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.
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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.
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