title: Development of an anti-vitellin ELISA for the assessment of reproduction in the ridgeback shrimp, Sicyonia ingentis.
abstract: To investigate the reproductive regulation of the ridgeback shrimp, Sicyonia ingentis, vitellin (Vn) synthesis was studied. Using gel filtration chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Vn was found to have a molecular mass of 322 kDa and to be composed of three subunits of 182, 91 and 85 kDa. Purified Vn was used to prepare anti-Vn antiserum, which was used to develop an enzyme-linked immunosorbent assay (ELISA) with a dynamic range of 0.3-300 ng. The ELISA was used to measure hemolymph levels of yolk proteins. The mean hemolymph concentrations in fresh caught animals ranged from 256 (+/-36.6 S.E.M.) to 1073 (+/-87.6 S.E.M.) mg/ml in stage 2 and 4 animals, respectively. The ELISA was also used to determine the effects of steroid hormone injections in adult non-reproductive female shrimp. One milligram injections of progesterone, 17alpha-hydroxyprogesterone or 17beta-estradiol were administered for three consecutive days to individual females. There were no changes in hemolymph vitellogenin levels during the successive 7-day period following the first injection of any steroid.
authors.affiliation: Department of Biology, MS#SB73, California State University, 2555 East San Ramon Avenue, Fresno, CA 93740-8034, USA. briant@csufresno.edu
authors.forename: B
authors.initials: B
authors.lastname: Tsukimura
authors.forename: J S
authors.initials: JS
authors.lastname: Bender
authors.forename: C J
authors.initials: CJ
authors.lastname: Linder
mesh_terms: D019326:17-alpha-Hydroxyprogesterone / Q000008:administration & dosage; D000818:Animals; D002850:Chromatography, Gel; D003429:Cross Reactions; D033364:Decapoda / Q000502:physiology*; D004527:Egg Proteins / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004958:Estradiol / Q000008:administration & dosage; D005260:Female; D008297:Male; D011374:Progesterone / Q000008:administration & dosage; D012098:Reproduction / Q000276:immunology*
chemical_list: D004527:Egg Proteins; D011374:Progesterone; D004958:Estradiol; D019326:17-alpha-Hydroxyprogesterone