neuropeptide-f and Monieziasis

The distribution of neuropeptide F (NPF) immunoreactivity in the snail, Helix aspersa, has been demonstrated by immunocytochemistry using 2 region-specific antisera. One, designated NPF3, was raised against a synthetic N-terminal fragment of Helix aspersa NPF; the other, designated PP221, was raised against the C-terminal hexapeptide amide of mammalian pancreatic polypeptide (PP) but cross-reacts fully with the analogous C-terminal region of Helix aspersa NPF. The distribution of NPF immunoreactivity has also been compared with that of FMRFamide using alternate serial sections of Helix aspersa ganglia. Results showed that NPF immunoreactivity was abundant and widespread in the central and peripheral nervous systems and the pattern of immunostaining obtained using both region-specific antisera was similar. Likewise, immunocytochemistry of neural tissues of a congeneric species, Helix pomatia, and 2 prosobranch gastropods, Buccinum undatum and Littorina littorea, produced similar staining patterns with both antisera. However, in the cephalopod mollusc, Loligo vulgaris, and the cestode, Moniezia expansa, positive immunostaining was only obtained with the C-terminal PP antiserum. Immunostaining of alternate serial sections of Helix aspersa ganglia with NPF3, and an antiserum raised to FMRFamide, showed that while a few neurones were immunoreactive with one antiserum only, in the majority, both immunoreactivities were co-localised. NPF thus appears to be an important neuropeptide of widespread distribution in Helix aspersa and the differential immunocytochemical staining obtained using the 2 region-specific antisera would suggest a high degree of primary structural conservation within the gastropod molluscs, but lack of conservation of the N-terminal region of the peptide in other invertebrate groups.

Topics: Amino Acid Sequence; Animals; Cestoda; Decapodiformes; Helix, Snails; Immunohistochemistry; Molecular Sequence Data; Mollusca; Monieziasis; Neuropeptides

Topics: Amino Acid Sequence; Animals; Birds; Cestoda; Conserved Sequence; Helix, Snails; Helminth Proteins; Models, Structural; Molecular Sequence Data; Monieziasis; Neuropeptides; Protein Conformation; Protein Structure, Secondary; Sequence Homology, Amino Acid; X-Ray Diffraction

1. Using immunocytochemical techniques and confocal scanning laser microscopy, the proteocephalidean cestode, Proteocephalus pollanicola from Lough Neagh pollan (Coregomus autumnalis) was examined for the presence of the native platyhelminth neuropeptide, neuropeptide F (NPF). 2. An antiserum specific for whole-molecule NPF (1-39) (Moniezia expansa) did not immunostain nerve processes in P. pollanicola. A C-terminally-directed NPF (30-39) (M. expansa) antiserum immunostained nerve fibres and cell bodies of both the central and peripheral nervous systems, including innervation associated with the female reproductive system. 3. The pattern of immunoreactivity was identical to that obtained using antisera to the C-terminal region of mammalian NPY-superfamily peptides and the invertebrate neuropeptide, FMRFamide. 4. Under radioimmunoassay conditions, only the C-terminally-directed NPF antiserum cross-reacted with the P. pollanicola peptide and detected 58.74 ng/g wet weight of NPF-1 R in extracts of the worm. 5. Chromatographic characterisation of P. pollanicola NPF-immunoreactivity indicated an apparent molecular weight of 4400-4700 Da, similar to that of NPF (M. expansa). 6. Further analytical HPLC characterisation identified two molecular forms of P. pallanicoda NPF-immunoreactivity, both of which had different retention times from those of NPF (M. expansa) and NPF (Artioposthia triangulata). 7. These data suggest that P. pollanicola possesses a neuropeptide which is homologous in its C-terminal region to NPF (M. expansa) but differs in its mid- to N-terminal region.

Topics: Amino Acid Sequence; Animals; Cestoda; Chromatography, Gel; Chromatography, High Pressure Liquid; Fluorescent Antibody Technique; Helminth Proteins; Molecular Sequence Data; Molecular Weight; Monieziasis; Neuropeptides

Immunocytochemical techniques used in conjunction with confocal scanning laser microscopy (CSLM) and electron microscopy have been used to demonstrate, for the first time, the distribution of the parasitic platyhelminth neuropeptide, neuropeptide F (NPF) in the cestode, Moniezia expansa. Antisera were raised to intact NPF(1-39) and to the C-terminal decapeptide of NPF(30-39). These antisera were characterized and validated for use in both immunocytochemistry and radioimmunoassay (RIA). NPF immunoreactivity (IR) was detected using both antisera throughout all of the major components of the central and peripheral nervous systems of the worm. The pattern of NPF-IR was found to mirror the IR obtained using a C-terminally directed pancreatic polypeptide (PP) antiserum and FMRFamide antisera; blocking studies using these antisera revealed that FMRFamide and PP antisera cross-react with NPF(M. expansa). RIA of acid-alcohol extracts of the worm measured 114 ng/g using the C-terminal NPF antiserum and 56 ng/g using the whole-molecule-directed antiserum. While the C-terminally-directed NPF antiserum cross-reacts with NPF-related peptides from other invertebrates, the whole-molecule-directed NPF antiserum is specific for NPF(M. expansa). The C-terminal NPF antiserum has potential for use in the identification and purification of NPF analogues from other platyhelminth parasites.

Topics: Amino Acid Sequence; Animals; Antibodies, Helminth; Antibody Specificity; Cestoda; Chromatography, High Pressure Liquid; Cross Reactions; Helminth Proteins; Immune Sera; Immunohistochemistry; Microscopy, Fluorescence; Microscopy, Immunoelectron; Molecular Sequence Data; Monieziasis; Neuropeptides; Radioimmunoassay; Sheep