实验仪器使用技术

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 ...

Immunocytochemistry is the localization of a tissue constituent in situ by means of a specific antigen-antibody reaction tagged by a visible label (1). For many years, the technique was not considered sufficiently reliable for the de nova identification of substances, but was used to assess ...

Enzyme-linked immunosorbent assay (ELISA) is a member of the solid-phase immunoassay family that detects specific antigen-antibody binding reactions (1,2). A great many variables of antigen or antibody presentation, treatment, and detection are considered when performing EL ...

The subcellular localization of bioactive peptides, including hormones and neurotransmitters, has immense value, not only in understanding how cells function, but in correlating biochemical and clinical data from tissues in both normal and diseased states. The pioneering immu ...

The radioligand binding assay is a useful tool to characterize peptide receptors and to study the interaction of endogenous and synthetic peptides with those receptors. The most commonly used method is the membrane filtration technique that involves binding of the radioligand to memb ...

The continued modifications in peptide/protein sequencer hardware, derivatization and coupling chemistry, reagent delivery, and component detection provide the protein/peptide chemist with the tools to determine primary structural information on subpicomole quan ...

The blood-brain barrier (BBB) regulates the exchange of substances between the fluids of the central nervous system (CNS) and the blood (1). As such, the BBB is actively involved in providing nutrition and maintaining the homeostatic environment for the brain and spinal cord. Recently, the BBB h ...

It is now well established that the major mechanism for the termination of a neuropeptide signal is not internalization of the peptide, but rather its metabolism by one or more neuropeptidases (reviewed in refs. 1 and 2). By the very nature of their action in hydrolyzing peptides that have been releas ...

The determination of the primary structure of peptides to-date has usually been carried out by automated amino acid sequencing involving Edman chemistry. This sensitive method is reliable, easy to perform, and the interpretation of the results is straightforward. However, other tech ...

Polymerase chain reaction (PCR) techniques comprise some of the most powerful tools in molecular biology research. PCR is characterized primarily by the ability of polymerases to amplify specified regions of DNA both rapidly and efficiently and overcomes the lengthy procedures usu ...

Cells can be defined as “stem cells” when able to self-renew and differentiate into tissue-characteristic cells. Neural stem cells (NSCs) derived from the nervous system are able to generate neuronal and glial cells and are present not only in the developing nervous system, but also in specific r ...

The nervous system is a frequent target of industrial chemicals, pharmaceuticals, and environmental pollutants. To screen large numbers of compounds for their neurotoxic potential, in vitro systems are required which combine organ-specific traits with robustness and high repr ...

Cell cultures are widely used in biomedical research. Primary cultures are directly obtained from fresh tissue and reproduce during days or weeks the major characteristics of the original tissue cells. Primary cell culture systems have shown their usefulness for studying the specific ...

Movement disorders such as Parkinson’s Disease (PD) result, in part, from the selective loss of dopaminergic (DA) neurons. Many studies associated with DA neurodegeneration and neurotoxicity have successfully applied the model organism Caenorhabditis elegans to address PD-re ...

Located at the level of brain capillaries, the blood–brain barrier (BBB) is a crucial component of the neurovascular unit, since its highly regulated properties are needed to maintain optimal conditions for proper neuronal and glial functions. Therefore, understanding BBB features a ...

The choroid plexus epithelium forms the interface between the blood and the cerebrospinal fluid. In addition to its barrier function resulting from the presence of tight junctions sealing the epithelial cells together, the choroid plexus epithelium fulfills vectorial transport ( ...

Recent advances in techniques to introduce nucleic acids into cultured cells have significantly contributed to understanding the roles of genes (and their encoded proteins) in maintaining cellular homeostasis. The objective of this chapter is to provide methodological strate ...

The protein kinase C (PKC) family members include at least 11 different isoforms that, based on their different requirements for activation, have been divided into three subfamilies, the Ca2+-dependent (cPKCα, (βI, βII, and γ), the Ca2+-independent (nPKCδ,ɛ,η,θ, and μ), and phorbol ester-ins ...