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In Vivo and In Vitro Bioassays for Porcine Ovarian Inhibin and Its Partial Purification
Author(s) -
Yazaki Chiaki,
Hasegawa Yoshihisa,
Miyamoto Kaoru,
Minegishi Takashi,
Yazaki Katsumi,
Igarashi Masao
Publication year - 1982
Publication title -
asia‐oceania journal of obstetrics and gynaecology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.597
H-Index - 50
eISSN - 1447-0756
pISSN - 0389-2328
DOI - 10.1111/j.1447-0756.1982.tb00582.x
Subject(s) - in vivo , bioassay , in vitro , sephadex , follicular fluid , chemistry , medicine , endocrinology , size exclusion chromatography , isoelectric focusing , chromatography , biology , andrology , biochemistry , oocyte , embryo , microbiology and biotechnology , genetics , enzyme
Inhibin activity in porcine follicular fluid (pFF) was quantitatively assayed in both in vivo and in vitro bioassay systems. Post‐castration rise of serum FSH in immature male rats was suppressed dose‐dependently by a single ip injection (0.125 to 1 ml) of steroid‐free pFF. Rat anterior pituitary cell culture was used as an in vitro assay system, in which both release and synthesis of FSH were suppressed dose‐dependently in 0.0125 to 0.4 μ l/ml of steroid‐free pFF. The minimum detectable dose of pFF in vitro was 0.025 μ l/ml and the dose required to produce a half‐maximal suppression (ED 50 ) was 0.06 μ l/ml. Inhibin was partially purified by ammonium sulphate precipitation, DEAE Sephadex ion‐exchange chromatography and Sepharose 6B gel filtration. However, relative specific activity was not increased due to a large loss of inhibin activity. In this respect, isoelectric focusing was very useful for the purification of inhibin.