Substance P and related peptides in porcine cortex: whole tissue and nuclear localization

A large portion of the knowledge that has been gathered on the distribution of neuropeptides in neural tissues is based on findings obtained with immunocytochemistry and radioimmunoassay. However, these methods give limited structural information about the peptides being studied. Using porcine cortex as a model tissue, we combined immunoaffinity chromatography with reversed-phase high-performance liquid chromatography, radioimmunoassay, and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF-MS). We determined the molecular nature of the peptides contributing to the substance P-like immunoreactivity measured in extracts of whole tissue and cell nuclei. In addition to substance P(1-11), other peptides were extracted using this protocol. The presence of SP(1-11) was confirmed through post-source decay analysis. These results illustrate the usefulness of MALDI-TOF-MS in the characterization of neuropeptides from biological tissues.