Neuropeptide F (Moniezia expansa): localization and characterization using specific antisera.
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.
Maule
A G
AG
Comparative Neuroendocrinology Research Group, School of Biology and Biochemistry, Queen's University of Belfast, U.K.
Shaw
C
C
Halton
D W
DW
Brennan
G P
GP
Johnston
C F
CF
Moore
S
S
D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000096:biosynthesis / Q000276:immunology; D000918:Antibody Specificity; D002589:Cestoda / Q000737:chemistry* / Q000276:immunology / Q000648:ultrastructure; D002851:Chromatography, High Pressure Liquid; D003429:Cross Reactions; D015801:Helminth Proteins / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D007106:Immune Sera / Q000276:immunology; D007150:Immunohistochemistry; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D008989:Monieziasis / Q000469:parasitology; D009479:Neuropeptides / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D011863:Radioimmunoassay; D012756:Sheep
D000909:Antibodies, Helminth; D015801:Helminth Proteins; D007106:Immune Sera; D009479:Neuropeptides; C076101:neuropeptide F