The development and analysis of transgenic mice that overexpress or misexpress neurotrophins or their receptors has led to a number of fundamental insights into the mechanisms whereby this family of growth factors exerts its biological effects in vivo (1–5). These transgenic mice were g ...

The ability to manipulate the mouse genome has provided extremely useful model systems for studying the underlying molecular mechanisms of neurotrophin peptide action in mammalian development and adult systems. One way to use transgenics is to overexpress neurotrophins in neuro ...

The development of a simple and “gentle” technique for the radio-iodination of proteins with lactoperoxidase (1) and subsequent modifications of this technique (2–6) have made it possible to introduce labeled neurotrophins as sensitive tracers into living organisms and to determi ...

Neurotrophic factors play an essential role in the growth, survival, and differentiation of neurons in the nervous system. Several well-characterized neurotrophic factors, such as nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), NT-4 ...

Selective destruction of neurons based on the use of targeted toxins has proven successful for several types of neurons (1). This chapter will describe the use of an immunotoxin to selectively destroy rat neurons that express the low-affinity neurotrophin receptor (p75NTR) (2). This immun ...

Duplex formation between an oligonucleotide and a strand of mRNA can effectively inhibit the expression of a specific gene via interfering with the cellular protein synthesis process. The antisense technology (1) is regarded as a powerful tool in molecular biology. Advances in the field h ...

In order to look at the retrograde transport of neurotrophic factors from the target tissue to the innervating cell body, the neurotrophin needs to be labeled to a high specific activity. This chapter outlines methods used to bioassay the labeled neurotrophins and the advantages and disadva ...

In recent years, we have witnessed an explosion in the number of techniques available for selectively perturbing the genes and molecules that regulate tissue growth and maintenance. From drugs and neutralizing antibodies, we have progressed to gene deletion, “dominant negative,” ant ...

Polyacrylamide gel electrophoresis (PAGE) in buffers containing the anionic detergent sodium dodecylsulfate (SDS) is a very powerful technique for small-scale separation of polypeptides and for assigning molecular weights to these molecules. However, the majority of syste ...

In today’s world, we face a significant degree of miniaturization in many aspects of technology. Wonders of miniaturization are also occurring in the areas of physical and biological sciences. The development of a 21st-century technology has enabled investigations and processes to be p ...

Fast atom bombardment (FAB) was introduced as a new ionization technique (1) in 1981 by M. Barber and his coworkers. This was a breakthrough in the analysis of unstable and involatile compounds such as peptides, which were difficult to study by other ionization methods. FAB employs a particle beam co ...

Atmospheric pressure ionization (API) interfaces have become powerful and popular tools for sample ionization (so-called soft ionization) and are primarily used in the analysis of polar and thermolabile compounds (e.g., peptides, proteins). Detailed descriptions of the most imp ...

This method describes how, by mixing peptides with a UV-absorbing matrix, their masses can be determined using matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS).

Circular dichroism (CD) spectroscopy is currently the method of choice to study the conformation of proteins in low resolution (1). Chiral substances absorb right and left circularly polarized lights to different extents, demonstrating differences in absorbance (△A) and molar exti ...

1H nuclear magnetic resonance (NMR) has proven to be a uniquely powerful tool for studying the structure of peptides in solution. I will concentrate on the type of structural information that is obtainable from NMR, the types of spectra needed to get this information, and how to interpret these spec ...

An X-ray crystal structure of a neuropeptide bound to its native receptor protein would provide a wealth of information concerning the structural requirements for ligand-receptor binding. Unfortunately, many neuropeptide-protein complexes are very difficult, if not impossi ...

Molecular modeling is the science of the generation, manipulation, and representation of three-dimensional structures of molecules using computational chemistry and high resolution computer graphics. Since peptides of biological interest are large molecules, molecul ...

Neuropeptides vary appreciably in terms of then molecular mass, charge, and hydrophobicity so that there is no single optimum method for their extraction from biological materials such as tissues, cultured neurons, plasma, or cerebrospinal fluid (CSF). As all neuropeptides are rapid ...

Since an immunogen requires both an antigenic site and a T-cell receptor binding site, there is a minimum size necessary (1). Natural immunogens have a molecular weight 5000. Small molecules such as neuropeptides may be able to bind to the surface of B-cells, but do not stimulate an immune response. Such ...

There is frequently a need to measure concentrations of neuropeptides in tissue perfusates, tissue extracts, chromatographic column fractions, and so on. Since the concentrations of neuropeptides encountered are often low (usually the low fmol/mL range in perfusates and pmol/g ran ...