Peptide Analysis In Neuronal Cell Culture

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Abstract

Neuropeptides and their analogs often work at extremely low concentrations requiring the development of extremely sensitive techniques for their analysis. To examine the effects of the peptides Vasoactive Intestinal Peptide (VIP) and Pituitary Adenylate Cyclase Activating Peptide (PACAP) on astrocyte cultures, an immunoaffinity capillary electrophoresis (ICE) technique was developed. The procedure involves immobilization of antibodies to the internal surface of a 100-micron diameter fused silica capillary prior to analysis of the conditioned medium derived from the astrocyte cultures. The ICE system was able to isolate and measure 15 different cytokines and chemokines secreted by the stimulated cells. The studies demonstrate that stimulation of astrocytes with pico- to femto-molar concentrations of the neuropeptides induces the production and secretion of a number of inflammatory cytokines and chemokines. These findings indicate a potential role for certain neuropeptides in the local inflammatory process in the central nervous system.