Neuropeptide precursor processing detected by triple immunolabeling.
Peptides that play critical physiological roles are often encoded in precursors that contain several gene products. Differential processing of a polypeptide precursor by cell-specific proteolytic enzymes can yield multiple messengers with diverse distributions and functions. We have isolated SDNFMRFamide, DPKQDFMRFamide, and TPAEDFMRFamide from Drosophila melanogaster. The peptides are encoded in the FMRFamide gene and have a common C-terminal FMRFamide but different N-terminal extensions. In order to investigate the regulation of expression of FMRFamide peptides, we generated antisera to distinguish between the structurally related neuropeptides. We established a triple-label immunofluorescence protocol using antisera raised in the same host species and mapped the neural distribution of SDNFMRFamide, DPKQDFMRFamide, and TPAEDFMRFamide. Each peptide has a unique, nonoverlapping cellular expression pattern, suggesting that the precursor is differentially processed. Thus, our data indicate that D. melanogaster contains cell-specific proteolytic enzymes to cleave a polypeptide protein precursor, resulting in unique expression patterns of neuropeptides.
McCormick
J
J
Biology Department, University of Michigan, Ann Arbor 48109-1048, USA.
Lim
I
I
Nichols
R
R
D000818:Animals; D004330:Drosophila / Q000378:metabolism; D019835:FMRFamide / Q000302:isolation & purification*; D005455:Fluorescent Antibody Technique; D019476:Insect Proteins / Q000378:metabolism*; D009479:Neuropeptides / Q000378:metabolism*; D018377:Neurotransmitter Agents / Q000302:isolation & purification; D011498:Protein Precursors / Q000378:metabolism*; D011499:Protein Processing, Post-Translational
D019476:Insect Proteins; D009479:Neuropeptides; D018377:Neurotransmitter Agents; D011498:Protein Precursors; C098192:SDNFMRFamide; C121685:aspartyl-prolyl-lysyl-glutaminyl-aspartyl-phenylalanyl-methyonyl-arginyl-phenylalanylamide; C121688:threonyl-prolyl-alanyl-glutamyl-aspartyl-phenylalnyl-methionyl-arginyl-phenylalanylamide; D019835:FMRFamide