Second messengers mediating mechanical responses to the FaRP GYIRFamide in the fluke Fasciola hepatica

Abstract

Spontaneous phasic contractions recorded from isolated body strips of Fasciola hepatica were increased in frequency and amplitude by GYIRFamide, an FMRFamide-related peptide (FaRP). Superfusion with guanosine 5′-O-(2-thiodiphosphate) (100 μM, n = 5) reduced the effects of GYIRFamide on both frequency (by 82%) and amplitude (by 75%). The adenylate cyclase inhibitor MDL-12330A (25 μM) increased spontaneous activity. MDL-12330A completely inhibited the frequency response to GYIRFamide and reduced the amplitude response by 66% as measured relative to this elevated basal activity ( n = 6). Inhibition of phospholipase C (PLC) with neomycin sulfate (1 mM) had no direct effect on activity but reduced the frequency response to GYIRFamide by 64% and the amplitude increase by 95% ( n = 9). The protein kinase C (PKC) inhibitor chelerythrine chloride (10 μM) also reduced frequency and amplitude responses by 98 and 99%, respectively, without affecting basal contractility ( n = 5). Phorbol 12-myristate 13-acetate, an activator of PKC, increased contraction frequency and amplitude ( n = 6). It was concluded that GYIRFamide stimulates mechanical activity in F. hepatica through a G protein, via a PLC- and PKC-dependent second messenger pathway.