The observation that neuropeptide Y (NPY) is the most abundant peptide present in the mammalian nervous system and the finding that it elicits the most powerful orexigenic signal have led to active investigations of the properties of the NPY family of hormones, including peptide YY (PYY) and pancreatic polypeptide (PP). Nearly two decades of research have led to the identification of several NPY receptor subtypes and the development of useful receptor selective ligands. Moreover, these investigations have imp- cated NPY in the pathophysiology of a number of diseases, including feeding disorders, seizures, memory loss, anxiety, depression, and heart failure. Vigorous efforts are therefore continuing, not only to understand the bioche- cal aspects of NPY actions, but also toward developing NPY-based treatments for a variety of disorders. To facilitate these efforts, it was decided to produce the first handbook on NPY research techniques as part of the Methods in Molecular Biology Series. In compiling Neuropeptide Y Protocols, I have gathered contributions on techniques considered critical for the advancement of the NPY field from experts in various disciplines. Each chapter starts with a brief introduction, with Materials and Methods sections following. The latter sections are presented in an easy to follow step-by-step format. The last section of the chapter, Notes, highlights pitfalls and the maneuvers employed to overcome them. This information, not usually disseminated in standard research pub- cations, may prove extremely useful for investigators employing these te- niques in NYP research.
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It is now clear that neuropeptide Y (NPY), the most abundant peptide present in the mammalian nervous system, is implicated in the pathophysiology of many diseases, including eating disorders, seizures, memory loss, anxiety, depression, and cardiovascular diseases, and offers major therapeutic opportunities for their treatment. In Neuropeptide Y Protocols, a strong cohort of established investigators from a variety of disciplines detail their best techniques for advancing NPY research. Among the many readily reproducible methods described are those for cloning, for the production of transgenic and knockout models, and for the use of antisense technology to define the receptors and the signal transduction pathways mediating NPY actions. A range of the major qualitative and quantitative techniques used to study tissue mRNA distribution are also included, as well as methods for radioligand binding. The cloning techniques utilize genomic DNA isolation, expression cloning, PCR cloning, the construction of hybrid receptors, and homology-based cloning.
Highly practical and state-of-the-art, Neuropeptide Y Protocols constitutes a gold-standard collection of proven techniques for advancing today's understanding of this significant neuropeptide, as well as for developing the promising NPY-based therapeutics now emerging for many important disorders.
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