RRC ID 3354
Author Kitagawa N, Shiomi K, Imai K, Niimi T, Yaginuma T, Yamashita O.
Title 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.
Journal Zoolog Sci
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.
Volume 22(2)
Pages 213-21
Published 2005-2-1
DOI 10.2108/zsj.22.213
PII JST.JSTAGE/zsj/22.213
PMID 15738641
MeSH Animals Antibodies, Monoclonal / metabolism Bombyx / embryology Bombyx / 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
IF 0.843
Times Cited 23
WOS Category ZOOLOGY
Resource
Silkworms