Defining the Neuropeptidome of the Spiny Lobster Panulirus interruptus Brain Using a Multidimensional Mass Spectrometry-Based Platform

摘要: Decapod crustaceans are important animal models for neurobiologists due to their relatively simple nervous systems with well-defined neural circuits and extensive neuromodulation by a diverse set of signaling peptides. However, biochemical characterization these endogenous neuropeptides is often challenging limited sequence information about neuropeptide genes the encoded preprohormones. By taking advantage homology in observed related species using home-built crustacean database, we developed semi-automated sequencing strategy characterize neuropeptidome Panulirus interruptus, an aquaculture species, few known preprohormone sequences. Our streamlined process searched high mass accuracy high-resolution data acquired on LTQ-Orbitrap flexible algorithm ProSight that allows discrepancy from reported sequences our resulting detection 32 neuropeptides, including 19 novel ones. We further improved overall coverage 51 multidimensional platform employed multiple analytical techniques dimethylation-assisted fragmentation, de novo nanoliquid chromatography-electrospray ionization-quadrupole-time-of-flight (nanoLC-ESI-Q-TOF), direct tissue analysis, spectrometry imaging matrix-assisted laser desorption/ionization (MALDI)-TOF/TOF. The discovery rate this unsequenced model organism demonstrated utility pipeline highlighted utilizing strategies. Collectively, study expands catalog more importantly presents approach can be adapted exploring possess information.