Available primary structural information suggests that the FMRFamide-related peptides (FaRPs) from parasitic and free-living nematodes are different, and that free-living forms may not represent appropriate models for the study of the neurochemistry of parasitic forms in the laboratory. However, here we report the isolation and unequivocal identification of AF2 (originally isolated from the parasite, Ascaris suum) from acidified alcoholic extracts of the free-living species, Panagrellus redivivus. While reverse-phase HPLC analysis of extracts revealed FMRFamide-immunoreactivity to be highly heterogeneous, AF2 was the predominant FMRFamide-immunoreactive peptide present (at least 26 pmol/g wet weight of worms). This peptide was also the major immunoreactant identified by an antiserum raised to the conserved C- terminal hexapeptide amide of mammalian pancreatic polypeptide (PP), which has been used previously to isolate neuropeptide F (NPF). These observations were confirmed by radioimmunoassay and chromatographic fractionation of an acidified alcoholic extract of A. suum heads. The FMRFamide-related peptides present in a nematode extract may be highly dependent on the extraction medium employed, and these data would suggest that this complement of neuropeptides may not be as different between parasitic and free-living nematodes as initial studies have suggested. Finally, all of the evidence suggests that NPF is not present in nematodes and that the PP-immunoreactant previously demonstrated immunochemically is probably AF2.

Peptides belonging to the FMRFamide family are widely distributed amongst invertebrates. We report here on the isolation of the FMRFamide neuropeptide AF2 (Lys-His-Glu-Tyr-Leu-Arg-Phe-NH2) from the parasitic nematode Haemonchus contortus. Immunocytochemical techniques showed that FMRFamide-like material was distributed in several regions of these organisms including nerve cords and cell bodies of the central nervous system. AF2 was isolated using a method that employed 6 steps of reverse-phase HPLC. The concentration of AF2 in this organism was approximately 30 pmol/g of nematode.

AF2 is an endogenous RFamide-like peptide from the parasitic nematode Ascaris suum. The potent stimulatory effects of this peptide on the somatic musculature of Ascaris strongly suggest that it may have an important role in the motornervous system. Here we have investigated the possibility that AF2 may elicit a stimulatory action on Ascaris muscle by potentiating the actions of the excitatory cholinergic motornervous system either pre-synaptically, post-synaptically or both. In in vitro pharmacological experiments AF2 produced a dose-dependent increase in the frequency and amplitude of spontaneous contractions of Ascaris muscle strip which lasted for more than 1 h after a 3 min application of AF2 (10 nM–10µM; N = 7). In addition, AF2 (100 nM) potentiated the contraction elicited by ACh by 43 ± 9% (P < 0·01; N = 8). In electro-physiological recordings from muscle cells, AF2 (10–100 nM; N = 10) potentiated the amplitude of EJPs (excitatory junction potentials). For 100 nM AF2, the potentiation of the EJP was 218 ± 48% (N = 7; P < 0·01). This effect reversed after a wash of 10 min. AF2 did not potentiate the depolarization of the muscle cell elicited by bath applied ACh. These latter two observations are consistent with a presynaptic action of AF2. AF2 (10–100 nM) generated spontaneous muscle cell action potentials in previously quiescent cells. This effect took more than 1 h to wash out. These observations are discussed in terms of the paralysis of Ascaris that is elicited by AF2.