Induction of pheromone production in a moth by topical application of a pseudopeptide mimic of a pheromonotropic neuropeptide.

An amphiphilic analog of Locusta myotropin II (Lom-MT-II), Glu-Gly-Asp-Phe-Thr-Pro-Arg-Leu-amide, was synthesized by addition of 6-phenylhexanoic acid (6-Pha) linked through alanine to the amino terminus. This pseudopeptide, [6-Pha-Ala0]Lom-MT-II, was found to have pheromonotropic activity equivalent to pheromone biosynthesis activating neuropeptide when injected into females of Heliothis virescens. Topical application of [6-Pha-Ala0]Lom-MT-II or Helicoverpa zea-pheromone biosynthesis activating neuropeptide (PBAN), dissolved in dimethyl sulfoxide, to the descaled abdomen of females induced production of pheromone, although more Hez-PBAN than [6-Pha-Ala0]Lom-MT-II was required to obtain significant production of pheromone. Application of [6-Pha-Ala0]Lom-MT-II, dissolved in water, to the abdomen induced production of pheromone, but neither Hez-PBAN nor Lom-MT-II dissolved in water stimulated production of significant amounts of pheromone. Dose- and time-response studies indicated that application of the amphiphilic mimetic in water induced pheromone production in as little as 15 min after application and that the effects were maintained for prolonged periods. These findings show that amphiphilic pseudopeptide mimics of insect neuropeptides will penetrate the insect cuticle when applied topically in water and induce an endogenous response.

Pheromone biosynthesis activating neuropeptides: functions and chemistry.

Sex pheromones are critical for reproductive success in most species of Lepidoptera and their production is regulated by the action of pheromone biosynthesis activating neuropeptides (PBAN). These peptides, composed of 33-34 amino acids, have approximately 80% sequence homology and share the C-terminal sequence FSPRL-NH2, which has been shown to be the minimum sequence required for pheromonotropic activity. This pentamer is structurally similar to the active core (FXPRL-NH2, X = V, T or G) of the insect myotropic pyrokinins. Structure-activity studies have shown that all of the pyrokinins have various degrees of pheromonotropic activity and that some have a superagonistic effect. Peptides that only have sequence homology with PBAN in the C-terminal pentapeptide region, but that are pheromonotropic, also have been identified from months. These findings suggest that induction of pheromone biosynthesis may be regulated by more than one peptide, that PBAN may have a number of physiological functions, and that these peptides regulate induction of pheromone production in a variety of ways.

Potent pheromonotropic/myotropic activity of a carboranyl pseudotetrapeptide analogue of the insect pyrokinin/PBAN neuropeptide family administered via injection or topical application.

A pseudotetrapeptide analogue of the pyrokinin/PBAN or FXPRLamide family (Cbe-Thr-Pro-Agr-Leu-NH2; Cbe = 2-o-carboranylethanoyl-), in which the phenyl ring of the Phe side chain is replaced with the hydrophobic cage-like o-carborane moiety, was synthesized and found to be 10-fold more potent than cockroach leucopyrokinin on an isolated cockroach hindgut bioassay system. In contrast with the naturally occurring peptide, the myostimulatory activity could not be immediately reversed following a saline rinse, providing evidence that the pseudopeptide analogue binds very strongly to the receptor. Once the analogue reaches the receptor, strong receptor binding characteristics may allow it to avoid inactivation by hemolymph peptidases. Although it has an eightfold smaller sequence than the endogenous 33-membered pheromone biosynthesis activating neuropeptide (PBAN), the carboranyl analogue is 10-fold more potent in an in vivo pheromonotropic bioassay of the female tobacco budworm moth Heliothis virescens, demonstrating that the small, C-terminal pentapeptide pyrokinin core analogue contains all the structural information necessary to fully activate pyrokinin receptors. In contrast with PBAN, the amphiphylic carboranyl analogue elicits pheromone production following topical application in aqueous solution to the lateral abdominal surface of H. virescens, providing a noninvasive means of inducing pheromone production in moths. The analogue can potentially serve as a useful tool to insect researchers studying, and/or attempting to disrupt, physiological processes regulated by pyrokinin-like neuropeptides in insects. A possible role for this and related pyrokinin analogues in future pest insect management strategies is briefly discussed.

Pheromonotropic activity of naturally occurring pyrokinin insect neuropeptides (FXPRLamide) in Helicoverpa zea.

Insect neuropeptides, having the common C-terminal sequence FXPRLamide X = V, T, S, or G), were tested for phyeromonotropic activity in the moth, Helicoverpa zea. Dose-response studies indicated that locustamyotropin-II or locustapyrokinin-II induced production of more pheromone than was stimulated by the pheromone biosynthesis activating neuropeptide of this moth. Other peptides showed various degrees of pheromonotropic activity. The data indicated that substitution of the variable amino acid in the C-terminal pentapeptide sequence resulted in significant differences in pheromonotropic activity. However, the overall structure of the peptide was also found to be of importance.