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Radioimmunoassay of relaxin-like gonad-stimulating peptide in the starfish Patiria (=Asterina) pectinifera.
Yamamoto, Kazutoshi; Kiyomoto, Masato; Katayama, Hidekazu; Mita, Masatoshi.
Afiliación
  • Yamamoto K; Department of Biology, Faculty of Education and Integrated Sciences, Center for Advanced Biomedical Sciences, Waseda University, Wakamatsucho 2-2, Shinjuku-ku, Tokyo 162-8480, Japan.
  • Kiyomoto M; Tateyama Marine Laboratory, Marine and Coastal Research Center, Ochanomizu University, Kou-yatsu 11, Tateyama, Chiba 294-0301, Japan.
  • Katayama H; Department of Applied Biochemistry, School of Engineering, Tokai University, Kitakaname 4-1-1, Hiratsuka, Kanagawa 259-1292, Japan.
  • Mita M; Department of Biology, Faculty of Education, Tokyo Gakugei University, Nukuikita-machi 4-1-1, Koganei-shi, Tokyo 184-8501, Japan. Electronic address: bio-mita@u-gakugei.ac.jp.
Gen Comp Endocrinol ; 243: 84-88, 2017 03 01.
Article en En | MEDLINE | ID: mdl-27838378
A relaxin-like gonad-stimulating peptide (RGP) from starfish Patiria (=Asterina) pectinifera is the first identified invertebrate gonadotropin for final gamete maturation. An antiserum against P. pectinifera RGP (PpeRGP) was produced by immunizing rabbits with a PpeRGP sulfanyl-polyethylene glycol derivative conjugated with keyhole limpet hemocyanin (KLH) as the antigen. The antiserum was used for the development of a specific and sensitive radioimmunoassay (RIA) for the measurement of RGP. In binding experiments using radioiodinated PpeRGP and antiserum against PpeRGP, a displacement curve was obtained using radioinert PpeRGP. The sensitivity of the RIA, defined as the amount of PpeRGP that significantly decreased the counts by 2 SD from the 100% bound point, averaged 0.040±0.002pmol PpeRGP per 100µl assay buffer (0.40±0.02nM) in 10 assays. Intra-assay and inter-assay coefficients of variation were 6.1% and 2.7%, respectively. Serial dilution of whole homogenates from the radial nerve cords and circumoral nerve-rings of P. pectinifera produced displacement curves parallel to the PpeRGP standard. Thus, the amounts of PpeRGP were determined as 1.54±0.09pmol/mg wet weight of radial nerves and 0.87±0.04pmol/mg wet weight of nerve-rings, respectively. On contrary, pyloric stomach, pyloric caeca, tube-feet, ovaries, testes, and ovarian follicle cells did not react in the RIA system. Furthermore, the A- and B-chains of PpeRGP, Asterias amurensis RGP, bovine insulin, and human relaxin did not show cross-reactivity in the RIA. These results strongly suggest that the RIA system is a highly specific and sensitive with respect to PpeRGP.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Fragmentos de Péptidos / Relaxina / Radioinmunoensayo / Asterina / Gónadas / Hormonas de Invertebrados Límite: Animals Idioma: En Revista: Gen Comp Endocrinol Año: 2017 Tipo del documento: Article País de afiliación: Japón Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Fragmentos de Péptidos / Relaxina / Radioinmunoensayo / Asterina / Gónadas / Hormonas de Invertebrados Límite: Animals Idioma: En Revista: Gen Comp Endocrinol Año: 2017 Tipo del documento: Article País de afiliación: Japón Pais de publicación: Estados Unidos