Z Blumenfeld1, M Ritter. 1. Department of Obstetrics and Gynecology, Rambam Medical Center and the B. Rappaport Faculty of Medicine, Technion, Israel Institute of Technology, Haifa. bzeev@techunix.technion.ac.il
Abstract
BACKGROUND: Activin has been previously demonstrated to directly stimulate the synthesis of gonadotropin-releasing hormone (GnRH) receptors and to increase follicle-stimulating hormone (FSH) secretion in nonhuman pituitary cell cultures (PCCs). Currently, knowledge of the physiological role of these peptides in primates is still far from complete. Moreover, several results in macaque monkeys failed to support an unequivocal role for inhibin in FSH suppression. Whereas the bioactivity of inhibin and activin has been demonstrated in rat PCCs, no data exist on human pituitary response to these peptides either in vivo or in vitro. METHODS: We studied the secretion of FSH and luteinizing hormone (LH) by dispersed human fetal pituitary cells from midtrimester abortions in response to recombinant human (rh-) activin-A, inhibin-A, and other secretagogues. After mechanical and enzymatic dispersion, the human fetal pituitary cells were cultured on an extracellular matrixlike-material-coated 24-well plate. After 3 days' incubation in serum-containing medium, the PCCs were washed and preincubated for 90 min in serum-free medium and incubated with activin-A, inhibin-A, TGF-beta, follistatin, sex steroids, and GnRH, in quadruplicate. RESULTS: Activin-A was a potent secretagogue for FSH secretion. GnRH (20 ng/ml) was more potent than rh-activin-A for LH secretion. Nevertheless, a significant increase in LH secretion into the medium was brought about by rh-activin-A. Inhibin decreased FSH and LH secretion, but the LH response to inhibin was less prominent than that of FSH. GnRH opposed the inhibitory effect of inhibin on LH secretion. In dynamic, short-term, repetitive exposure of fetal pituitary fragments to rh-activin-A (superfusion), we could not receive a similar increase in LH and FSH as in static incubations, as opposed to a short GnRH exposure. In addition to their endocrine, paracrine, and autocrine effects, and in addition to their role as possible markers, the TGF-beta superfamily members may affect embryogenesis and possibly immunomodulation of the embryo and fetus. The role of activin and inhibin as intragonadal regulators is hypothesized. The pro-alphaC inhibin precursor may act as an FSH receptor antagonist. CONCLUSIONS: Human fetal PCCs express the previously reported physiologic responses to activin and inhibin generated in nonhuman experiments on gonadotropin secretion in vitro and may serve as a physiologic model for studying human gonadotrope responses to the TGF-beta family of peptides.
BACKGROUND:Activin has been previously demonstrated to directly stimulate the synthesis of gonadotropin-releasing hormone (GnRH) receptors and to increase follicle-stimulating hormone (FSH) secretion in nonhuman pituitary cell cultures (PCCs). Currently, knowledge of the physiological role of these peptides in primates is still far from complete. Moreover, several results in macaque monkeys failed to support an unequivocal role for inhibin in FSH suppression. Whereas the bioactivity of inhibin and activin has been demonstrated in rat PCCs, no data exist on human pituitary response to these peptides either in vivo or in vitro. METHODS: We studied the secretion of FSH and luteinizing hormone (LH) by dispersed human fetal pituitary cells from midtrimester abortions in response to recombinant human (rh-) activin-A, inhibin-A, and other secretagogues. After mechanical and enzymatic dispersion, the human fetal pituitary cells were cultured on an extracellular matrixlike-material-coated 24-well plate. After 3 days' incubation in serum-containing medium, the PCCs were washed and preincubated for 90 min in serum-free medium and incubated with activin-A, inhibin-A, TGF-beta, follistatin, sex steroids, and GnRH, in quadruplicate. RESULTS:Activin-A was a potent secretagogue for FSH secretion. GnRH (20 ng/ml) was more potent than rh-activin-A for LH secretion. Nevertheless, a significant increase in LH secretion into the medium was brought about by rh-activin-A. Inhibin decreased FSH and LH secretion, but the LH response to inhibin was less prominent than that of FSH. GnRH opposed the inhibitory effect of inhibin on LH secretion. In dynamic, short-term, repetitive exposure of fetal pituitary fragments to rh-activin-A (superfusion), we could not receive a similar increase in LH and FSH as in static incubations, as opposed to a short GnRH exposure. In addition to their endocrine, paracrine, and autocrine effects, and in addition to their role as possible markers, the TGF-beta superfamily members may affect embryogenesis and possibly immunomodulation of the embryo and fetus. The role of activin and inhibin as intragonadal regulators is hypothesized. The pro-alphaC inhibin precursor may act as an FSH receptor antagonist. CONCLUSIONS:Human fetal PCCs express the previously reported physiologic responses to activin and inhibin generated in nonhuman experiments on gonadotropin secretion in vitro and may serve as a physiologic model for studying human gonadotrope responses to the TGF-beta family of peptides.
Authors: Lacey L Roybal; Arpi Hambarchyan; Jason D Meadows; Nermeen H Barakat; Patricia A Pepa; Kellie M Breen; Pamela L Mellon; Djurdjica Coss Journal: Mol Endocrinol Date: 2014-08-08