Quantitation and Chromatographic Characterisation of Neuropeptide F (NPF) Immunoreactivity in Molluscan Nervous Tissue Using Region-Specific Antisera

Neuropeptide F (NPF) immunoreactivity has been quantified in extracts of the central nervous system from four gastropod (Helix aspersa, Buccinum undatum, Littorina littorea, and Patella vulgaris) and two bivalve (Mytilus edulis and Pecten maximus) molluscs using two region-specific radioimmunoassays. The first employed an antiserum, NPF3, raised to a synthetic N-terminal fragment of H. aspersa NPF and the second employed an antiserum, PP221, raised to the synthetic C-terminal hexapeptide amide of mammalian pancreatic polypeptide which fully cross-reacts with the analogous region of H. aspersa NPF. NPF immunoreactivity was detected in acidified ethanolic brain extracts of the four gastropod molluscs by both antisera. However, only the C-terminally directed antiserum detected immunoreactivity in brain extracts of the two bivalve molluscs. Reverse-phase HPLC analysis of brain extracts from B. undatum and L. littorea resolved a single NPF immunoreactive peptide which was more hydrophobic than natural H. aspersa NPF chromatographed under the same conditions. Gel permeation chromatography of these NPF immunoreactive peptides indicated that they were of a similar molecular mass to that of H. aspersa NPF. These data suggest that NPF is widely distributed in molluscs with a high degree of structural conservation of N- and C-terminal regions within the gastropod molluscs but significant structural differences within the N-terminal regions of analogous peptides in bivalves.