The ovijector of Ascaris suum: multiple response types revealed by Caenorhabditis elegans FMRFamide-related peptides.

Caenorhabditis elegans possesses 22 FMRFamide-like peptide (flp) genes predicted to encode 60 different FMRFamide-related peptides with a range of C-terminal signatures. Peptides from five flp genes (1, 6, 8, 9 and 14) are known to modulate the ovijector of Ascaris suum in vitro. This study examines the physiological effects of peptides from the remaining 17 flp genes such that the variety of FMRFamide-related peptide-induced ovijector response types can be delineated. Five categories of response were identified according to the pattern of changes in contractile behaviour and baseline tension. Peptides encoded on 16 flp genes (1, 2, 3, 4, 6, 7, 9, 10, 11, 12, 13, 14, 15, 16, 17 and 20) had qualitatively similar inhibitory (response type 1) actions, with the lowest activity thresholds (1 nM) recorded for peptides with FIRFamide or FLRFamide C-terminal signatures. Peptides encoded on four flp genes (2, 18, 19 and 21), and on the A. suum afp-1 gene, had excitatory actions on the ovijector (response type 2), with PGVLRFamides having the lowest activity threshold (1 nM). An flp-2 peptide (LRGEPIRFamide) induced a transient contraction of the ovijector (activity threshold, 10nM) that was designated response type 3. Response type 4 comprised a transient contraction followed by an extended period of inactivity and was observed with peptides encoded on flp-5 (AGAKFIRFamide, APKPKFIRFamide), flp-8 (KNEFIRFamide) and flp-22 (SPSAKWMRFamide). SPSAKWMRFamide was the most potent peptide tested with an activity threshold of 0.1 nM. A single peptide (AMRNALVRFamide; activity threshold 0.1 microM), encoded on flp-11, induced response type 5, a shortening of the ovijector coupled with an increase in contraction frequency. Although most flp genes encode structurally related peptides that trigger one of the five ovijector response types, flp-2 and flp-11 co-encode FMRFamide-related peptides that induce distinct responses. Within the ovijector of A. suum FaRPs play a complex role involving at least five receptor subtypes or signalling pathways.

Physiological effects of FMRFamide-related peptides and classical transmitters on dispersed muscle fibres of the turbellarian, Procerodes littoralis.

The physiological effects of selected classical transmitters and FMRFamide-related peptides (FaRPs) on dispersed muscle fibres from the marine turbellarian, Procerodes littoralis have been examined. Confocal scanning laser microscopy coupled with fluorescein isothiocyanate (FITC) or tetramethylrhodamine (TRITC)-labelled phalloidin revealed a highly developed body wall muscle system with circular, longitudinal and diagonal layers of muscle fibres. Dispersed muscle fibres contracted when depolarized by exposure to extracellular media with elevated K+ (15-100 mM) in a concentration-dependent manner, with a maximal response of 87% achieved at > or = 75 mM. 5-Hydroxytryptamine (5-HT) induced concentration-dependent muscle contraction between 0.01 and 1000 microM, with 10 microM producing a near maximal contraction response of 75%. Acetylcholine (ACh) had less pronounced excitatory effects (0.01-1000 microM), inducing contraction of only 32% of the fibres at 100 microM. The flatworm FMRFamide-related peptides (FaRPs), GYIRFamide, YIRFamide and GNFFRFamide each had concentration-dependent myocontractile effects, indicating the occurrence of at least 1 FaRP receptor on P. littoralis muscle fibres. At 10 microM peptide, GNFFRFamide induced contractions in < 40% of the muscle fibres examined, whereas YIRFamide and GYIRFamide induced contraction in 70 and 75% of muscle fibres, respectively. The order of potency of the peptides was: GYIRFamide > YIRFamide > GNFFRFamide. Pre-incubation of the muscle fibres in 5 microM 5-HT significantly reduced the responses to GYIRFamide, YIRFamide and 5-HT, while the responses to high K+ remained unaltered. Muscle fibres pre-incubated in GYIRFamide (0.1 microM) were also less responsive to 5-HT but not to ACh and high-K+. The GYIRFamide analogue, GYIRDFamide, did not induce muscle contraction (0.01-100 microM) per se, but when co-applied with the myoactive peptides GYIRFamide, YIRFamide or GNFFRFamide, it significantly blocked their ability to elicit contractions. This suggests that the peptides tested may act via a common muscle-based neuropeptide receptor. GYIRDFamide did not alter the contractile effects of high K+, 5-HT or ACh. Collectively, these results indicate that FaRPs, 5-HT and ACh all have the potential to cause muscle contraction in flatworms and that 5-HT and FaRPs alter muscle sensitivity to each other, but do not influence the ability of flatworm muscle fibres to contract.