificial liver support systems were developed to replace some of the liver’s function in case of liver failure. Those systems, in contrast to purely artificial systems, incorporate metabolically active cells to contribute synthetic and regulatory functions as well as ...

The use of cryopreserved hepatocytes has increased in the last decade due to the improvement of the freezing and thawing methods and has even achieved acceptance by the U.S. Food and Drug Administration for use in drug-metabolising enzyme induction studies. This chapter provides an overview ...

The accumulation of insoluble Aβ peptide aggregates in the brain is the diagnostic feature of Alzheimer’s disease. Identical deposits are seen in the elderly who are at risk for this disease. The formation of the approx 4 kDa Aβ peptide is implicated as a key component in the development of Alzheimer’s ...

This chapter presents fluorescence microscope assays that can be used to study microtubule (MT)-based movement and receptor-ligand sorting in vitro. The strategy is to isolate endosomes in a concentrated active form and store them in frozen aliquots for single use. Glass microchambers ...

Ectoplasmic specializations are testis specifi c intercellular adhesion junctions found in Sertoli cells. They are tripartite structures consisting of the plasma membrane of the Sertoli cell, a submembrane layer of actin fi laments and an attached cistern of endoplasmic reticulu ...

F0F1-ATP synthase catalyzes the synthesis of ATP using proton-motive force across a membrane. When isolated, the F1 sector, composed of five polypeptide chains with a stoichiometry of α3β3γδε, solely hydrolyzes ATP into ADP and phosphate, and is thus called F1-ATPase. Rotation of a shaft dom ...

Fluorescence resonance energy transfer (FRET) is a spectroscopic phenomenon that consists of long-range dipole-dipole interaction between two chromophores. This method can be employed to gain quantitative distance information on macromolecules. FRET is particularly us ...

To understand the interaction of kinesin and microtubules, it is necessary to study the three-dimensional (3D) structures of the kinesin-microtubule complex at a high enough resolution to identify structural components such as α-helices and β-sheets. Electron cryo-microscopy co ...

Kinesins are molecular motors that share a common structural core with myosins and G proteins and play diverse roles in organelle transport and cell division and movement. Kinesin motors use the chemical energy derived from ATP hydrolysis to generate force for moving on the microtubule tra ...

The construction of a sensitized mutant unconventional myosin is an excellent method for determining the function of the individual myosin against a background of related myosins with partially overlapping functions. In this chapter, we outline the steps involved in sensitizing my ...

Kinesins, a superfamily of microtubule motor proteins, are implicated in regulating a number of fundamental cellular and developmental processes including intracellular transport of vesicles and organelles, mitotic and meiotic spindle formation and elongation, chromos ...

This chapter reviews some of the many available methods for measuring the binding of myosin and other proteins to actin. Binding to actin has special considerations because actin is a long lattice and the binding site of many of its binding partners consists of multiple actin protomers. The analy ...

Most mammalian proteins undergo reversible protein modification after or during synthesis. These modifications are associated, for the most part, with changes in protein functionality. Protein phosphorylation is the most common posttranslational modification in mammal ...

Cellular microtubules are rigid in comparison to other cytoskeletal elements (1, 2). To facilitate cytoplasmic remodeling and timely responses to cell signaling events, microtubules depolymerize and repolymerize rapidly at their ends (3). These dynamic properties are critica ...

The mouse has become the preferred species for genetic manipulation aimed at creating and studying models for human disease. Although mice are highly resistant to atherosclerosis, dietary induction and, more frequently, gene knockout and transgenic mice have been widely used to study f ...

Heart failure (HF) is a major cause of morbidity and mortality worldwide. Although many therapeutic means are available to prolong the life of HF patients, why HF develops is still poorly understood. Investigators still seek a truly appropriate animal model that will reliably mimic human HF, so ...

Hypertension is an important disease with polygenic inheritance. In order to identify the genes involved in blood pressure regulation, hypertensive rat and mouse models have been developed either by selective breeding or by transgenic technology. The most essential technologic ...

Transgenic and gene-targeted mice are now frequently used to expand the study of cardiac physiology and pathophysiology owing to the ease with which the mouse genome can be manipulated. There are many measures by which an assessment of the phenotypical expression of the transgenic mouse can be ...

The cargo-binding domain of the cytoplasmic dynein complex consists of an intermediate chain, a light-intermediate chain, and three families of light chains. These five subunits form the base of the dynein complex. Variations in the composition and interactions of these subunits play an i ...

Dyneins are highly complex molecular motors containing multiple components that contribute motor, regulatory and cargo-binding activities. Within cilia/flagella, these enzymes comprise the inner and outer arms associated with the doublet microtubules. In this chapter, we d ...