Identification, characterization and immunological localization of a novel myotropic neuropeptide in the Colorado potato beetle, Leptinotarsa decemlineata

A novel myotropic heptapeptide was isolated from an extract of 54,000 heads of adult Leptinotarsa decemlineata by means of high performance liquid chromatography (HPLC), using the Locusta migratoria oviduct motility bioassay as monitoring system. The full primary structure was established as H-Ala-Tyr-Asn-Gly-Pro-Leu-Ala-NH2. This peptide, designated as Led-MNP-I, has a unique structure and does not belong to any known vertebrate or invertebrate peptide family. Two adjacent Led-MNP-I-immunoreactive perikarya were found in each optic lobe and in each half of all thoracic ganglia. Its absence from the pars intercerebralis and neurohemal organs suggests that Led-MNP-I is not a neurohormone but a neurotransmitter or neuromodulator. Treatment of isolated oviducts with varying concentrations of Led-MNP-I did not elicit significant changes in the level of cAMP concentration, suggesting that cAMP does not act as a second messenger for Led-MNP-I. Instead, Led-MNP-I induces an elevation of IP3. Treatment with Led-MNP-I did not stimulate cAMP production in the Colorado beetle brain, but this could be due to the very small number of receptive cells present. Both tissues contained a forskolin-sensitive adenylate cyclase enzyme.status: publishe