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Establishment of a sandwich ELISA system to detect diapause hormone, and developmental profile of hormone levels in egg and subesophageal ganglion of the silkworm, Bombyx mori.

Abstract
In the silkworm Bombyx mori, diapause hormone (DH) is produced in the female subesophageal ganglion (SG) and induces embryonic diapause by targeting developing ovaries. DH is processed from a precursor protein consisting of DH, pheromone biosynthesis activating neuropeptide (PBAN) and three other neuropeptides (SGNPs). Because these five neuropeptides share a common sequence, FXPRLamide, at the C-terminus, a direct and specific assay for DH itself is required in order to understand the profile of concentration changes. In this study, we produced a mouse monoclonal antibody (anti-DH[N] mAb) against the N-terminal region of DH and developed a sandwich enzyme-linked immunosorbent assay using the anti-DH[N] mAb and a rabbit polyclonal antibody against the C-terminus of DH. This procedure enabled us to specifically quantify the DH molecule at femtomolar levels (equivalent to 1/10 of SG). We then plotted DH levels in eggs and SGs during embryonic and post-embryonic development. DH was present in late-stage embryos that had been destined for the production of both diapause and nondiapause eggs. DH levels in SG gradually increased in both types during larval development and peaked at the early pupal stage. At the middle pupal stage, DH levels in SG and SG-brain complex decreased markedly in the diapause-egg producing type, thus indicating active release of DH into the hemolymph. From 5th instar larva to adult, no sexual differences in DH levels were observed in SGs or SG-brain complexes from diapause and nondiapause egg-producing types.
AuthorsNorio Kitagawa, Kunihiro Shiomi, Kunio Imai, Teruyuki Niimi, Toshinobu Yaginuma, Okitsugu Yamashita
JournalZoological science (Zoolog Sci) Vol. 22 Issue 2 Pg. 213-21 (Feb 2005) ISSN: 0289-0003 [Print] Japan
PMID15738641 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Antibodies, Monoclonal
  • Insect Hormones
  • Neuropeptides
  • diapause hormone
Topics
  • Animals
  • Antibodies, Monoclonal (metabolism)
  • Bombyx (embryology, metabolism)
  • Embryo, Nonmammalian (metabolism)
  • Enzyme-Linked Immunosorbent Assay (methods)
  • Female
  • Ganglia, Invertebrate (metabolism)
  • Insect Hormones (metabolism)
  • Larva (metabolism)
  • Male
  • Neuropeptides (isolation & purification)
  • Ovum (metabolism)
  • Sex Factors

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