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Antibodies to the microtubule-associated protein tau have been used for more than a decade, both in studies of the role of tau neuronal function, and in examination of the neurofibrillary pathology of Alzheimer’s disease (AD). The vast majority of the available antibodies have been produced w ...

Several lines of evidence suggest a role for membrane ion channels in the neurotoxic effects of the β-amyloid peptide (Aβ). This chapter describes the electrophysiological techniques that can be employed to isolate and record specific membrane conductances that may be altered by Aβ. In gen ...

As noted in the introductory chapters of this book, neuritic plaques composed of accumulated amyloid β (Aβ) peptide are a hallmark pathological feature of the Alzheimer’s disease (AD) brain. Compelling genetic data now implicate these plaques as key causative agents in AD onset, as all known mu ...

The main protein component of the plaques found in the brains of Alzheimer’s disease patients is Aβ, a peptide of 39 to 43 amino acids (reviewed in refs. 1 and 2). Two major Aβ isoforms have been identified in the brains of affected individuals ending at amino acids 40 and 42, respectively (3). The longer form, Aβ42, ...

Until recently, the detergent insolubility of certain membrane-associated proteins was singularly attributed to an association with the cytoskeleton. However, in 1988 we observed that a number of glycosyl-phosphatidylinositol (GPI)-anchored proteins were resistant to s ...

The mammalian inner ear forms from a thickened patch of head ectoderm called the otic placode. The placodal ectoderm invaginates to form a cup whose edges cinch together to establish a fluid-filled sac called the otic vesicle or otocyst. The progenitor cells lining the otocyst lumen will give rise ...

Helios� Gene Gun–mediated transfection is a biolistic method for mechanical delivery of exogenous DNA into cells in vitro or in vivo. The technique is based on bombardment of a targeted cellular surface by micron- or submicron-sized DNA-coated gold particles that are accelerated by a pressu ...

Gene manipulation, specifically in the hair cells of the inner ear during development and adulthood in mice, is crucial for understanding the physiology of hearing and the pathology of deafness in humans. Recent advances have demonstrated that gene expression can be manipulated in devel ...

Greatly impacting the field of neural development are new technologies for generating fate maps in mice and thus for illuminating relationships between embryonic and adult brain structures. Until now, efforts in mammalian models such as the mouse have presented challenges because th ...

The high sensitivity of mammalian hearing is achieved by amplification of the motion of the cochlear partition. The origin of this cochlear amplification is the elongation and contraction of outer hair cells (OHCs) in response to acoustical stimulation. This motility is made possible by a m ...

Regulation of important cellular functions via signaling pathways is a fundamental property of the cell. Intracellular Ca2+ is probably a best known second messenger in cell biology. In mechanosensory cells of the inner ear, the hair cells, intracellular Ca2+ participates in a variety of fu ...

As more and more proteins specific to hair cells are discovered, it becomes imperative to understand their structure and how that contributes to their function. The fluorescence microscopic methods described here can be employed to provide information on protein-protein interact ...

Modern biologists have at their disposal a large array of techniques used to assess the existence and relative or absolute quantity of any molecule of interest in a sample. However, implementing most of these procedures can be a daunting task for the first time, even in a lab with experienced researc ...

Surface plasmon resonance is an optical technique utilized for detecting molecular interactions. Binding of a mobile molecule (analyte) to a molecule immobilized on a thin metal film (ligand) changes the refractive index of the film. The angle of extinction of light, reflected after pola ...

Using gene arrays, it is currently possible to simultaneously measure mRNA levels of many genes in any tissue of interest. Undoubtedly, comprehensive measurements of gene expression as part of carefully designed experiments will continue to further our understanding of audition and ...

Expression of almost every gene is regulated at the transcription level. Therefore, transcriptional factor Transcription factors, consequently, have marked effects on the fate of a cell by establishing the gene expression patterns that determine biological processes. In the aud ...

Structure-function studies of membrane proteins present a unique challenge to researchers due to the numerous technical difficulties associated with their expression, purification and structural characterization. In the absence of structural information, rational id ...

Genomics has provided us with vast amounts of data and thus, the challenge to identify and characterize gene products. Proteomics analysis, using methods such as yeast two-hybrid screenings, isoelectric focusing, and mass spectroscopy, generate potentially useful information. To ...

Functional proteomics comprises a wide range of technologies for the identification of novel protein-protein interactions and biological markers. Studies of protein-protein interactions have gained from the development of techniques and technologies such as immunopre ...

We describe a protocol to screen for protein-protein interactions using the Gal-4 based yeast two-hybrid system. In this protocol, we describe serial transformation of bait into an already constructed cDNA library in yeast AH109 cells. We find this method to gives the most number of true intera ...