Animal Reproduction Science 143 (2013) 30–37
Contents lists available at ScienceDirect
Animal Reproduction Science journal homepage: www.elsevier.com/locate/anireprosci
Involvement of the orphan nuclear receptor SF-1 in the effect of PCBs, DDT and DDE on the secretion of steroid hormones and oxytocin from bovine granulosa cells J. Mlynarczuk, M.H. Wrobel, A. Ziolkowska, J. Kotwica ∗ Institute of Animal Reproduction and Food Research of the Polish Academy of Sciences, Tuwima St 10, 10-747 Olsztyn, Poland
a r t i c l e
i n f o
Article history: Received 26 April 2013 Received in revised form 26 September 2013 Accepted 16 October 2013 Available online 26 October 2013 Keywords: SF-1 PCBs DDT Steroidogenesis Oxytocin Granulosa
a b s t r a c t Polychlorinated biphenyls (PCBs), DDT and its metabolite (DDE) belong to estrogen-like endocrine disruptors. However, though their activity is approximately 1000-fold lower than the activity of estradiol (E2), this steroid’s high concentration in follicular fluid and incubation media does not inhibit the influence of these xenobiotics. It was hypothesized that these xenobiotics might affect Steroidogenic Factor-1 (SF-1) and impair ovary function. To test this hypothesis, granulosa cells were obtained from ovarian follicles >1 or 0.05), except when FSH was used as a positive control (Fig. 2). All of the xenobiotics tested, as well as HxP, significantly (P < 0.05) increased the secretion of progesterone from granulosa cells that were derived from follicles 1 cm (Fig. 4A) and inhibited the stimulatory effect
of all xenobiotics and HxP in cell cultures derived from follicles 1 cm (P > 0.05, Fig. 5A). However, when given alone, SF-1 antagonist had no significant effect on the expression of SF-1 (P > 0.05). There were no significant (P > 0.05) changes in the mRNA expression of the SF-1 gene under the influence of the examined factors (Fig. 6). 4. Discussion The increase in P4 and OT secretion evoked by the xenobiotics used in this study confirms the data obtained in our
(A)
(B)
b bc
2
b bc
c
b c
2 d
ab a
ad
a
a
ad
a
1
b
b
cd d
a
a
a
d
1
D D T D D PC E B PC 77 B 15 3 H xP D D T D D PC E B PC 77 B 15 3
F0 16 0
xP H
D D T D D PC E B PC 77 B 15 3 H xP D D T D D PC E B PC 77 B 15 3
F0 16 0
trl
xP H
C
F0160-6M
trl
0
0
C
Arbitrary units
c
3
3
F0160-6M
Fig. 5. The relative expression (mean ± SEM, n = 4) of NP-I/OT mRNA in granulosa cells treated with xenobiotics (10 ng/ml) and with or without SF-1 inhibitor (F0160; 1 × 10−6 M). (A) Cells from follicles 1 cm. Control values accepted as 1. The bars with different superscripts are significantly different (P < 0.05).
J. Mlynarczuk et al. / Animal Reproduction Science 143 (2013) 30–37
(A)
(B)
b ab ab
ab
0.0
0.0 D D
D D
F0160 (1x10-6M)
ab
ab
ab
ab
a
C tr l H xP F0 16 0 D D T D D PC E B PC 77 B 15 3 D D T D D PC E B PC 77 B 15 3
0.5
PC E B PC 77 B 15 3
0.5
T
1.0
b ab
1.5
1.0
C tr l H xP F0 16 0 D D T D D PC E B PC 77 B 15 3
Arbitrary units
1.5
35
F0160 (1x10-6M)
Fig. 6. The relative expression (mean ± SEM, n = 4) of SF-1 mRNA in granulosa cells treated with xenobiotics (10 ng/ml) and with or without SF-1 inhibitor (F0160; 1 × 10−6 M). (A) Cells from follicles 1 cm. Control values accepted as 1. The bars with different superscripts are significantly different (P < 0.05).
earlier investigations (Mlynarczuk et al., 2005, 2009). However, the main aim of this work was to verify the hypothesis that SF-1 acts as an intermediate in mediating the effect xenobiotics on the synthesis and secretion of these hormones. Considering the low affinity of the xenobiotics used for the estrogen receptor (ER) (Matthews et al., 2000) on the one hand and the high concentrations of the native hormone in culture medium (200–500 pg/ml) on the other hand, the effect of xenoestrogens at the doses used should be blocked by the native hormone. The lack of observed effect of xenobiotics on the secretion of E2 may confirm this assumption. However, the measurement of E2 secretion alone may be an insufficient indicator of changes in a cell under the influence of xenobiotics, because it has been demonstrated in humans that the expression of the CYP19, which encodes aromatase, is under the partial control of SF-1 (Michael et al., 1995). Despite this, there were neither effect of xenobiotics nor HxP on E2 secretion. Presumably because the stimulation of CYP19 expression can also be evoked by another orphan receptor, i.e. LHR-1 (NR5A2) closely related to the SF-1 (Galarneau et al., 1996; Saxena et al., 2007). Furthermore, LHR-1 has a higher transcriptional activity than the SF-1 to stimulate the expression of CYP19 in the early stages of the follicular maturation. However, during follicle growth the roles of these receptors are reversed gradually, but only in the periovulatory phase the expression of CYP19 is under control of SF-1 (Hinshelwood et al., 2003). Hence, the impact of xenobiotics on the secretion of E2 could not be observed in the test model. In contrast, ovarian steroidogenic cells treated with xenobiotics secreted markedly higher amounts of P4 (Nejaty et al., 2001; Mlynarczuk et al., 2005). Presumably, this higher secretion occurs because SF-1 controls three key factors that are responsible for P4 synthesis: StAR protein, cytochrome P450scc and 3HSD (Clemens et al., 1994; Manna et al., 2002; Leers-Sucheta et al., 1997). This finding is clearly observed in the present study, particularly in cells that were treated with stimulator alone or stimulator given jointly with an SF-1 antagonist. However,
SF-1 antagonist does not completely inhibit the increased P4 secretion induced by xenobiotics, particularly PCB-77 congener. This observation can be explained by xenobiotics higher affinity for the SF-1 compared to the synthetic ligand HxP, which, although it is highly selective, has a relatively low affinity for this receptor (Del Tredici et al., 2008). It is therefore possible that the dose of antagonist observed to effectively inhibit the effect of HxP was too small to completely block the influence of xenobiotics on granulosa cells. Unfortunately, increasing the dose of antagonist over 1 × 10−6 M and giving it jointly with xenobiotics resulted in a significant decrease in the viability of tested cells. Conversely, the stimulation of SF-1 does not always result in an increase in the expression of steroidogenic pathway enzyme genes (Christenson and Devoto, 2003). To induce the expression of the genes for StAR protein or 3ˇHSD, SF-1 itself binds with its requisite coactivators, such as GATA4 or GATA6 (Jimenez et al., 2003; Viger et al., 2008). Moreover, other proteins, such as DAX-1 or WT-1, can also modulate the activity of this receptor (Nachtigal et al., 1998). Therefore, it appears that the activation of SF-1 at the same time as xenobiotic treatment impairs its interaction with co-activators in the steroidogenic cells. The blockade of SF-1 also inhibits the synthesis and secretion of OT, as stimulated by xenobiotic treatment. The increase of NP-I/OT gene expression in bovine ovary is dependent on SF-1 (Wehrenberg et al., 1994). This process requires the dimerization of SF-1 monomers, although it is not known whether co-activators are necessary to initiate this process. Thus, we observed a high efficiency of the inhibition of NP-I/OT gene expression, as induced by either HxP or xenobiotics, by F0160. This finding was especially apparent in cultured granulosa cells that were derived from ovarian follicles with a diameter of >1 cm. It should be noted that the amount of SF-1 protein rises during growth and maturation of ovarian follicles (Sadowsky and Crawford, 1998). As the ovarian follicles develop the synthesis of the OT precursor is rapidly intensified, however, OT
36
J. Mlynarczuk et al. / Animal Reproduction Science 143 (2013) 30–37
secretion varies slightly until LH surge (Voss and Fortune, 1992; Gimpl and Fahrenholz, 2001), and at the time it supports luteinization of granulosa cells (Tallam et al., 2000). Thus xenobiotics which impair OT synthesis, acting via SF1, may affect follicle growth and maturation. Although F0160 did not always inhibit OT secretion to control levels after stimulation, it should be remembered that xenobiotic treatment increases the expression of the peptidyl glycine-␣-amidating mono-oxygenase (PGA) gene in bovine luteal and granulosa cells (Mlynarczuk et al., 2009). This enzyme, whose expression is E2-dependent (Ivell and Walther, 1999) but independent of SF-1 (Downey and Donlon, 1997), is responsible for the post-translational processing of OT and the release of the active hormone from the cell. PCB-77 and DDE, which have estrogenlike properties, were the most efficient at increasing PGA gene expression in granulosa cells (Mlynarczuk et al., 2009). Therefore, though NP-I/OT gene expression and OT pro-hormone production were inhibited by F0160 after xenobiotic treatment (Fig. 5), it is likely that the increase in the amount of PGA evoked by xenobiotics could compensate for the expected decrease in the secretion of OT (Fig. 4). No changes were found in the expression of SF-1 mRNA in response to any of the xenobiotics used, suggesting that the observed effects of these xenobiotics is not due to the altered quantity of SF-1 in the cell, which would thereby change its sensitivity to the test substances. In conclusion, the data obtained indicate that the disrupting effects of xenobiotics on the synthesis and secretion of P4 and OT from granulosa cells and the consequent development and maturation of ovarian follicles, may take place via SF-1. Acknowledgments We thank Dr. G.L. Williams (Texas A&M University, Beeville, USA) for estradiol antiserum, Dr. G. Kotwica and Dr. S. Okrasa (University of Warmia and Mazury, Olsztyn, Poland) for oxytocin and progesterone antisera, respectively. FSH (AFP-4261-A) was purchased from Dr. A.F. Parlow (Pituitary Hormones & Antisera Center, Torrance, CA, USA). This study was supported by a grant (N/N308 006136) from the National Science Centre and by the Polish Academy of Sciences. References Barnhart, K.M., Mellon, P.L., 1994. The orphan nuclear receptor, SF1, regulates the glycoprotein hormone alpha-subunit in pituitary gonadotropes. Mol. Endocrinol. 8, 878–885. Benoit, G., Cooney, A., Giguere, V., Ingraham, H., Lazar, M., Muscat, G., Perlmann, T., Renaud, J.-P., Schwabe, J., Sladek, F., Tsai, M.-J., Laudet, V., 2006. International union of pharmacology LXVI. Orphan nuclear receptors. Pharmacol. Rev. 58, 798–836. Brendtson, A.K., Weaver, C.J., Fortune, J.E., 1996. Differential effects of oxytocin on steroid production by bovine granulosa cells. Mol. Cell. Endocrinol. 116, 191–198. Chomczynski, P., Sacchi, N., 1987. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-cloroform extraction. Anal. Biochem. 162, 156–159. Christenson, L.K., Devoto, L., 2003. Cholesterol transport and steroidogenesis by the corpus luteum. Reprod. Biol. Endocrinol. 1, 90. Clemens, J.W., Lala, D.S., Parker, K.L., Richards, J.S., 1994. Steroidogenic Factor-1 binding and transcriptional activity of the cholesterol
side-chain cleavage promoter in rat granulosa cells. Endocrinology 134, 1499–1508. Cogliano, V.J., 1998. Assessing the cancer risk from environmental PCBs. Environ. Health Perspect. 101, 378–384. Corona-Cruz, A., Gold-Buchot, G., Gutierrez-Rojas, M., MonroyHermosillo, O., Favela, E., 1999. Anaerobic–aerobic biodegradation of DDT (dichlorodiphenyl trichloroethane) in soil. Bull. Environ. Contam. Toxicol. 63, 219–225. Del Tredici, A.L., Andersen, C.B., Currier, E.A., Ohrmund, S.R., Fairbain, L.C., Lund, B.W., Nash, N., Olsson, R., Piu, F., 2008. Identification of the first synthetic steroidogenic factor 1 inverse agonists: pharmacological modulation of steroidogenic enzymes. Mol. Pharmacol. 73, 900–908. Downey, E., Donlon, J., 1997. Identity of bovine growth hormone and peptidylglycine monooxygenase. Arch. Biochem. Biophys. 345, 193–198. Durlinger, A.L., Kramer, P., Karels, B., de Jong, F.H., Uilenbroek, J.T., Grootegoed, J.A., Themmen, A.P., 1999. Control of primordial follicle recruitment by anti-Müllerian hormone in the mouse ovary. Endocrinology 140, 5789–5796. Falender, A.E., Lanz, R., Malefant, D., Belanger, L., Richards, J.S., 2003. Differential expression of steroidogenic factor-1 and FTF/LRH-1 in the rodent ovary. Endocrinology 144, 3598–3610. Fan, Q.W., Yanase, T., Morinaga, H., Gondo, S., Okabe, T., Nomura, M., Komatsu, T., Morohashi, K.-I., Hayes, T.B., Takayanagi, R., Nawata, H., 2007. Atrazine-induced aromatase expression is SF-1 dependent: implications for endocrine disruptions in wildlife and reproductive cancers in human. Environ. Health Perspect. 115, 720–727. Galarneau, L., Paré, J.F., Allard, D., Hamel, D., Levesque, L., Tugwood, J.D., Green, S., Bélanger, L., 1996. The alpha1-fetoprotein locus is activated by a nuclear receptor of the Drosophila FTZ-F1 family. Mol. Cell. Biol. 16, 3853–3865. Gimpl, G., Fahrenholz, F., 2001. The oxytocin receptor system: structure, function, and regulation. Physiol. Rev. 8, 629–683. Hinshelwood, M.M., Repa, J.J., Shelton, J.M., Richardson, J.A., Mangelsdorf, D.J., Mendelson, C.R., 2003. Expression of LRH-1 and SF-1 in the mouse ovary: localization in different cell types correlates with differing function. Mol. Cell. Endocrinol. 207, 39–45. Ho, P.W., Garner, C.E., Ho, J.W., Leung, K.C., Chu, A.C., Kwok, K.H., Kung, M.H., Burka, L.T., Ramsden, D.B., Ho, S.L., 2008. Estrogenic phenol and catechol metabolites of PCBs modulate catechol-O-methyltransferase expression via the estrogen receptor: potential contribution to cancer risk. Curr. Drug Metab. 9, 304–309. Hodgert Jury, H., Zacharewski, T.R., Hammond, G.L., 2000. Interactions between human plasma sex hormone-binding globulin and xenobiotic ligands. J. Steroid Biochem. 75, 167–175. Holvorson, L.H., Kaiser, U.B., Chin, W.W., 1996. Stimulation of luteinizing hormone  gene promoter activity by the orphan receptor, steroidogenic factor-1. J. Biol. Chem. 271, 6645–6650. Honda, T., Morohashi, K.I., Nomura, T., Takeya, K., Kitajima, M., Omura, T., 1993. Ad4BP regulating steroidogenic P450 gene is a member of steroid hormone receptor superfamily. J. Biol. Chem. 268, 7494–7502. Ivell, R., Walther, N., 1999. The role of sex steroids in the oxytocin hormone system. Mol. Cell. Endocrinol. 155, 95–101. Jeyasuria, P., Ikeda, Y., Jasmin, S.P., Zhao, L., Deb Rooij, D.G., Themmen, A.P., Behringer, R.R., Parker, K.L., 2004. Cell-specific knockout steroidogenic factor 1 reveals is essential role in gonadal function. Mol. Endocrinol. 18, 1610–1619. Jimenez, P., Saner, K., Mayhew, B., Rainey, W.E., 2003. GATA-6 is expressed in the human adrenal and regulates transcription of genes required for adrenal biosynthesis. Endocrinology 144, 4285–4288. Jo, M., Fortune, J.E., 2002. Oxytocin inhibits LH-stimulated production of androstendione by bovine theca cells. Mol. Cell. Endocrinol. 188, 151–159. Kotwica, J., Skarzynski, D., 1993. Influence of oxytocin removal from corpus luteum on secretory function and duration of the oestrous cycle in cattle. J. Reprod. Fertil. 97, 411–417. Kotwica, J., Skarzynski, D.J., Jaroszewski, J.J., Bogacki, M., 1994. Noradrenaline affects secretory function of corpus luteum independently on prostaglandins in conscious cattle. Prostaglandins 48, 1–10. Landau, S., Braw-Tal, R., Kaim, L., Bor, A., Bruckental, I., 2000. Preovulatory follicular status and diet affect the insulin and glucose content of follicles in high-yielding dairy cows. Anim. Reprod. Sci. 64, 181–197. Leers-Sucheta, S., Morohashi, K., Mason, J., Melner, M., 1997. Synergistic activation of the human type II 3HSD/D5-D4 isomerase promoter by the transcriptional factor SF-1/Ad4BP and phorbol ester. J. Biol. Chem. 272, 7960–7967. Li, J., Li, N., Ma, M., Giesy, J.P., Wang, Z., 2008. In vitro profiling of the endocrine disrupting potency of organochlorine pesticides. Toxicol. Lett. 183, 65–71.
J. Mlynarczuk et al. / Animal Reproduction Science 143 (2013) 30–37 Madoux, F., Li, X., Chase, P., Zastrow, G., Cameron, M.D., Conkright, J.J., Griffin, P.R., Thacher, S., Hodder, P., 2008. Potent, selective and cell penetrant inhibitors of SF-1 by functional uHTS. Mol. Pharmacol. 73, 1776–1784. Manna, P.R., Dyson, M.T., Eubank, D.W., Clark, B.J., Lalli, E., Sassone-Corsi, P., Zeleznik, A.J., Stocco, D.M., 2002. Regulatory of steroidogenesis and the steroidogenic acute protein by a member of the cAMP responseelement binding protein family. Mol. Endocrinol. 16, 184–199. Marchal, R., Naville, D., Durand, P., Bergot, M., Phenoat, A., 1998. A steroidogenic factor-1 binding element is essential for basal human ACTH receptor gene transcription. Biochem. Biophys. Res. Commun. 247, 28–32. Matthews, J., Celius, T., Halgren, L., Zacharewski, T., 2000. Differential estrogen receptor binding of estrogenic substances: a species comparison. J. Steroid Biochem. 74, 223–234. Michael, M.D., Kligore, M.W., Morohashi, K.I., Simpson, E.R., 1995. Ad4BP/SF-1 regulates cAMP-induced transcription from the proximal promoter (PII) of the human aromatase P450 (CYP19) gene in ovary. J. Biol. Chem. 270, 13561–13566. Młynarczuk, J., Kotwica, J., 2005. Influence of polichlorinated biphenyles on the secretion of oxytocin from bovine luteal cells and from granulosa cells obtained from the follicles of different size. Pol. J. Vet. Sci. 8, 261–267. Mlynarczuk, J., Rekawiecki, R., 2010. The role of thr orphan receptor SF-1 in the development and function of the ovary. Reprod. Biol. 10, 177–193. Młynarczuk, J., Niewiadowska, A., Kotwica, J., 2005. Polychlorinated biphenyls impaired FSH-stimulated on the secretion of steroids and oxytocin from bovine granulosa cells. Bull. Vet. Inst. Puławy 49, 411–417. Mlynarczuk, J., Wrobel, M., Kotwica, J., 2009. The influence of polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane (DDT) and its metabolite – dichlorodiphenyldichloroethylene (DDE) on mRNA expression for NP-I/OT and PGA, involved in oxytocin synthesis in bovine granulosa and luteal cells. Reprod. Toxicol. 28, 354–358. Mlynarczuk, J., Wrobel, M.H., Rekawiecki, R., Kotwica, J., 2013. The expression of Steroidogenic Factor-1 and its role in bovine steroidogenic ovarian cells during the estrus cycle and first trimester of pregnancy. Anim. Reprod. Sci. 138, 74–81. Morohashi, K.I., Honda, S.-I., Inomata, Y., Handa, H., Omura, T., 1992. A common trans-acting factor, Ad4-binding protein, to the promoters of steroidogenic P450-s. J. Biol. Chem. 267, 17913–17919. Nachtigal, M.W., Hirokawa, Y., Enyeart-VanHouten, D.L., Flanagan, J.N., Hammer, G.D., Ingraham, H.A., 1998. Wilms’ tumor 1 and Dax-1 modulate the orphan nuclear receptor SF-1 in sex specific gene expression. Cell 93, 445–454. Nejaty, H., Lacey, M., Whitehead, S.A., 2001. Differing effects of endocrinedisrupting chemicals on basal and FSH-stimulated progesterone production in rat granulosa-luteal cells. Exp. Biol. Med. 226, 570–576. Nesaretnam, K., Corcoran, D., Dills, R.R., Darbre, P., 1996. 3,3 ,4,4 Tetrachlorobiphenyl acts an estrogen in vitro and in vivo. Mol. Endocrinol. 10, 923–936. Okuda, K., Uenoyama, Y., Fujita, Y., Iga, K., Sakamoto, K., Kimura, T., 1997. Functional oxytocin receptors in bovine granulosa cells. Biol. Reprod. 56, 625–631. Ozisik, G., Achermann, J.C., Jameson, J.L., 2002. The role of SF-1 in adrenal and reproductive functions: insight from naturally occurring mutations in humans. Mol. Genet. Metab. 76, 85–91. Parker, K.L., Rice, D.A., Lala, D.S., Ikeda, Y., Luo, X., Bakke, M., Zhao, L., Frigieri, C., Hanley, N.A., Stallings, N., Schimmer, B.P., 2002. Steroidogenic factor 1: an essential mediator of endocrine development. Recent Prog. Horm. Res. 57, 19–39.
37
Pogrmic-Majkic, K., Fa, S., Dakic, V., Kaisarevic, S., Kovacevic, R., 2010. Upregulation of peripubertal rat Leydig cell steroidogenesis following 24 h in vitro and in vivo exposure to atrazine. Toxicol. Sci. 118, 52–60. Prakash, B.S., Meyer, H.D., Schallenberger, E., Van de Wiel, D.F., 1987. Development of sensitive enzyme immunoassay (EIA) for progesterone determination in unextracted bovine plasma using the second antibody technique. J. Steroid Biochem. 28, 623–627. Roselli, M., Reinhart, K., Imthurn, B., Keller, P.J., Dubey, R.K., 2000. Cellular and biochemical mechanisms by which environmental estrogens influence reproductive function. Hum. Reprod. Update 6, 332–350. Sadowsky, Y., Crawford, P.A., 1998. Developmental and physiologic roles of the nuclear receptor Steroidogenic Factor-I in the reproductive system. J. Soc. Gynecol. Investig. 5, 6–12. Safe, S.H., Pallaroni, L., Yoon, K., Gaido, K., Ross, S., Saville, B., McDonnell, D., 2001. Toxicology of environmental estrogens. Reprod. Fertil. Dev. 13, 307–315. Saxena, D., Escamilla-Hernandez, R., Little-Ihring, L., Zeleznik, A.J., 2007. Liver receptor homolog-1 and steroidogenic factor-1 have similar actions on rat granulosa cell steroidogenesis. Endocrinology 148, 726–734. Shen, W.H., Moore, C.C.D., Ikeda, Y., Parker, K.L., Ingraham, H.A., 1994. Nuclear receptor steroidogenic factor 1 regulates the Müllerian inhibiting substance gene: a link to the sex determination cascade. Cell 77, 651–661. Tallam, S.K., Walton, J.S., Johnson, W.H., 2000. Effects of oxytocin on follicular development and duration of the estrous cycle in heifers. Theriogenology 53, 951–962. Viger, R.S., Guittot, S.M., Anttonen, M., Wilson, D.B., Heikinheimo, M., 2008. Role of the GATA family of transcription factors in endocrine development, function, and disease. Mol. Endocrinol. 22, 781–798. Voss, A.K., Fortune, J.E., 1992. Oxytocin/neurophysin-I messenger ribonucleic acid in bovine granulosa cells increases after the luteinizing hormone (LH) surge and is stimulated by LH in vitro. Endocrinology 131, 2755–2762. Walther, N., Jansen, M., Akbary, W., Ivell, R., 2006. Differentiation-specific action of orphan nuclear receptor NR5A1 (SF-1): transcriptional regulation in luteinizing bovine theca cells. Reprod. Biol. Endocrinol. 4, 64. Wehrenberg, U., Ivell, R., Jansen, M., von Goedecke, S., Walther, N., 1994. Two orphan receptors binding to a common site are involved in the regulation of the oxytocin gene in the bovine ovary. Proc. Natl. Acad. Sci. U. S. A. 91, 1440–1444. Wilson., V.S., Blystone, C.R., Hotchkiss, A.K., Rider, C.V., Gray Jr., L.E., 2008. Diverse mechanisms of anti-androgen action: impact on male rat reproductive tract development. Int. J. Androl. 31, 178–187. Wojtowicz, A., Tomanek, M., Augustowska, K., Gregoraszczuk, E.L., 2005. Aromatic hydrocarbon receptor (AhR) in the porcine theca and granulosa cells: effect of TCDD, PCB 126 and PCB 153 on the expression of AhR. Endocrine Reg. 39, 109–118. Wrobel, M., Mlynarczuk, J., Kotwica, J., 2009. The adverse effect of dihydrodiphenyltrichloroetane (DDT) and its metabolite (DDE) on the secrertion of prostaglandins and oxytocin in bovine cultured ovarian and endometrial cells. Reprod. Toxicol. 27, 72–78. Zhang, P., Mellon, S.H., 1996. The orphan nuclear receptor steroidogenic factor-1 regulates the cAMP mediated transcriptional activation of rat cytochrome P450c14 (17␣-hydroxylase/c17-20 lyase). Mol. Endocrinol. 10, 147–158. Zhao, L., Bakke, M., Krimkevich, Y., Cushman, L.J., Parlow, A.F., Camper, S.A., Parker, K.L., 2001. Steroidogenic factor 1 (SF1) is essential for pituitary gonadotrope function. Development 128, 147–154.
Contents lists available at ScienceDirect
Animal Reproduction Science journal homepage: www.elsevier.com/locate/anireprosci
Involvement of the orphan nuclear receptor SF-1 in the effect of PCBs, DDT and DDE on the secretion of steroid hormones and oxytocin from bovine granulosa cells J. Mlynarczuk, M.H. Wrobel, A. Ziolkowska, J. Kotwica ∗ Institute of Animal Reproduction and Food Research of the Polish Academy of Sciences, Tuwima St 10, 10-747 Olsztyn, Poland
a r t i c l e
i n f o
Article history: Received 26 April 2013 Received in revised form 26 September 2013 Accepted 16 October 2013 Available online 26 October 2013 Keywords: SF-1 PCBs DDT Steroidogenesis Oxytocin Granulosa
a b s t r a c t Polychlorinated biphenyls (PCBs), DDT and its metabolite (DDE) belong to estrogen-like endocrine disruptors. However, though their activity is approximately 1000-fold lower than the activity of estradiol (E2), this steroid’s high concentration in follicular fluid and incubation media does not inhibit the influence of these xenobiotics. It was hypothesized that these xenobiotics might affect Steroidogenic Factor-1 (SF-1) and impair ovary function. To test this hypothesis, granulosa cells were obtained from ovarian follicles >1 or 0.05), except when FSH was used as a positive control (Fig. 2). All of the xenobiotics tested, as well as HxP, significantly (P < 0.05) increased the secretion of progesterone from granulosa cells that were derived from follicles 1 cm (Fig. 4A) and inhibited the stimulatory effect
of all xenobiotics and HxP in cell cultures derived from follicles 1 cm (P > 0.05, Fig. 5A). However, when given alone, SF-1 antagonist had no significant effect on the expression of SF-1 (P > 0.05). There were no significant (P > 0.05) changes in the mRNA expression of the SF-1 gene under the influence of the examined factors (Fig. 6). 4. Discussion The increase in P4 and OT secretion evoked by the xenobiotics used in this study confirms the data obtained in our
(A)
(B)
b bc
2
b bc
c
b c
2 d
ab a
ad
a
a
ad
a
1
b
b
cd d
a
a
a
d
1
D D T D D PC E B PC 77 B 15 3 H xP D D T D D PC E B PC 77 B 15 3
F0 16 0
xP H
D D T D D PC E B PC 77 B 15 3 H xP D D T D D PC E B PC 77 B 15 3
F0 16 0
trl
xP H
C
F0160-6M
trl
0
0
C
Arbitrary units
c
3
3
F0160-6M
Fig. 5. The relative expression (mean ± SEM, n = 4) of NP-I/OT mRNA in granulosa cells treated with xenobiotics (10 ng/ml) and with or without SF-1 inhibitor (F0160; 1 × 10−6 M). (A) Cells from follicles 1 cm. Control values accepted as 1. The bars with different superscripts are significantly different (P < 0.05).
J. Mlynarczuk et al. / Animal Reproduction Science 143 (2013) 30–37
(A)
(B)
b ab ab
ab
0.0
0.0 D D
D D
F0160 (1x10-6M)
ab
ab
ab
ab
a
C tr l H xP F0 16 0 D D T D D PC E B PC 77 B 15 3 D D T D D PC E B PC 77 B 15 3
0.5
PC E B PC 77 B 15 3
0.5
T
1.0
b ab
1.5
1.0
C tr l H xP F0 16 0 D D T D D PC E B PC 77 B 15 3
Arbitrary units
1.5
35
F0160 (1x10-6M)
Fig. 6. The relative expression (mean ± SEM, n = 4) of SF-1 mRNA in granulosa cells treated with xenobiotics (10 ng/ml) and with or without SF-1 inhibitor (F0160; 1 × 10−6 M). (A) Cells from follicles 1 cm. Control values accepted as 1. The bars with different superscripts are significantly different (P < 0.05).
earlier investigations (Mlynarczuk et al., 2005, 2009). However, the main aim of this work was to verify the hypothesis that SF-1 acts as an intermediate in mediating the effect xenobiotics on the synthesis and secretion of these hormones. Considering the low affinity of the xenobiotics used for the estrogen receptor (ER) (Matthews et al., 2000) on the one hand and the high concentrations of the native hormone in culture medium (200–500 pg/ml) on the other hand, the effect of xenoestrogens at the doses used should be blocked by the native hormone. The lack of observed effect of xenobiotics on the secretion of E2 may confirm this assumption. However, the measurement of E2 secretion alone may be an insufficient indicator of changes in a cell under the influence of xenobiotics, because it has been demonstrated in humans that the expression of the CYP19, which encodes aromatase, is under the partial control of SF-1 (Michael et al., 1995). Despite this, there were neither effect of xenobiotics nor HxP on E2 secretion. Presumably because the stimulation of CYP19 expression can also be evoked by another orphan receptor, i.e. LHR-1 (NR5A2) closely related to the SF-1 (Galarneau et al., 1996; Saxena et al., 2007). Furthermore, LHR-1 has a higher transcriptional activity than the SF-1 to stimulate the expression of CYP19 in the early stages of the follicular maturation. However, during follicle growth the roles of these receptors are reversed gradually, but only in the periovulatory phase the expression of CYP19 is under control of SF-1 (Hinshelwood et al., 2003). Hence, the impact of xenobiotics on the secretion of E2 could not be observed in the test model. In contrast, ovarian steroidogenic cells treated with xenobiotics secreted markedly higher amounts of P4 (Nejaty et al., 2001; Mlynarczuk et al., 2005). Presumably, this higher secretion occurs because SF-1 controls three key factors that are responsible for P4 synthesis: StAR protein, cytochrome P450scc and 3HSD (Clemens et al., 1994; Manna et al., 2002; Leers-Sucheta et al., 1997). This finding is clearly observed in the present study, particularly in cells that were treated with stimulator alone or stimulator given jointly with an SF-1 antagonist. However,
SF-1 antagonist does not completely inhibit the increased P4 secretion induced by xenobiotics, particularly PCB-77 congener. This observation can be explained by xenobiotics higher affinity for the SF-1 compared to the synthetic ligand HxP, which, although it is highly selective, has a relatively low affinity for this receptor (Del Tredici et al., 2008). It is therefore possible that the dose of antagonist observed to effectively inhibit the effect of HxP was too small to completely block the influence of xenobiotics on granulosa cells. Unfortunately, increasing the dose of antagonist over 1 × 10−6 M and giving it jointly with xenobiotics resulted in a significant decrease in the viability of tested cells. Conversely, the stimulation of SF-1 does not always result in an increase in the expression of steroidogenic pathway enzyme genes (Christenson and Devoto, 2003). To induce the expression of the genes for StAR protein or 3ˇHSD, SF-1 itself binds with its requisite coactivators, such as GATA4 or GATA6 (Jimenez et al., 2003; Viger et al., 2008). Moreover, other proteins, such as DAX-1 or WT-1, can also modulate the activity of this receptor (Nachtigal et al., 1998). Therefore, it appears that the activation of SF-1 at the same time as xenobiotic treatment impairs its interaction with co-activators in the steroidogenic cells. The blockade of SF-1 also inhibits the synthesis and secretion of OT, as stimulated by xenobiotic treatment. The increase of NP-I/OT gene expression in bovine ovary is dependent on SF-1 (Wehrenberg et al., 1994). This process requires the dimerization of SF-1 monomers, although it is not known whether co-activators are necessary to initiate this process. Thus, we observed a high efficiency of the inhibition of NP-I/OT gene expression, as induced by either HxP or xenobiotics, by F0160. This finding was especially apparent in cultured granulosa cells that were derived from ovarian follicles with a diameter of >1 cm. It should be noted that the amount of SF-1 protein rises during growth and maturation of ovarian follicles (Sadowsky and Crawford, 1998). As the ovarian follicles develop the synthesis of the OT precursor is rapidly intensified, however, OT
36
J. Mlynarczuk et al. / Animal Reproduction Science 143 (2013) 30–37
secretion varies slightly until LH surge (Voss and Fortune, 1992; Gimpl and Fahrenholz, 2001), and at the time it supports luteinization of granulosa cells (Tallam et al., 2000). Thus xenobiotics which impair OT synthesis, acting via SF1, may affect follicle growth and maturation. Although F0160 did not always inhibit OT secretion to control levels after stimulation, it should be remembered that xenobiotic treatment increases the expression of the peptidyl glycine-␣-amidating mono-oxygenase (PGA) gene in bovine luteal and granulosa cells (Mlynarczuk et al., 2009). This enzyme, whose expression is E2-dependent (Ivell and Walther, 1999) but independent of SF-1 (Downey and Donlon, 1997), is responsible for the post-translational processing of OT and the release of the active hormone from the cell. PCB-77 and DDE, which have estrogenlike properties, were the most efficient at increasing PGA gene expression in granulosa cells (Mlynarczuk et al., 2009). Therefore, though NP-I/OT gene expression and OT pro-hormone production were inhibited by F0160 after xenobiotic treatment (Fig. 5), it is likely that the increase in the amount of PGA evoked by xenobiotics could compensate for the expected decrease in the secretion of OT (Fig. 4). No changes were found in the expression of SF-1 mRNA in response to any of the xenobiotics used, suggesting that the observed effects of these xenobiotics is not due to the altered quantity of SF-1 in the cell, which would thereby change its sensitivity to the test substances. In conclusion, the data obtained indicate that the disrupting effects of xenobiotics on the synthesis and secretion of P4 and OT from granulosa cells and the consequent development and maturation of ovarian follicles, may take place via SF-1. Acknowledgments We thank Dr. G.L. Williams (Texas A&M University, Beeville, USA) for estradiol antiserum, Dr. G. Kotwica and Dr. S. Okrasa (University of Warmia and Mazury, Olsztyn, Poland) for oxytocin and progesterone antisera, respectively. FSH (AFP-4261-A) was purchased from Dr. A.F. Parlow (Pituitary Hormones & Antisera Center, Torrance, CA, USA). This study was supported by a grant (N/N308 006136) from the National Science Centre and by the Polish Academy of Sciences. References Barnhart, K.M., Mellon, P.L., 1994. The orphan nuclear receptor, SF1, regulates the glycoprotein hormone alpha-subunit in pituitary gonadotropes. Mol. Endocrinol. 8, 878–885. Benoit, G., Cooney, A., Giguere, V., Ingraham, H., Lazar, M., Muscat, G., Perlmann, T., Renaud, J.-P., Schwabe, J., Sladek, F., Tsai, M.-J., Laudet, V., 2006. International union of pharmacology LXVI. Orphan nuclear receptors. Pharmacol. Rev. 58, 798–836. Brendtson, A.K., Weaver, C.J., Fortune, J.E., 1996. Differential effects of oxytocin on steroid production by bovine granulosa cells. Mol. Cell. Endocrinol. 116, 191–198. Chomczynski, P., Sacchi, N., 1987. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-cloroform extraction. Anal. Biochem. 162, 156–159. Christenson, L.K., Devoto, L., 2003. Cholesterol transport and steroidogenesis by the corpus luteum. Reprod. Biol. Endocrinol. 1, 90. Clemens, J.W., Lala, D.S., Parker, K.L., Richards, J.S., 1994. Steroidogenic Factor-1 binding and transcriptional activity of the cholesterol
side-chain cleavage promoter in rat granulosa cells. Endocrinology 134, 1499–1508. Cogliano, V.J., 1998. Assessing the cancer risk from environmental PCBs. Environ. Health Perspect. 101, 378–384. Corona-Cruz, A., Gold-Buchot, G., Gutierrez-Rojas, M., MonroyHermosillo, O., Favela, E., 1999. Anaerobic–aerobic biodegradation of DDT (dichlorodiphenyl trichloroethane) in soil. Bull. Environ. Contam. Toxicol. 63, 219–225. Del Tredici, A.L., Andersen, C.B., Currier, E.A., Ohrmund, S.R., Fairbain, L.C., Lund, B.W., Nash, N., Olsson, R., Piu, F., 2008. Identification of the first synthetic steroidogenic factor 1 inverse agonists: pharmacological modulation of steroidogenic enzymes. Mol. Pharmacol. 73, 900–908. Downey, E., Donlon, J., 1997. Identity of bovine growth hormone and peptidylglycine monooxygenase. Arch. Biochem. Biophys. 345, 193–198. Durlinger, A.L., Kramer, P., Karels, B., de Jong, F.H., Uilenbroek, J.T., Grootegoed, J.A., Themmen, A.P., 1999. Control of primordial follicle recruitment by anti-Müllerian hormone in the mouse ovary. Endocrinology 140, 5789–5796. Falender, A.E., Lanz, R., Malefant, D., Belanger, L., Richards, J.S., 2003. Differential expression of steroidogenic factor-1 and FTF/LRH-1 in the rodent ovary. Endocrinology 144, 3598–3610. Fan, Q.W., Yanase, T., Morinaga, H., Gondo, S., Okabe, T., Nomura, M., Komatsu, T., Morohashi, K.-I., Hayes, T.B., Takayanagi, R., Nawata, H., 2007. Atrazine-induced aromatase expression is SF-1 dependent: implications for endocrine disruptions in wildlife and reproductive cancers in human. Environ. Health Perspect. 115, 720–727. Galarneau, L., Paré, J.F., Allard, D., Hamel, D., Levesque, L., Tugwood, J.D., Green, S., Bélanger, L., 1996. The alpha1-fetoprotein locus is activated by a nuclear receptor of the Drosophila FTZ-F1 family. Mol. Cell. Biol. 16, 3853–3865. Gimpl, G., Fahrenholz, F., 2001. The oxytocin receptor system: structure, function, and regulation. Physiol. Rev. 8, 629–683. Hinshelwood, M.M., Repa, J.J., Shelton, J.M., Richardson, J.A., Mangelsdorf, D.J., Mendelson, C.R., 2003. Expression of LRH-1 and SF-1 in the mouse ovary: localization in different cell types correlates with differing function. Mol. Cell. Endocrinol. 207, 39–45. Ho, P.W., Garner, C.E., Ho, J.W., Leung, K.C., Chu, A.C., Kwok, K.H., Kung, M.H., Burka, L.T., Ramsden, D.B., Ho, S.L., 2008. Estrogenic phenol and catechol metabolites of PCBs modulate catechol-O-methyltransferase expression via the estrogen receptor: potential contribution to cancer risk. Curr. Drug Metab. 9, 304–309. Hodgert Jury, H., Zacharewski, T.R., Hammond, G.L., 2000. Interactions between human plasma sex hormone-binding globulin and xenobiotic ligands. J. Steroid Biochem. 75, 167–175. Holvorson, L.H., Kaiser, U.B., Chin, W.W., 1996. Stimulation of luteinizing hormone  gene promoter activity by the orphan receptor, steroidogenic factor-1. J. Biol. Chem. 271, 6645–6650. Honda, T., Morohashi, K.I., Nomura, T., Takeya, K., Kitajima, M., Omura, T., 1993. Ad4BP regulating steroidogenic P450 gene is a member of steroid hormone receptor superfamily. J. Biol. Chem. 268, 7494–7502. Ivell, R., Walther, N., 1999. The role of sex steroids in the oxytocin hormone system. Mol. Cell. Endocrinol. 155, 95–101. Jeyasuria, P., Ikeda, Y., Jasmin, S.P., Zhao, L., Deb Rooij, D.G., Themmen, A.P., Behringer, R.R., Parker, K.L., 2004. Cell-specific knockout steroidogenic factor 1 reveals is essential role in gonadal function. Mol. Endocrinol. 18, 1610–1619. Jimenez, P., Saner, K., Mayhew, B., Rainey, W.E., 2003. GATA-6 is expressed in the human adrenal and regulates transcription of genes required for adrenal biosynthesis. Endocrinology 144, 4285–4288. Jo, M., Fortune, J.E., 2002. Oxytocin inhibits LH-stimulated production of androstendione by bovine theca cells. Mol. Cell. Endocrinol. 188, 151–159. Kotwica, J., Skarzynski, D., 1993. Influence of oxytocin removal from corpus luteum on secretory function and duration of the oestrous cycle in cattle. J. Reprod. Fertil. 97, 411–417. Kotwica, J., Skarzynski, D.J., Jaroszewski, J.J., Bogacki, M., 1994. Noradrenaline affects secretory function of corpus luteum independently on prostaglandins in conscious cattle. Prostaglandins 48, 1–10. Landau, S., Braw-Tal, R., Kaim, L., Bor, A., Bruckental, I., 2000. Preovulatory follicular status and diet affect the insulin and glucose content of follicles in high-yielding dairy cows. Anim. Reprod. Sci. 64, 181–197. Leers-Sucheta, S., Morohashi, K., Mason, J., Melner, M., 1997. Synergistic activation of the human type II 3HSD/D5-D4 isomerase promoter by the transcriptional factor SF-1/Ad4BP and phorbol ester. J. Biol. Chem. 272, 7960–7967. Li, J., Li, N., Ma, M., Giesy, J.P., Wang, Z., 2008. In vitro profiling of the endocrine disrupting potency of organochlorine pesticides. Toxicol. Lett. 183, 65–71.
J. Mlynarczuk et al. / Animal Reproduction Science 143 (2013) 30–37 Madoux, F., Li, X., Chase, P., Zastrow, G., Cameron, M.D., Conkright, J.J., Griffin, P.R., Thacher, S., Hodder, P., 2008. Potent, selective and cell penetrant inhibitors of SF-1 by functional uHTS. Mol. Pharmacol. 73, 1776–1784. Manna, P.R., Dyson, M.T., Eubank, D.W., Clark, B.J., Lalli, E., Sassone-Corsi, P., Zeleznik, A.J., Stocco, D.M., 2002. Regulatory of steroidogenesis and the steroidogenic acute protein by a member of the cAMP responseelement binding protein family. Mol. Endocrinol. 16, 184–199. Marchal, R., Naville, D., Durand, P., Bergot, M., Phenoat, A., 1998. A steroidogenic factor-1 binding element is essential for basal human ACTH receptor gene transcription. Biochem. Biophys. Res. Commun. 247, 28–32. Matthews, J., Celius, T., Halgren, L., Zacharewski, T., 2000. Differential estrogen receptor binding of estrogenic substances: a species comparison. J. Steroid Biochem. 74, 223–234. Michael, M.D., Kligore, M.W., Morohashi, K.I., Simpson, E.R., 1995. Ad4BP/SF-1 regulates cAMP-induced transcription from the proximal promoter (PII) of the human aromatase P450 (CYP19) gene in ovary. J. Biol. Chem. 270, 13561–13566. Młynarczuk, J., Kotwica, J., 2005. Influence of polichlorinated biphenyles on the secretion of oxytocin from bovine luteal cells and from granulosa cells obtained from the follicles of different size. Pol. J. Vet. Sci. 8, 261–267. Mlynarczuk, J., Rekawiecki, R., 2010. The role of thr orphan receptor SF-1 in the development and function of the ovary. Reprod. Biol. 10, 177–193. Młynarczuk, J., Niewiadowska, A., Kotwica, J., 2005. Polychlorinated biphenyls impaired FSH-stimulated on the secretion of steroids and oxytocin from bovine granulosa cells. Bull. Vet. Inst. Puławy 49, 411–417. Mlynarczuk, J., Wrobel, M., Kotwica, J., 2009. The influence of polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane (DDT) and its metabolite – dichlorodiphenyldichloroethylene (DDE) on mRNA expression for NP-I/OT and PGA, involved in oxytocin synthesis in bovine granulosa and luteal cells. Reprod. Toxicol. 28, 354–358. Mlynarczuk, J., Wrobel, M.H., Rekawiecki, R., Kotwica, J., 2013. The expression of Steroidogenic Factor-1 and its role in bovine steroidogenic ovarian cells during the estrus cycle and first trimester of pregnancy. Anim. Reprod. Sci. 138, 74–81. Morohashi, K.I., Honda, S.-I., Inomata, Y., Handa, H., Omura, T., 1992. A common trans-acting factor, Ad4-binding protein, to the promoters of steroidogenic P450-s. J. Biol. Chem. 267, 17913–17919. Nachtigal, M.W., Hirokawa, Y., Enyeart-VanHouten, D.L., Flanagan, J.N., Hammer, G.D., Ingraham, H.A., 1998. Wilms’ tumor 1 and Dax-1 modulate the orphan nuclear receptor SF-1 in sex specific gene expression. Cell 93, 445–454. Nejaty, H., Lacey, M., Whitehead, S.A., 2001. Differing effects of endocrinedisrupting chemicals on basal and FSH-stimulated progesterone production in rat granulosa-luteal cells. Exp. Biol. Med. 226, 570–576. Nesaretnam, K., Corcoran, D., Dills, R.R., Darbre, P., 1996. 3,3 ,4,4 Tetrachlorobiphenyl acts an estrogen in vitro and in vivo. Mol. Endocrinol. 10, 923–936. Okuda, K., Uenoyama, Y., Fujita, Y., Iga, K., Sakamoto, K., Kimura, T., 1997. Functional oxytocin receptors in bovine granulosa cells. Biol. Reprod. 56, 625–631. Ozisik, G., Achermann, J.C., Jameson, J.L., 2002. The role of SF-1 in adrenal and reproductive functions: insight from naturally occurring mutations in humans. Mol. Genet. Metab. 76, 85–91. Parker, K.L., Rice, D.A., Lala, D.S., Ikeda, Y., Luo, X., Bakke, M., Zhao, L., Frigieri, C., Hanley, N.A., Stallings, N., Schimmer, B.P., 2002. Steroidogenic factor 1: an essential mediator of endocrine development. Recent Prog. Horm. Res. 57, 19–39.
37
Pogrmic-Majkic, K., Fa, S., Dakic, V., Kaisarevic, S., Kovacevic, R., 2010. Upregulation of peripubertal rat Leydig cell steroidogenesis following 24 h in vitro and in vivo exposure to atrazine. Toxicol. Sci. 118, 52–60. Prakash, B.S., Meyer, H.D., Schallenberger, E., Van de Wiel, D.F., 1987. Development of sensitive enzyme immunoassay (EIA) for progesterone determination in unextracted bovine plasma using the second antibody technique. J. Steroid Biochem. 28, 623–627. Roselli, M., Reinhart, K., Imthurn, B., Keller, P.J., Dubey, R.K., 2000. Cellular and biochemical mechanisms by which environmental estrogens influence reproductive function. Hum. Reprod. Update 6, 332–350. Sadowsky, Y., Crawford, P.A., 1998. Developmental and physiologic roles of the nuclear receptor Steroidogenic Factor-I in the reproductive system. J. Soc. Gynecol. Investig. 5, 6–12. Safe, S.H., Pallaroni, L., Yoon, K., Gaido, K., Ross, S., Saville, B., McDonnell, D., 2001. Toxicology of environmental estrogens. Reprod. Fertil. Dev. 13, 307–315. Saxena, D., Escamilla-Hernandez, R., Little-Ihring, L., Zeleznik, A.J., 2007. Liver receptor homolog-1 and steroidogenic factor-1 have similar actions on rat granulosa cell steroidogenesis. Endocrinology 148, 726–734. Shen, W.H., Moore, C.C.D., Ikeda, Y., Parker, K.L., Ingraham, H.A., 1994. Nuclear receptor steroidogenic factor 1 regulates the Müllerian inhibiting substance gene: a link to the sex determination cascade. Cell 77, 651–661. Tallam, S.K., Walton, J.S., Johnson, W.H., 2000. Effects of oxytocin on follicular development and duration of the estrous cycle in heifers. Theriogenology 53, 951–962. Viger, R.S., Guittot, S.M., Anttonen, M., Wilson, D.B., Heikinheimo, M., 2008. Role of the GATA family of transcription factors in endocrine development, function, and disease. Mol. Endocrinol. 22, 781–798. Voss, A.K., Fortune, J.E., 1992. Oxytocin/neurophysin-I messenger ribonucleic acid in bovine granulosa cells increases after the luteinizing hormone (LH) surge and is stimulated by LH in vitro. Endocrinology 131, 2755–2762. Walther, N., Jansen, M., Akbary, W., Ivell, R., 2006. Differentiation-specific action of orphan nuclear receptor NR5A1 (SF-1): transcriptional regulation in luteinizing bovine theca cells. Reprod. Biol. Endocrinol. 4, 64. Wehrenberg, U., Ivell, R., Jansen, M., von Goedecke, S., Walther, N., 1994. Two orphan receptors binding to a common site are involved in the regulation of the oxytocin gene in the bovine ovary. Proc. Natl. Acad. Sci. U. S. A. 91, 1440–1444. Wilson., V.S., Blystone, C.R., Hotchkiss, A.K., Rider, C.V., Gray Jr., L.E., 2008. Diverse mechanisms of anti-androgen action: impact on male rat reproductive tract development. Int. J. Androl. 31, 178–187. Wojtowicz, A., Tomanek, M., Augustowska, K., Gregoraszczuk, E.L., 2005. Aromatic hydrocarbon receptor (AhR) in the porcine theca and granulosa cells: effect of TCDD, PCB 126 and PCB 153 on the expression of AhR. Endocrine Reg. 39, 109–118. Wrobel, M., Mlynarczuk, J., Kotwica, J., 2009. The adverse effect of dihydrodiphenyltrichloroetane (DDT) and its metabolite (DDE) on the secrertion of prostaglandins and oxytocin in bovine cultured ovarian and endometrial cells. Reprod. Toxicol. 27, 72–78. Zhang, P., Mellon, S.H., 1996. The orphan nuclear receptor steroidogenic factor-1 regulates the cAMP mediated transcriptional activation of rat cytochrome P450c14 (17␣-hydroxylase/c17-20 lyase). Mol. Endocrinol. 10, 147–158. Zhao, L., Bakke, M., Krimkevich, Y., Cushman, L.J., Parlow, A.F., Camper, S.A., Parker, K.L., 2001. Steroidogenic factor 1 (SF1) is essential for pituitary gonadotrope function. Development 128, 147–154.
