Abstract
Purpose
Immunoreactive inhibin (i-inhibin) has been reported to be present in the peritoneal fluid of women. The radioimmunoassay employed measures free, biologically inactive α-subunits(s) equally as well as dimeric, biologically active inhibin. The present study was designed to determine if biologically active, dimeric inhibin is present in the peritoneal fluid of women.
Methods
Peritoneal fluid of four women was assayed by radioimmunoassay, a sheep pituitary bioassay, and two ELISA procedures which utilized specific monoclonal antibodies for the “capture” of the α-subunit (ELISA-A) or the β-subunit (ELISA-B) of inhibin and subsequent quantification of dimeric inhibin-A.
Results
There was a good correlation between the values obtained by radioimmunoassay, bioassay, and both ELISAs; two samples (from the late follicular phase) with relatively high i-inhibin concentrations were positive in all four assays, whereas two samples (from the early follicular phase) with very low i-inhibin concentrations were negative in the bioassay and ELISAs.
Conclusion
A significant portion of the immunoreactive inhibin in the peritoneal fluid obtained during the late follicular phase of women is dimeric, biologically active inhibin. We speculate that this may have potential implications for oocyte maturation and early embryogenesis within the oviduct.
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References
Hemmings R, Falcone T, Billiar RB, Tulandi T: Peritoneal fluid inhibin during the menstrual cycle. Obstet Gynecol 1992;80:27–29.
McLachlan RI, Robertson DM, Burger HG, de Kretser DM: The radioimmuno-assay of bovine and human follicular fluid and serum inhibin. Mol Cell Endocrinol 1986;46:175–185.
O W-S, Robertson DM, de Kretser DM: Inhibin as an oocyte meiotic inhibitor. Mol Cell Endocrinol 1989;62:307–311.
Alak BM, Woodruff TK, Stouffer RL, Wolf DP: In vitro enhancement of primate oocyte maturation and fertilization by inhibin A and activin A. 1993; Am Fertil Soc Meeting Abstract 0-083.
Phocas I, Sarandakou A, Rizos D, Dimitriadou F, Mantzavinos T, Zourlas PA: Secretion of α-immunoreactive inhibin by human preembryos cultured in vitro. Hum Reprod 1992;7:545–549.
Roberts VJ, Sawchenko PE, Vale W: Expression of inhibin/activin subunit messenger ribonucleic acids during rat embryogenesis. Endocrinology 1991;128:3122–3129.
Schneyer AL, Mason AJ, Burton LE, Ziegner JR, Crowley WF Jr. Immuno-reactive inhibin α-subunit in human serum: implications for radioimmunoassay. J Clin Endocrinol Metab 1990;70:1208–1212.
Knight PG, Beard AJ, Wrathall JHM, Castillo RJ: Evidence that the bovine ovary secretes large amounts of monomeric inhibin alpha subunit and its isolation from bovine follicular fluid. J Mol Endocrinol 1989;2:189–200.
Robertson DM, Giacometti M, Foulds LM, Lahnstein J, Goss NH, Hearn MTW, deKretser DM: Isolation of inhibin α-subunit precursor proteins from bovine follicular fluid. Endocrinology 1989;125:2141–2149.
Sugino K, Nakamura T, Takio K, Titani K, Miyamoto K, Hasegawa Y, Igarashi M, Sugino H: Inhibin alpha-subunit monomer is present in bovine follicular fluid. Biochem Biophys Res Commun 1989;159:1323–1329.
Miyamato K, Hasegawa Y, Fukuda M, Igarashi M: Demonstration of high molecular weight forms of inhibin in bovine follicular fluid (bFF) by using monoclonal antibodies to (bFF) 32K inhibin. Biochem Biophys Res Comm 1986;136:1103–1109.
Robertson DM, Foulds LM, de Vos F, Leversha L, de Kretser DM: Identification of inhibin and inhibin-related proteins in human follicular fluid. Reprod Fertil Dev 1990;2:327–335.
Sugino K, Nakamura T, Takio K, Miyamoto K, Hasegawa, Y, Igarashi M, Titani K, Sugino H: Purification and characterization of high molecular weight forms of inhibin from bovine follicular fluid. Endocrinology 1992;130:789–796.
Betteridge A, Craven RP: A two-site enzyme-linked immunosorbent assay for inhibin. Biol Reprod 1991;45:748–754.
Gaines Das RE, Rose M, Zanelli JM: International collaborative study byin vitro bioassays of the first International Standard for porcine inhibin. J Reprod Fertil 1992;96:803–814.
Billiar RB, Richardson DW, Little B: Reduced serum inhibin concentrations during ovulatory cycles of estrogen-treated rhesus monkeys: an indicator of FSH bioactivity. Endocrinology 1991;128:2280–2284.
Tsonis CG, McNeilly AS, Baird DT: Measurement of exogenous and endogenous inhibin in sheep serum using a new and extremely sensitive bioassay for inhibin based on inhibition of ovine pituitary FSH secretionin vitro. J Endocrinol 1986;110:341–352.
Groome N: Ultrasensitive two-site assays for inhibin-A and activin-A using monoclonal antibodies raised to synthetic peptides. J Immunol Meth 1991;145:65–69.
Groome N, O'Brien M: Immunoassay for inhibin and its subunits. Further application of the synthetic peptide approach. J Immunol Meth 1993;165:167–176.
Groome N, Illingworth PJ, O'Brien M, Cooke I, Ganesan TS, Baird DT, McNeilly AS: Detection of dimeric inhibin throughout the human menstrual cycle by two-site enzyme immunoassays. Clin Endocrinol 1994;40:717–723.
Krummen LA, Woodruff TK, De Guzman G, Cox ET, Baly DL, Mann E, Garg S, Wong W-L, Cossum P, Mather JP: Identification and characterization of binding proteins for inhibin and activin in human serum and follicular fluids. Endocrinology 1993;132:431–443.
Bramley TA, Menzies GS, Baxter G, Webb R, McNeilly AS: Apparent α-inhibin subunit immunoactivity in porcine and ovine luteal extracts is due to interference by cytosolic proteases in the assay. J Endocrinol 1992;134:341–352.
Groome N, Hancock J, Betteridge A, Lawrence M, Craven R: Monoclonal and polyclonal antibodies reactive with the 1–32 amino terminal sequence of the alpha subunit of human 32K inhibin. Hybridoma 1990;9:31–41.
Tierney ML, Goss NH, Tomkins SM, Kerr DB, Pitt DE, Forage RG, Robertson DM, Hearn MTW, de Kretser DM: Physicochemical and biological characterization of recombinant human inhibin A. Endocrinology 1990;126:3268–3270.
Roberts VJ, Barth S, El-Roeiy A, Yen SSC: Expression of inhibin/activin subunits and follistatin messenger ribonucleic acids and proteins in ovarian follicles and corpus luteum during the human menstrual cycle. J Clin Endocrinol Metab 1993;77:1402–1410.
Yamoto M, Minami S, Nakano R, Kobayashi M: Immunohistochemical localization of inhibin/activin subunits in human ovarian follicle during the menstrual cycle. J Clin Endocrinol Metab 1992;74:989–993.
Groome N, Lawrence M: Preparation of monoclonal antibodies reactive with the β-A subunit of human ovarian inhibin. Hybridoma 1991;10:309–316.
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Billiar, R.B., Hemmings, R., Smith, P. et al. Identification of biologically active inhibin in the peritoneal fluid of women. J Assist Reprod Genet 12, 55–60 (1995). https://doi.org/10.1007/BF02214130
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DOI: https://doi.org/10.1007/BF02214130