Inhibin production by porcine granulosa and luteal cells: development and biological validation of a RIA.
We describe the development and biological validation of a radioimmunoassay for immuno- and bioactive porcine inhibin. A synthetic 1-32 porcine inhibin peptide was used to raise an antiserum and Tyr-1-32 peptide as tracer. As standard we employed porcine follicular fluid calibrated with the 1-32 alpha-inhibin. Medium obtained from serum-free cultured porcine granulosa cells was chromatographed on Superose S-12 and Mono-Q. Resulting fractions were analysed for inhibin bio- and immunoreactivity. It is shown that granulosa cells produce at least two types of bioactive inhibins, one being also immunoactive in our RIA. We studied secretion of immunoreactive inhibin from porcine ovarian cells under various conditions: Inhibin secretion from mature and immature granulosa cells can be stimulated by FSH, whereas hCG enhances inhibin secretion only from mature granulosa cells. During extended time of culture, the capability of granulosa cells to secrete inhibin is reduced. In contrast, progesterone secretion from these cells increases; this is due to spontaneous functional luteinization. This assumption is supported by the low inhibin secretion of luteal cells in comparison to granulosa cells. Intracellular inhibin content in luteal cells is below detection limit of the RIA, whereas granulosa cells contain readily detectable amounts of this hormone.
Michel
U
U
Department of Obstetrics/Gynecology, University of Göttingen.
Jarry
H
H
Metten
M
M
Wuttke
W
W
D000818:Animals; D002478:Cells, Cultured; D003338:Corpus Luteum / Q000166:cytology / Q000378:metabolism*; D005260:Female; D006107:Granulosa Cells / Q000378:metabolism*; D007265:Inhibins / Q000096:biosynthesis*; D010902:Pituitary Gland / Q000166:cytology / Q000378:metabolism; D011863:Radioimmunoassay / Q000379:methods*; D013552:Swine
D007265:Inhibins