Neuropeptide Y Protocols: (153 Methods in Molecular Biology)

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.

Table of Contents:
Cloning Techniques.- Cloning Neuropeptide Tyrosine cDNA.- Human Y1/Y5 Receptor Gene Cluster.- Human Type 2 Neuropeptide Y Receptor Gene.- Y4 Receptor in Different Species.- NeuropeptideYY5 Receptor Expression.- Homology-Based Cloning Methods.- Production of Transgenic and Knock-Out Models.- Developing Transgenic NeuropeptideY Rats.- NeuropeptideYY1 Receptor-Deficient Mice.- Antisense Technology.- Antisense Oligonucleotide Approach to Study NPY-Mediated Feeding Signal Transduction.- A G?i RNA-Antisense Expression Strategy to Investigate Coupling of Peptide YY/Neuropeptide Y Receptor to G?i.- NPY Y5 Receptor Subtype.- NPY Antisense Oligodeoxynucleotides to Study the Actions of NPY.- Techniques to Study Tissue Distribution of mRNA.- Localization of Y-Receptor Subtype mRNAs in Rat Brain by Digoxigenin Labeled In Situ Hybridization.- Central Y4 Receptor Distribution.- Combining Non-Isotopic Localization of NPY mRNA with Immunocytochemistry.- Miscellaneous Techniques.- Y Receptors Characterized by RT-PCR.- Quantification of NPY mRNA by Ribonuclease Protection Assay.- Radioligand Binding Studies.

Review :
"...the series is necessary in any library that supports a molecular biology/genetics program...Methods in Molecular Biology is indexed in PubMed, making it very easy to find the protocols. This volume covers neuropeptide Y (NYP), the most abundant peptide in the mammalian nervous system. NYP has also been implicated in the pathophysiology of diseases such as seizures, memory loss, anxiety, and depression, making it an important molecule in clinical research as well. As such, this volume is excellent for any neurobiology or medical collection. The editor of the volume, Ambikaipakan Balasubramaniam, is a well-published researcher in the field"-E-Streams "The editor of this monograph is to be congratulated on bringing together a group of leading experts in the NPY field to describe in a 'user friendly', practical way how to investigate the properties of this peptide....this is an excellent 'cookbook' that will appeal to both the basic and clinical neuroscientist who wishes to study the physiological or pathological effects of this important neuropeptide. The book is nicely produced and the cloth binding gives it the added robustness necessary should it be used at the laboratory bench!"-Human Phychopharmacology "The protocols in this volume cover a wide variety of techniques, from cloning and isolation, to expression and localization. Despite this wide range, each chapter give a quick introduction to the technique and thoroughly discusses the procedures. Methods and materials are specific and clear. A favorite part of all the protocols is the "Notes" section that gives tips and ideas for avoiding some of the usual pitfalls of the technique. Where applicable there are also shortcuts or other ideas to make the protocol work smoothly. The reference section is useful, oftentimes giving early, "historic" references for a technique. There is an extensive index and numerous illustrations, charts and tables to clarify the techniques and results." - E-Streams "This is the first handbook on NPY research techniques and the protocols described in this comprehensive collection can be easily extrapolated to study any peptide hormone, therefore making this volume beneficial to all investigators involved in polypeptide hormone research." - Lavoisier Librairie