Characterizing the Hez-PBAN gene products in neuronal clusters with immunocytochemistry and MALDI MS

Methods to characterize pheromone biosynthesis activating neuropeptide (PBA N) and other PBAN gene encoded neuropeptides (PGN) from individual subesoph ageal ganglion neuronal clusters of the corn earworm moth, Helicoverpa tea, were developed. Individual antisera against the N-terminal sequence to PBA N and each of the three PGNs from the Hez-PBAN prohormone were developed, a nd their specificity determined. In all cases, each antiserum stains the sa me three groups of subesophageal ganglion ventral midline neurons--the mand ibular, maxillary and labial neurons---in both adult females and males. The se results were confirmed using matrix assisted laser desorption/ionization mass spectrometry (MALDI MS) of individual subesophageal ganglion neuronal clusters. Using mass spectrometry, the amidated PGN-24 was not detected bu t an N-terminally extended form is observed that is two amino acids longer. Other peptides resulting from the processing of the Hez-PBAN prohormone we re detected. Using both the specific antisera and the cellular profiling ab ilities of MALDI MS, the roles of individual members of the Hez-PBAN prohor mone derived peptides can now be explored. (C) 2000 Elsevier Science Ltd. A ll rights reserved.