Peptidergic neurons, which serve as source of various endocrine neuropeptides, were identified in the suboesophageal ganglion (SG) and brain of insects. In the silkworm Bombyx mori, SG is known to secrete two neuropeptides, diapause hormone (DH) responsible for induction ofembryonic diapause and pheromone biosynthesis-activating neuropeptide, which share a pentapeptide amide, Phe-Xaa-Pro-Arg-Leu-NH2 (Xaa = Gly or Ser), at the C terminus. We have isolated cDNA clones for DH from the cDNA library of SG by using oligonucleotide probes. The molecular characterization of the cDNA reveals that the mRNA encodes an open reading frame consisting of 192 aa residues in which the 24-aa DH peptide is localized at the N-terminal region just after the signal peptide. A homology search proposed that the cDNA encodes pheromone biosynthesis-activating neuropeptide and three other neuropeptides [a-, (3-, and y-SG neuropeptide (SGNP)] in the region following DH, all of which are flanked by possible tryptic cleavage sites and share the Phe-Xaa-Pro-Arg-Leu-Gly sequence at the C terminus. Northern hybridization analysis clearly showed that the gene expression was limited to SG. We chemically synthesized a-, p-, and y-SGNP and used them to identify components in extracts of SG and to examine biological functions. a-and y-SGNP were identified in extracts of SG, and the synthetic P-and y-SGNP expressed weak DH activity.These results indicate that DH, along with four other neuropeptides, is generated from a common precursor polyprotein that is encoded by a single mRNA transcribed in neurosecretory cells of SG.

Abstract:Embryonic diapause of the silkworm, Bombyx mori, is induced by a neuropeptide, diapause hormone (DH), which is secreted from suboesophageal ganglion (SG). cDNA clones for diapause hormone were isolated from a cDNA library constructed from poly A+ RNA of SG. Sequencing from 5'-end to 272 by revealed that the cDNA encodes the DH sequence which is identical to the previously isolated natural peptide except for one amino acid.