Neuropeptide Y Protocols

ISBN-13: 9780896036628 / Angielski / Twarda / 2000 / 246 str.

Ambikaipakan Balasubramanian

Opis

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.

Recenzje

"...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

Spis treści

Part I. Cloning Techniques. Cloning Neuropeptide Tyrosine cDNA, Carolyn A. Worby. Human Y1/Y5 Receptor Gene Cluster: Isolation and Characterization, Herbert Herzog. Human Type 2 Neuropeptide Y Receptor Gene: Isolation and Characterization, David A. Ammar and Debra A. Thompson. Y4 Receptor in Different Species: Functional Expression and Binding, Ingrid Lundell, Magnus M. Berglund, and Dan Larhammar. Neuropeptide Y Y5 Receptor Expression, Eric M. Parker. Homology-Based Cloning Methods: Identification of the NPY Y2, Y4, and Y6 Receptors, Douglas J. MacNeil and David H. Weinberg. Part II. Production of Transgenic and Knock-Out Models. Developing Transgenic Neuropeptide Y Rats, Mieczyslaw Michalkiewicz and Teresa Michalkiewicz. Neuropeptide Y Y1 Receptor-Deficient Mice: Generation and Characterization, Thierry Pedrazzini and Josiane Seydoux. Part III. Antisense Technology. Antisense Oligonucleotide Approach to Study NPY-Mediated Feeding Signal Transduction, Sulaiman Sheriff, William T. Chance, and Ambikaipakan Balasubramaniam. A Gai RNA-Antisense Expression Strategy to Investigate Coupling of Peptide YY/Neuropeptide Y Receptor to Gai, Thierry Voisin, Anne-Marie Lorinet, Mathieu Goumain, and Marc Laburthe. NPY Y5 Receptor Subtype: Pharmacological Characterization with Antisense Oligodeoxynucleotide Screening Strategy, Andrea O. Schaffhauser, Alain Stricker-Krongrad, and Karl G. Hofbauer. NPY Antisense Oligodeoxynucleotides to Study the Actions of NPY, Pushpa S. Kalra and Satya P. Kalra. Part IV. Techniques to Study Tissue Distribution of mRNA. Localization of Y-Receptor Subtype mRNAs in Rat Brain by Digoxigenin Labeled In Situ Hybridization, Rachel Parker and Herbert Herzog. Central Y4 Receptor Distribution: Radioactive Ribonucleotide Probe In Situ Hybridization with In Vitro Receptor Autoradiography, Philip J. Larsen and Peter Kristensen. Combining Non-Isotopic Localization of NPY mRNA with Immunocytochemistry, Jennifer Lachey, Jennifer Profitt, Chad Foradori, and RandyJ. Seeley. Part V. Miscellaneous Techniques. Y Receptors Characterized by RT-PCR: Distribution in Rat Intestine, Mathieu Goumain, Thierry Voisin, and Marc Laburthe. Quantification of NPY mRNA by Ribonuclease Protection Assay, Abhiram Sahu. Radioligand Binding Studies: Pharmacological Profiles of Cloned Y-Receptor Subtypes, Karen E. McCrea and Herbert Herzog. Index.

Blurb

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.