Neurotrophin Protocols

ISBN-13: 9781489939838 / Angielski / Miękka / 2013 / 274 str.

Robert A. Rush

Opis

The past decade has seen an extraordinary growth in research interest in neurotrophic factors, and the study of the neurotrophin family has led this activity. Nevertheless, this area of research has often struggled as a result of techniques that were either inadequate or just emerging from other research fields and disciplines. Neurotrophin Protocols has brought together many leaders in the neurotrophin field who detail their special expertise in a wide variety of techniques. Though most procedures are valid across many diff- ent fields of research, some of those described here have been developed to address particular issues within the neurotrophic factor field. The protocols cover a broad range of biochemical, histological, and biological techniques that are often required by the modern laboratory. However, all have been written with sufficient detail to allow any laboratory to achieve proficiency without need of reference to other texts. Neurotrophin Protocols is divided into four sections dealing with p- tein, RNA, recombinant, and in vivo techniques. Protein techniques have in general been less successfully employed than those dealing with RNA or DNA. However, procedures that achieve localization and quantification of the neurotrophins are now being used more extensively. Their inclusion here should assist further studies at the protein level. Transgenic cell lines and animals are commonplace in the scientific research literature, but their inc- sion in several chapters in this book provide some novel uses that are not readily available elsewhere.

Recenzje

"...provides a collection of chapters written by experts in developmental neuroscience, neuroanatomy, signaling mechanisms that regulate neuronal survival and plasticity, and neurodegenerative disorders....This book will be a valuable addition to libraries at Universities and various other institutes engaged in biomedical sciences, as well as to the personal collection of those scientists interested in growth factors....A strength of Neurotrophin Protocols is that it covers considerable ground with little repetition of background information and specific methods among chapters....we expect that this book will spend less time on the shelf and more time on lab benches and office desks. This is well done and timely contribution to the field of neurotrophic factor research."-Journal of Molecular Neuroscience

Spis treści

Part 1. Neurotrophin Quantification and Localization. Localization of Neurotrophin Proteins Within the Central Nervous System by Immmunohistochemistry, James M. Conner. Neurotrophin Immunohistochemistry in Peripheral Tissues, Robert A. Rush and Xin-Fu Zhou. Extraction and Quantification of the Neurotrophins, Shu-Hua Zhang, Xin-Fu Zhou, and Robert A. Rush. Identifying Novel Proteins in Nervous Tissue Using Microsequencing Techniques, Selena E. Bartlett. Part 2. RNA Quantification and Localization. Quantification of mRNA Levels Using Northern Blotting, Paul Fernyhough. Quantification of mRNA Levels Using Ribonuclease Protection Assay, Pirkko Henttu. Quantification of Neurotrophin mRNA by RT-PCR, Edward Chie, Dennis Liu, Xin-Fu Zhou, and Robert A. Rush. Sensitive and Nonradioactive In Situ Detection of Neurotrophin mRNAs in the Nervous System, Cory J. Xian and Xin-Fu Zhou. Part 3. Construction of Recombinant Proteins, and Transgenic Cells and Animals. The Expression of Neurotrophic Factors such as GDNF in the E. coli System, Bing-Ren Huang and Xue-Mei Ma. Construction of Cells Expressing Neurotrophins, Jasodhara Ray and Fred H. Gage. Efficient Generation of Stable Pheochromocytoma (PC12) Cell Lines Using a Recombinant Retrovirus (LNC), Matthew E. Cunningham, Jan K. Kitajewski, and Lloyd A. Greene. Construction and Analysis of Transgenic Animals Overexpressing Neurotrophins, Kathryn M. Albers and Brian M. Davis. Construction of Transgenic Animals Overproducing Neurotrophins and Their Receptors, Carrie G. Causing and Freda D. Miller. Part 4. In Vivo Analysis. Tracing with Radiolabeled Neurotrophins, Christopher S. von Bartheld. Toxin-Induced Death of Neurotrophin-Sensitive Neurons, Ronald G. Wiley. Oligonucleotides as Inhibitors of Protein Synthesis, W. Ruwan Epa, Graham L. Barrett, and Perry F. Bartlett. The IODO-GEN Method for Labeling and the Use of Retrograde Axonal Transport to Assay Neurotrophins, Ian A. Hendry and Anna J. Reynolds. Stereological Estimation of Neuronal Number: The Optical Disector/Cavalieri Combination, Victor Nurcombe, Nigel G. Wreford, and John F. Bertram. Index.

Blurb

Despite the extraordinary growth of research interest in neurotrophic factors, the techniques available have often been inadequate or just emergent from other disciplines. In Neurotrophin Protocols, established leaders in the neurotrophin field detail their special expertise in a wide variety of key protein, RNA, recombinant, and in vivo techniques. The protocols range from immunological analysis for the cellular localization and quantification of the neurotrophins, to genetic manipulation of cells and animals for the analysis of biological function, to quantitative analysis of the active neurotrophin genes. There are also radiotracing techniques for studying neurotrophin transport in both the retrograde and anterograde directions, procedures for using immunotoxins to study the effects of eliminating a single class of neurons, and the essential stereological method for estimation of neuronal numbers. Each method includes not only detailed step-by-step instructions, but also a list of necessary equipment and supplies, and valuable notes spelling out quick tips and tricks of the trade.
Timely and robust, Neurotrophin Protocols provides today's neuroscientists in both academia and industry with a comprehensive range of practical, readily reproducible methods for studying neurotrophins and the critically important effects they have on the nervous system.