Studies on investigation and distribution of the neuropeptides in crustaceans

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Matrix-assisted laser desorption ionization with time-of-flight mass spectrometry (MALDI-TOF MS) has become a rapid and sensitive analytical tool that is having an increasing impact on the direct MS measurement of the secretory granules in the cells. The result is available for the second step that is purification of a given material by monitoring its molecular mass and then sequencing it. The third step is molecular cloning for the precursor of a given substance based on the structural information. The final step is capillary reversed-phase HPLC/Q-Tof analysis with the tissue extract to determine the posttranslational modifications for mature hormone molecules. We have called this total strategy as "topological mass spectrometry analysis". In addition, laser microdissection has become a powerful tool for construction of mass spectrometric morphology in the brain since it is possible to collect single cells from a thin tissue section. In this study, we have identified orcokinin-related peptides, crustacean-SIFamide, and decapod tachykinin-related peptide from the crayfish brain. It is known that the olfactory lobe receives the primary afferent endings of the chemoreceptors on antenna I, but the accessory lobe does not appear to receive a direct input from primary afferent axons. The output pathways from the, olfactory lobe and accessory lobe are provided by the axons of a large population of projection neurons, in which projection neurons innervating the olfactory lobe and accessory lobe innervate the neuropiles of the medulla terminalis and the hemiellipsoid body. The peptides seem to play a significant role in olfaction. Finally, topological mass spectrometry analysis will provide excellent means in micro-characterization of protein/peptide hormone in the field of fish. and insect endocrinology.