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Department of Animal Science, Oklahoma State University, Stillwater, Oklahoma 74078, USA
1 Department of Obstetrics and Gynecology, Stanford University School of Medicine, Stanford, California 94305-5317, USA
(Requests for offprints should be addressed to L J Spicer; Email: igf1Leo{at}okstate.edu)
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Abstract |
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Introduction |
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GDF-9 is most closely related to GDF-9B/BMP-15, both having homology closer to BMPs than to the activin and TGF-ß proteins (Newfeld et al. 1999, Vitt et al. 2002). Members of the TGF-ß family initiate signaling by assembling type I and type II serine/threonine kinase receptor complexes that activate one or more of the eight specific similar to mothers against decapentaplegic (Smad) transcription factors (Massague 1998, Mazerbourg & Hsueh 2003). The type I receptors are also designated as activin receptor-like kinases (ALKs) and are responsible, in part, for transmitting ligand specificity within target cells (Massague 1998, Lux et al. 1999). GDF-9 interacts with the BMP type II receptor (BMPRII) and the type I receptor ALK5 and activates Smad2/3 (Vitt et al. 2002, Kaivo-Oja et al. 2003, Roh et al. 2003, Mazerbourg et al. 2004), whereas BMP-2 and BMP-4 interact with BMPRII and the type I receptors, ALK3 and ALK6 which activate Smad1/5/8 (Massague 1998, Kawabata et al. 1998). However, little is know about the intracellular signaling system GDF-9 and BMP-4 in single-ovulating species such as cattle or humans. Thus, our second objective was to determine the intracellular signaling pathway used by GDF-9 and BMP-4 in bovine granulosa cells.
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Materials and Methods |
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Ovaries from non-pregnant beef cows were collected from a local slaughterhouse and, based on surface diameter, follicular fluid from small (1–5 mm) and large follicles (8–22 mm) was aspirated using 20 gauge needles and 3 ml syringes and centrifuged at 200 g for 5 min to isolate granulosa cells as previously described (Langhout et al. 1991, Spicer & Chamberlain 1998, Spicer et al. 2002). This size classification was based on previous studies showing that granulosa cells from small follicles are less responsive to follicle-stimulating hormone (FSH) and IGF-I than are cells from large follicles (Spicer & Chamberlain 1998, Spicer et al. 2002), follicles 8 mm and greater have much larger estradiol concentrations than small follicles (Stewart et al. 1996, Spicer et al. 2001), and bovine follicles destined to ovulate average 10 ± 2 mm surface diameter (Marion et al. 1968). Granulosa cells were re-suspended in serum-free medium containing collagenase and DNase (Sigma Chemical Co) at 1.25 mg/ml and 0.5 mg/ml respectively, to prevent cell clumping prior to plating. Cells were maintained in this collagenase–DNase-containing medium for less than 1 h prior to dispersion in 1 ml 10% fetal calf serum (FCS) medium without proteases.
Approximately 2.0 x 105 viable cells (in 20–50 µl collagenase–DNase medium) were plated on 24-well Falcon multiwell plates (Becton Dickinson, Lincoln Park, NJ, USA) in a mixture of 1:1 Dulbecco’s modified Eagle’s medium and Ham’s F-12 containing 10% FCS, 0.12 mM gentamycin, 2.0 mM glutamine, and 38.5 mM sodium bicarbonate (all obtained from Sigma Chemical Co) as previously described (Langhout et al. 1991, Spicer & Chamberlain 1998, Spicer et al. 2002). Viability of granulosa cells from small and large follicles was determined by the trypan blue exclusion method, and averaged 58 ± 11% and 79 ± 12% respectively. Cells were cultured in an environment of 95% air and 5% CO2 at 38.5 °C in 10% FCS for the first 48 h with a medium change at 24 h. Cells were then washed twice with serum-free medium and the various treatments (see below) applied in serum-free medium for 48 h with a medium change at 24 h. After the second 24-h treatment period, medium was collected for steroid RIA and cells were collected for cell enumeration (see below).
The hormones used in the cell culture were: FSH (ovine F1913; FSH activity, 15 x NIH-FSH-S1 U/mg) from Scripps Laboratories (San Diego, CA, USA), recombinant human IGF-I and BMP-4 from R&D Systems (Minneapolis, MN, USA), and recombinant rat GDF-9 was generated and characterized as previously described (Hayashi et al. 1999, Vitt et al. 2000a). Briefly, expression vectors for wild-type and epitope-tagged GDF-9 were constructed using pcDNA3.1 Zeo (Invitrogen). N-tagged GDF-9 encoded a Flag epitope (DYKDDDDK) for the M1 antibody followed by six histidine residues fused to the amino terminus of mature GDF-9. Clonal human embryonic kidney 293T cell lines stably expressing wild-type and tagged GDF-9 were used. Quantitation of N-tagged GDF-9 was done after purification with a nickel column and measurement of protein content using Micro BCA protein assay kit (Perstorp Life Science, Rockford, IL, USA). Purified N-tagged GDF-9 was then used as a standard for the quantitation of wild-type GDF-9 in the conditioned medium (serum free) of 293T cells by immunoblots using specific GDF-9 antibodies. In experiments 1, 2, 3, and 5 this GDF-9 preparation was added at 2 µl or less per ml serum-free medium (depending on dose tested). Previous studies (Vitt et al. 2000a, Roh et al. 2003) have shown that conditioned medium from non-transfected human embryonic kidney 293T cells as well as medium with an inactive recombinant N-Tag GDF-9 has no effect on several granulosa cell functions evaluated.
Experimental design
Experiments 1 and 2 were designed to evaluate the dose–response effect of GDF-9 on proliferation and steroidogenesis of small- and large-follicle granulosa cells respectively. Cells were cultured for 48 h in 10% FCS, washed twice with serum-free medium as described earlier, and 0, 150, 300, and 600 ng/ml GDF-9 were applied for 48 h in the presence of either FSH (30 ng/ml) or FSH plus IGF-I (30 ng/ml). The doses of FSH and IGF-I were selected based on previous studies indicating that these doses are maximal for stimulation of estradiol secretion and/or cell proliferation (Spicer & Francisco 1997, Spicer & Chamberlain 1998, Spicer et al. 2002). The doses of GDF-9 were selected based on previous studies (Vitt et al. 2000a, 2002, McNatty et al. 2005a, 2005b).
Experiment 3 was designed to evaluate the effect of GDF-9 on FCS-induced cell proliferation of large-follicle granulosa cells. Cells were cultured for 48 h in 10% FCS, washed twice with serum-free medium as described earlier, and 0, 1%, and 2% of FCS was applied for 48 h in the absence or presence of GDF-9 (300 ng/ml). The dose of GDF-9 was selected based on the results of experiments 1 and 2.
Experiment 4 was designed to evaluate the effect of BMP-4 on proliferation and steroidogenesis of small- and large-follicle granulosa cells respectively. Cells were cultured for 48 h in 10% FCS, washed twice with serum-free medium as described earlier, and 0 and 30 ng/ml BMP-4 were applied for 48 h in the presence of FSH plus IGF-I (30 ng/ml). The dose of BMP-4 was selected based on previous studies indicating that this dose is maximal for its effect on steroidogenesis (Shimasaki et al. 1999, Mulsant et al. 2001, Fabre et al. 2003, Sudo et al. 2004).
Experiment 5 was designed to elucidate the specificity of the GDF-9 and BMP-4 signaling pathway and, based on findings using rat granulosa cells, we tested the ability of GDF-9 and BMP-4 to stimulate the Smad binding element (CAGA) and the BMP response element (BRE) promoter respectively in bovine granulosa cells. The CAGA and BRE promoters are known to be activated by GDF-9 mediated by Smad3 and several BMPs mediated by Smad1/5/8 respectively (Dennler et al. 1998, Kusanagi et al. 2000, Korchynskyi & ten Dijke 2002, Mazerbourg et al. 2004, Monteiro et al. 2004). Granulosa cells from small follicles were cultured as described in experiment 1 with the following treatments applied for 24 h in medium (containing 1% FCS, 30 ng/ml IGF-I, and FSH) after a 4-h transfection with the hormone-specific responsive promoters CAGA or BRE: control (no additions), GDF-9 (300 ng/ml), or BMP-4 (200 ng/ml). The doses of hormones were selected based on the results of experiment 1 and previous in vitro studies (Vitt et al. 2000a, 2002, Mazerbourg et al. 2004) indicating that these doses are effective in altering steroidogenesis and/or promoter activity in granulosa cells.
Experiment 6 was designed to evaluate the effect of purified recombinant human GDF-9 (rhGDF-9; PeproTech Inc, Rock Hill, NJ, USA) and conditioned medium (serum free) from non-transfected human embryonic kidney 293T cells on proliferation and steroidogenesis of small-follicle granulosa cells. Cells were cultured for 48 h in 10% FCS, washed twice with serum-free medium as described earlier, and 0 and 600 ng/ml rhGDF-9, 2 µl conditioned medium from non-transfected 293T cells, and/or 0 or 30 ng/ml FSH were applied for 48 h in the presence of IGF-I (30 ng/ml).
Transfection of granulosa cells
Granulosa cells (2 x 105 viable cells/well) were cultured in 24-well plates in medium supplemented with 10% FCS for 2 days as described earlier. After medium change, cells were incubated in the serum-free medium and transfected with 250 ng DNA/well using Lipofectamine 2000 (Invitrogen) as previously described (Mazerbourg et al. 2004). Briefly, the pCMV-ß-galactosidase plasmid (50 ng) was co-transfected to monitor transfection efficiency. After transfection, cells were treated with GDF-9 or BMP-4 for 24 h in medium containing 1% FCS. To harvest cells, lysis buffer (200 µl; Promega Corp) was added into each well and 30 µl of the supernatant was used for luciferase determination using a luminometer (Luminark microplate reader; Bio-Rad Laboratories, Inc.). Fifty microliters of the cell lysate were also used to measure the ß-galactosidase activity to monitor transfection efficiency. The reporter activity is expressed as the ratio of relative light units/ß-galactosidase activity.
Determination of cell numbers and steroid concentrations
Medium was collected from individual wells and frozen at –20 °C for subsequent hormone analyses. Cells were then gently washed twice with 0.9% saline (500 µl), exposed to 500 µl trypsin solution (0.25%, w/v) for 20 min at 25 °C, and then scraped from each well. Cell aggregates were minimized by pipetting cell suspensions back and forth through a 500 µl pipette tip three to five times. Cells were then diluted in 9 ml 0.9% saline, and counted using a Coulter counter (model Zm; Coulter Electronics, Hialeah, FL, USA) as previously described (Langhout et al. 1991, Spicer & Chamberlain 1998, Spicer et al. 2002).
Concentrations of progesterone and estradiol in culture medium were determined by RIA as previously described (Langhout et al. 1991, Spicer et al. 2002). The intra- and interassay coefficients of variation were 12% and 17% for the progesterone RIA, and 7% and 14% for the estradiol RIA.
Statistical analysis
Each experiment contained three replicates per treatment and each experiment was replicated three to four times with different pools of granulosa cells. Each pool of small-follicle granulosa cells was obtained from multiple follicles collected from seven to ten cattle, and each pool of large-follicle granulosa cells was obtained from four to seven follicles collected from three to six cattle. Data are presented as the least squares means ( ± S.E.) of measurements from nine to twelve culture wells. Main effects (i.e. hormone, dose, experimental replicate) and interactions were assessed using the general linear model procedure of SAS (1999). Steroid production was expressed as ng or pg/105 cells per 24 h, and cell numbers at the termination of the experiment were used for this calculation. Specific differences in cell numbers, steroid production, and promoter (luciferase) activity among treatments were determined using Fisher’s protected least significant difference procedure (Ott 1977).
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Cell numbers
GDF-9 caused a dose-dependent increase (P<0.05) in basal (60% increase) and IGF-I-induced (28% increase) cell numbers (Fig. 1A
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). At 150 ng/ml, GDF-9 had no significant effect on progesterone production (Fig. 2A). In the absence of IGF-I, only 600 ng/ml decreased (by 28%; P<0.05) progesterone production in FSH-treated cultures (Fig. 2A).
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). At 300 ng/ml, GDF-9 inhibited (P<0.05) estradiol production by 27%. In the absence of IGF-I, none of the doses of GDF-9 affected (P>0.10) estradiol production in FSH-treated cultures (Fig. 2B). Experiment 2: GDF-9 effects on large-follicle granulosa cells
Cell numbers
GDF-9 caused a dose-dependent increase (P<0.01) in basal (maximum 2.1-fold increase) and IGF-I-induced (maximum 2.3-fold increase) cell numbers (Fig. 1B).
Progesterone production
IGF-I increased (P<0.001) progesterone production (to 2-fold of FSH-treated controls) by granulosa cells, and GDF-9 caused a dose-dependent inhibition (P<0.05) of this increase with 600 ng/ml completely blocking the IGF-I-induced increase (Fig. 3A). In contrast to small-follicle granulosa cells, 150 ng/ml GDF-9 significantly decreased progesterone production (Fig. 3A). In the absence of IGF-I but in the presence of FSH, only 600 ng/ml decreased (by 38%; P<0.05) progesterone production (Fig. 3A).
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). In the absence of IGF-I but in the presence of FSH, none of the doses of GDF-9 affected (P>0.10) estradiol production (Fig. 3B). Experiment 3: Effect of GDF-9 on FCS-induced large-follicle granulosa cell growth
Cell numbers
GDF-9 amplified (P<0.05) basal and FCS-induced cell numbers (Fig. 4). Alone, GDF-9 increased (P<0.05) cells numbers by 2.1-fold whereas 1% and 2% FCS increased (P<0.05) cell numbers by 4.7- and 5.9-fold respectively (Fig. 4). In the presence of 1% and 2% FCS, GDF-9 further increased (P<0.05) cell numbers by 31% and 27% respectively (Fig. 4).
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Cell numbers BMP-4 had no effect (P>0.10) on numbers of granulosa cells collected from small (control= 1.37 and BMP-4=1.54 ± 0.04 x 105 cells/well) or large (control=1.08 and BMP-4=1.22 ± 0.06 x 105 cells/well) follicles.
Progesterone production
BMP-4 inhibited (P<0.05) progesterone production induced by IGF-I and FSH by 39% and 27% in cultures of small- and large-follicle granulosa cells respectively (Fig. 5A).
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). Experiment 5: GDF-9 and BMP-4 effects on CAGA and BRE promoter activity in small-follicle granulosa cells
Granulosa cells from small follicles were cultured for 48 h in 10% FCS, followed by transfection with either the CAGA or BRE promoter and hormonal treatments revealed that treatment with GDF-9 increased (P<0.01) CAGA promoter activity 1.8-fold of controls, whereas BMP-4 was without effect (P>0.10; Fig. 6A). In contrast, BMP-4 increased (P<0.01) BRE promoter activity by 3.9-fold of controls, whereas GDF-9 was without effect (P>0.10; Fig. 6B).
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Cell numbers
rhGDF-9 but not FSH or conditioned medium from 293T cells increased (P<0.05) the numbers of granulosa cells collected from small follicles (Table 1).
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). Conditioned medium from 293T cells had no significant effect on progesterone production (Table 1).
Estradiol production
FSH increased (P<0.05) estradiol production (to 4.7-fold of IGF-I-treated controls) by granulosa cells, and rhGDF-9 significantly reduced this FSH-induced increase by 40% (Table 1). Conditioned medium from 293T cells had no significant effect on estradiol production (Table 1).
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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In the present study, stimulatory effects of GDF-9 were observed on basal and IGF-I-induced granulosa cell proliferation whether cells were obtained from large or small follicles. These observations agree with previous reports in which treatment with GDF-9 (10–2000 ng/ml) increased numbers of rat granulosa cells (by over 2-fold) as well as cell proliferation (by 2- to 9-fold) as measured by [3H]thymidine incorporation (Vitt et al. 2000a, 2002, McNatty et al. 2005a). In contrast, a recent report indicates that treatment with either 1 or 2 µg/ml recombinant ovine GDF-9 had no significant effect on [3H]thymidine incorporation into ovine or bovine granulosa cells (McNatty et al. 2005b). In further agreement with the present study, the proliferative responses of rat granulosa cells to GDF-9 observed by Vitt et al. (2000a) was qualitatively similar whether cells were obtained from small antral follicles or large preovulatory follicles, although the maximally effective dose of GDF-9 was 5-fold lower in rat granulosa cells obtained from large preovulatory follicles. The latter suggests that rat granulosa cells of mature differentiated follicles may be more sensitive to GDF-9 than those of immature undifferentiated follicles. Data in the present study support this assumption because large-follicle granulosa cell proliferation and progesterone production (see next paragraph) were significantly affected by the lowest dose of GDF-9 tested, whereas small-follicle granulosa cells were not. Interestingly, BMP-4 at a dose effective in inhibiting steroidogenesis had no effect on IGF-I-induced cell numbers in the present study. Shepherd & Nachtigal (2003) reported that neither 2- nor 4-day BMP-4 treatment affected FCS-induced proliferation of four types of human ovarian carcinoma cells. Also, others have reported that BMP-4 (Fabre et al. 2003) and BMP-2 (Souza et al. 2002) have no effect on ovine granulosa cell proliferation while significantly affecting steroidogenesis in vitro. Thus, the present study supports the notion that two TGF-ß superfamily members that bind to the same BMPRII receptor (Kawabata et al. 1998, Lux et al. 1999, Newfeld et al. 1999) but derived from two different (i.e. oocyte vs theca) cell types have diverse biological responses within the granulosa layer, the latter of which are mediated by the various type I receptors (ALKs) and intracellular Smad response proteins (Massague 1998, Mazerbourg & Hsueh 2003, Mazerbourg et al. 2004). Whether GDF-9 and BMP-4 have stimulatory effects on preantral follicular growth in cattle as they do in rodents (Vitt et al. 2000b, Nilsson & Skinner 2002, 2003, Latham et al. 2004, Wang & Roy 2004) and humans (Hreinsson et al. 2002) awaits further elucidation. Also, whether GDF-9 plays a role in the regulation of ovulation rate in cattle, as recently demonstrated for sheep (Hanrahan et al. 2004, Juengel et al. 2004), will require additional research.
Previous studies with rat granulosa cells have reported that 48-h (Vitt et al. 2000a, 2002) and 6-day (McNatty et al. 2005a) treatment with>30 ng/ml GDF-9 inhibits progesterone production induced by FSH. Similarly, in the present study, GDF-9 decreased bovine granulosa cell progesterone production in the presence of FSH alone (highest dose of GDF-9 only) or a combination of FSH and IGF-I (all doses of GDF-9). McNatty et al. (2005b) recently reported that 6-day treatment with 2 µg/ml GDF-9 inhibited progesterone production by ovine and bovine granulosa cells cultured in the presence of FSH, IGF-I, and insulin. Yamamoto et al.(2002) also observed that GDF-9 inhibits progesterone production induced by 8-bromo-cAMP in cultured human granulosa cells. In the present study, under basal conditions (the presence of FSH), GDF-9 (only 600 ng/ml) had a weak inhibitory effect on progesterone production. In previous studies with cultured mouse granulosa cells, 50–100 ng/ml GDF-9 treatment for 16–24 h increased basal progesterone production but had no effect on FSH-induced progesterone production (Elvin et al. 1999, 2000). Discrepancies among studies could be due to species differences and/or differences in culture conditions (e.g. duration of treatment or presence or absence of FCS). Consistent with previous reports (Spicer & Chamberlain 1998, Spicer et al. 2002), IGF-I stimulated progesterone and estradiol production by bovine granulosa cells in the presence of FSH (Figs 2 and 3). It should be emphasized that granulosa cells in the present and previous studies, because of serum exposure (Orly et al. 1980, Luck et al. 1990), may have partially luteinized and therefore be exhibiting some luteal cell activity. The less than 5-fold increase in estradiol secretion induced by a high dose (i.e. 30 ng/ml) of FSH combined with the very high ratio (i.e.>150) of progesterone:estradiol secreted by the granulosa cells of the present study supports this latter suggestion.
Reported for the first time using bovine granulosa cells, GDF-9 inhibited estradiol secretion by granulosa cells obtained from large and small follicles of a single-ovulating species (Figs 2B and 3B). Previously, treatment of rat granulosa cells with GDF-9 (30–150 ng/ml) inhibited (by up to 58%) FSH-induced estradiol production without affecting forskolin-induced estradiol production (Vitt et al. 2000a). Thus, granulosa cell aromatase activity from multiple- and single-ovulating species appear to respond similarly to GDF-9. Also consistent with the present study, Vitt et al. (2000a) found that the inhibitory effect of GDF-9 on estradiol production was similar whether cells were obtained from small antral or large preovulatory rat follicles. Collectively, these studies indicate that GDF-9 is an effective suppressor of aromatase activity regardless of the size (or differentiation state) of the follicle in which the granulosa cells reside.
Similar to GDF-9, BMP-4 inhibited both progesterone and estradiol production by bovine granulosa cells from small and large follicles. Although originally identified for their ability to induce bone and cartilage formation (Wan & Cao 2005), BMPs have since been shown to play important roles in cellular differentiation and formation in many other tissues and organs including the ovary (Massague 1998, Knight & Glister 2003, Mazerbourg & Hsueh 2003). Inhibitory effects of BMP-4 on progesterone production by granulosa cells of sheep (Mulsant et al. 2001, Fabre et al. 2003, Pierre et al. 2004), cattle (Glister et al. 2004), and rats (Shimasaki et al. 1999, Sudo et al. 2004) have been documented. The present study further evaluated BMP-4 effects on aromatase activity. In cultured rat granulosa cells, BMP-4 stimulates FSH-induced estradiol production (Shimasaki et al. 1999, Sudo et al. 2004) but its effect in the presence of IGF-I was not studied. One other report indicated that a 6-day treatment of bovine granulosa cells with BMP-4 increases estradiol production and cell proliferation (Glister et al. 2004) but because estradiol production was not corrected for increased cell numbers, clear interpretation of these data is not possible. In the present study, BMP-4 suppressed FSH plus IGF-I-induced aromatase activity as well as progesterone production in bovine granulosa cells regardless of whether cells originated from small or large follicles. More extensive studies are needed to determine if responsiveness to BMP-4 changes during follicular development and if locally produced BMPs change during follicular development in cattle.
Using bovine granulosa cells, we have shown that GDF-9 (but not BMP-4) activated the Smad3-induced CAGA promoter, whereas BMP-4 (but not GDF-9) activated the Smad1/5/8-induced BRE promoter. Similar results has been obtained in rat and human granulosa cells (Kaivo-Oja et al. 2003, 2005, Roh et al. 2003, Mazerbourg et al. 2004, Sudo et al. 2004). Previous studies in rat and human granulosa cells have suggested that GDF-9 induces the phosphorylation of Smad2/3 after interaction with the receptor type II, BMPRII, and the receptor type I, ALK5 (Vitt et al. 2002, Mazerbourg et al. 2004, Kaivo-Oja et al. 2005). Stimulation of the CAGA promoter is mediated by TGF-ß, activin, or GDF-9 after phosphorylation of the downstream Smad3 proteins, whereas BMP-2 and BMP-7 activate Smad1, 5, and 8 (Massague 1998) and subsequently the BRE and GCCG promoters (Kusanagi et al. 2000, Korchynskyi & ten Dijke 2002). However, the distinct biological effects of the TGF-ß superfamily members may be exerted by the transcriptional regulation of target genes in a cell-type specific manner that may involve ALK-specific and opposing effects that may include the inhibitory Smads (Smad6 and Smad7) (ten Dijke & Hill 2004) and may explain how factors such as GDF-9 have both stimulatory and inhibitory effects on granulosa cell functions. Further studies will be required to elucidate the elements involved in the inhibitory (steroidogenesis) versus stimulatory (cell proliferation) action of GDF-9.
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References |
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Carabatsos MJ, Elvin J, Matzuk MM & Albertini DF 1998 Characterization of oocyte and follicle development in growth differentiation factor-9-deficient mice. Developmental Biology 204 373–384.[CrossRef][ISI][Medline]
Dennler S, Itoh S, Vivien D, ten Dijke P, Huet S & Gauthier JM 1998 Direct binding of Smad3 and Smad4 to critical TGF ß-inducible elements in the promoter of human plasminogen activator inhibitor-type 1 gene. EMBO Journal 17 3091–3100.[CrossRef][ISI][Medline]
Dong J, Albertini DF, Nishimori K, Kumar TR, Lu N & Matzuk MM 1996 Growth differentiation factor-9 is required during early ovarian folliculogenesis. Nature 383 531–535.[CrossRef][Medline]
Elvin JA, Clark AT, Wang P, Wolfman NM & Matzuk MM 1999 Paracrine actions of growth differentiation factor-9 in the mammalian ovary. Molecular Endocrinology 13 1035–1048.
Elvin JA, Yan C & Matzuk MM 2000 Growth differentiation factor-9 stimulates progesterone synthesis in granulosa cells via a prostaglandin E2/EP2 receptor pathway. PNAS 97 10288–10293.
Erickson GF & Shimasaki S 2003 The spatiotemporal expression pattern of the bone morphogenetic protein family in rat ovary cell types during the estrous cycle. Reproductive Biology and Endocrinology 1 9–29.[CrossRef]
Fabre S, Pierre A, Pisselet C, Mulsant P, Lecerf F, Pohl J, Monget P & Monniaux D 2003 The Booroola mutation in sheep is associated with an alteration of the bone morphogenetic protein receptor-IB functionality. Journal of Endocrinology 177 435–444.[Abstract]
Fatehi AN, van den Hurk R, Colenbrander B, Daemen AJ, van Tol HT, Monteiro RM, Roelen BA & Bevers MM 2005 Expression of bone morphogenetic protein2 (BMP2), BMP4 and BMP receptors in the bovine ovary but absence of effects of BMP2 and BMP4 during IVM on bovine oocyte nuclear maturation and subsequent embryo development. Theriogenology 63 872–889.[CrossRef][ISI][Medline]
Glister C, Kemp CF & Knight PG 2004 Bone morphogenetic protein (BMP) ligands and receptors in bovine ovarian follicle cells: actions of BMP-4, -6 and -7 on granulosa cells and differential modulation of Smad-1 phosphorylation by follistatin. Reproduction 127 239–254.
Hanrahan JP, Gregan SM, Mulsant P, Mullen M, Davis GH, Powell R & Galloway SM 2004 Mutations in the genes for oocyte-derived growth factors GDF9 and BMP15 are associated with both increased ovulation rate and sterility in Cambridge and Belclare sheep (Ovis aries). Biology of Reproduction 70 900–909.
Hayashi M, McGee EA, Min G, Klein C, Rose UM, van Duin M & Hsueh AJ 1999 Recombinant growth differentiation factor-9 (GDF-9) enhances growth and differentiation of cultured early ovarian follicles. Endocrinology 140 1236–1244.
Hreinsson JG, Scott JE, Rasmussen C, Swahn ML, Hsueh AJ & Hovatta O 2002 Growth differentiation factor-9 promotes the growth, development, and survival of human ovarian follicles in organ culture. Journal of Clinical Endocrinology and Metabolism 87 316–321.
Hunter MG, Robinson RS, Mann GE & Webb R 2004 Endocrine and paracrine control of follicular development and ovulation rate in farm species. Animal Reproduction Science 82–83 461–477.
Jaatinen R, Laitinen MP, Vuojolainen K, Aaltonen J, Louhio H, Heikinheimo K, Lehtonen E & Ritvos O 1999 Localization of growth differentiation factor-9 (GDF-9) mRNA and protein in rat ovaries and cDNA cloning of rat GDF-9 and its novel homolog GDF-9B. Molecular and Cellular Endocrinology 156 189–193.[CrossRef][ISI][Medline]
Juengel JL & McNatty KP 2005 The role of proteins of the transforming growth factor-ß superfamily in the intraovarian regulation of follicular development. Human Reproduction Update 11 143–160.[Medline]
Juengel JL, Hudson NL, Heath DA, Smith P, Reader KL, Lawrence SB, O’Connell AR, Laitinen MP, Cranfield M, Groome NP et al. 2002 Growth differentiation factor 9 and bone morphogenetic protein 15 are essential for ovarian follicular development in sheep. Biology of Reproduction 67 1777–1789.
Juengel JL, Hudson NL, Whiting L & McNatty KP 2004 Effects of immunization against bone morphogenetic protein 15 and growth differentiation factor 9 on ovulation rate, fertilization, and pregnancy in ewes. Biology of Reproduction 70 557–561.
Kaivo-Oja N, Bondestam J, Kamarainen M, Koskimies J, Vitt U, Cranfield M, Vuojolainen K, Kallio JP, Olkkonen VM, Hayashi M et al. 2003 Growth differentiation factor-9 induces Smad2 activation and inhibin B production in cultured human granulosa-luteal cells. Journal of Clinical Endocrinology and Metabolism 88 755–762.
Kaivo-Oja N, Mottershead DG, Mazerbourg S, Myllymaa S, Duprat S, Gilchrist RB, Groome NP, Hsueh AJ & Ritvos O 2005 Adenoviral gene transfer allows Smad-responsive gene promoter analyses and delineation of type I receptor usage of transforming growth factor-ß family ligands in cultured human granulosa luteal cells. Journal of Clinical Endocrinology and Metabolism 90 271–278.
Kawabata M, Imamura T & Miyazono K 1998 Signal transduction by bone morphogenetic proteins. Cytokine and Growth Factor Reviews 9 49–61.[CrossRef][ISI][Medline]
Knight PG & Glister C 2003 Local roles of TGF-ß superfamily members in the control of ovarian follicle development. Animal Reproduction Science 78 165–183.[CrossRef][ISI][Medline]
Korchynskyi O & ten Dijke P 2002 Identification and functional characterization of distinct critically important bone morphogenetic protein-specific response elements in the Id1 promoter. Journal of Biological Chemistry 277 4883–4891.
Kusanagi K, Inoue H, Ishidou Y, Mishima HK, Kawabata M & Miyazono K 2000 Characterization of a bone morphogenetic protein-responsive Smad-binding element. Molecular Biology of the Cell 11 555–565.
Langhout DJ, Spicer LJ & Geisert RD 1991 Development of a culture system for bovine granulosa cells: effects of growth hormone, estradiol, and gonadotropins on cell proliferation, steroidogenesis, and protein synthesis. Journal of Animal Science 69 3321–3334.[Abstract]
Latham KE, Wigglesworth K, McMenamin M & Eppig JJ 2004 Stage-dependent effects of oocytes and growth differentiation factor 9 on mouse granulosa cell development: advance programming and subsequent control of the transition from preantral secondary follicles to early antral tertiary follicles. Biology of Reproduction 70 1253–1262.
Lonergan P, Gutierrez-Adan A, Rizos D, Pintado B, de la Fuente J & Boland MP 2003 Relative messenger RNA abundance in bovine oocytes collected in vitro or in vivo before and 20 hr after the preovulatory luteinizing hormone surge. Molecular Reproduction and Development 66 297–305.[CrossRef][ISI][Medline]
Luck MR, Rodgers RJ & Findlay JK 1990 Secretion and gene expression of inhibin, oxytocin and steroid hormones during the in vitro differentiation of bovine granulosa cells. Reproduction, Fertility and Development 2 11–25.[CrossRef][Medline]
Lux A, Attisano L & Marchuk DA 1999 Assignment of transforming growth factor ß1 and ß3 and a third new ligand to the type I receptor ALK-1. Journal of Biological Chemistry 274 9984–9992.
Marion GB, Gier HT & Choudary JB 1968 Micromorphology of the bovine ovarian follicular system. Journal of Animal Science 27 451–465.
Massague J 1998 TGF-ß signal transduction. Annual Reviews of Biochemistry 67 753–791.
Mazerbourg S & Hsueh AJ 2003 Growth differentiation factor-9 signaling in the ovary. Molecular and Cellular Endocrinology 202 31–36.[ISI][Medline]
Mazerbourg S, Bondy CA, Zhou J & Monget P 2003 The insulin-like growth factor system: a key determinant role in the growth and selection of ovarian follicles? A comparative species study. Reproduction in Domestic Animals 38 247–258.[CrossRef][ISI][Medline]
Mazerbourg S, Klein C, Roh J, Kaivo-Oja N, Mottershead DG, Korchynskyi O, Ritvos O & Hsueh AJ 2004 Growth differentiation factor-9 signaling is mediated by the type I receptor, activin receptor-like kinase 5. Molecular Endocrinology 18 653–665.
McGee EA & Hsueh AJ 2000 Initial and cyclic recruitment of ovarian follicles. Endocrine Reviews 21 200–214.
McGrath SA, Esquela AF & Lee SJ 1995 Oocyte-specific expression of growth/differentiation factor-9. Molecular Endocrinology 9 131–136.[Abstract]
McNatty KP, Juengel JL, Reader KL, Lun S, Myllymaa S, Lawrence SB, Western A, Meerasahib MF, Mottershead DG, Groome NP et al. 2005a Bone morphogenetic protein 15 and growth differentiation factor 9 co-operate to regulate granulosa cell function. Reproduction 129 473–480.
McNatty KP, Juengel JL, Reader KL, Lun S, Myllymaa S, Lawrence SB, Western A, Meerasahib MF, Mottershead DG, Groome NP et al. 2005b Bone morphogenetic protein 15 and growth differentiation factor 9 co-operate to regulate granulosa cell function in ruminants. Reproduction 129 481–487.
Monteiro RM, de Sousa Lopes SM, Korchynskyi O, ten Dijke P & Mummery CL 2004 Spatio-temporal activation of Smad1 and Smad5 in vivo: monitoring transcriptional activity of Smad proteins. Journal of Cell Science 117 4653–4663.
Mulsant P, Lecerf F, Fabre S, Schibler L, Monget P, Lanneluc I, Pisselet C, Riquet J, Monniaux D, Callebaut I et al. 2001 Mutation in bone morphogenetic protein receptor-IB is associated with increased ovulation rate in Booroola Merino ewes. PNAS 98 5104–5109.
Newfeld SJ, Wisotzkey RG & Kumar S 1999 Molecular evolution of a developmental pathway: phylogenetic analyses of transforming growth factor-ß family ligands, receptors and Smad signal transducers. Genetics 152 783–795.
Nilsson EE & Skinner MK 2002 Growth and differentiation factor-9 stimulates progression of early primary but not primordial rat ovarian follicle development. Biology of Reproduction 67 1018–1024.
Nilsson EE & Skinner MK 2003 Bone morphogenetic protein-4 acts as an ovarian follicle survival factor and promotes primordial follicle development. Biology of Reproduction 69 1265–1272.
Orly J, Sato G & Erickson GF 1980 Serum suppresses the expression of hormonally induced functions in cultured granulosa cells. Cell 20 817–827.[CrossRef][ISI][Medline]
Ott L 1977 Multiple comparisons. In An Introduction to Statistical Methods and Data Analysis, pp 384–388. North Scituate, MA, USA: Duxbury Press.
Pennetier S, Uzbekova S, Perreau C, Papillier P, Mermillod P & Dalbies-Tran R 2004 Spatio-temporal expression of the germ cell marker genes MATER, ZAR1, GDF9, BMP15, and VASA in adult bovine tissues, oocytes, and preimplantation embryos. Biology of Reproduction 71 1359–1366.
Pierre A, Pisselet C, Dupont J, Mandon-Pepin B, Monniaux D, Monget P & Fabre S 2004 Molecular basis of bone morphogenetic protein-4 inhibitory action on progesterone secretion by ovine granulosa cells. Journal of Molecular Endocrinology 33 805–817.
Prochazka R, Nemcova L, Nagyova E & Kanka J 2004 Expression of growth differentiation factor 9 messenger RNA in porcine growing and preovulatory ovarian follicles. Biology of Reproduction 71 1290–1295.
Roh JS, Bondestam J, Mazerbourg S, Kaivo-Oja N, Groome N, Ritvos O & Hsueh AJ 2003 Growth differentiation factor-9 stimulates inhibin production and activates Smad2 in cultured rat granulosa cells. Endocrinology 144 172–178.
SAS 1999 The SAS System for Windows, version 8. Cary, NC, USA: SAS Institute Inc.
Sendai Y, Itoh T, Yamashita S & Hoshi H 2001 Molecular cloning of a cDNA encoding a bovine growth differentiation factor-9 (GDF-9) and expression of GDF-9 in bovine ovarian oocytes and in vitro-produced embryos. Cloning 3 3–10.[CrossRef][Medline]
Shepherd TG & Nachtigal MW 2003 Identification of a putative autocrine bone morphogenetic protein-signaling pathway in human ovarian surface epithelium and ovarian cancer cells. Endocrinology 144 3306–3314.
Shimasaki S, Zachow RJ, Li D, Kim H, Iemura S, Ueno N, Sampath K, Chang RJ & Erickson GF 1999 A functional bone morphogenetic protein system in the ovary. PNAS 96 7282–7287.
Souza CJ, Campbell BK, McNeilly AS & Baird DT 2002 Effect of bone morphogenetic protein 2 (BMP2) on oestradiol and inhibin A production by sheep granulosa cells, and localization of BMP receptors in the ovary by immunohistochemistry. Reproduction 123 363–369.[Abstract]
Spicer LJ 2004 Proteolytic degradation of insulin-like growth factor binding proteins by ovarian follicles: a control mechanism for selection of dominant follicles. Biology of Reproduction 70 1223–1230.
Spicer LJ & Francisco CC 1997 The adipose obese gene product, leptin: evidence of a direct inhibitory role in ovarian function. Endocrinology 138 3374–3379.
Spicer LJ & Chamberlain CS 1998 Influence of cortisol on insulin-and insulin-like growth factor 1 (IGF-1)-induced steroid production and on IGF-1 receptors in cultured bovine granulosa cells and thecal cells. Endocrine 9 153–161.[CrossRef][ISI][Medline]
Spicer LJ, Chamberlain CS & Morgan GL 2001 Proteolysis of insulin-like growth factor binding proteins during preovulatory follicular development in cattle. Domestic Animal Endocrinology 21 1–15.[CrossRef][ISI][Medline]
Spicer LJ, Chamberlain CS & Maciel SM 2002 Influence of gonadotropins on insulin- and insulin-like growth factor-I (IGF-I)-induced steroid production by bovine granulosa cells. Domestic Animal Endocrinology 22 237–254.[CrossRef][ISI][Medline]
Stewart RE, Spicer LJ, Hamilton TD, Keefer BE, Dawson LJ, Morgan GL & Echternkamp SE 1996 Levels of insulin-like growth factor (IGF) binding proteins, luteinizing hormone and IGF-I receptors, and steroids in dominant follicles during the first follicular wave in cattle exhibiting regular estrous cycles. Endocrinology 137 2842–2850.[Abstract]
Sudo S, Avsian-Kretchmer O, Wang LS & Hsueh AJ 2004 Protein related to DAN and cerberus is a bone morphogenetic protein antagonist that participates in ovarian paracrine regulation. Journal of Biological Chemistry 279 23134–23141.
ten Dijke P & Hill CS 2004 New insights into TGF-ß-Smad signalling. Trends in Biochemical Sciences 29 265–273.[CrossRef][ISI][Medline]
Vitt UA, Hayashi M, Klein C & Hsueh AJ 2000a Growth differentiation factor-9 stimulates proliferation but suppresses the follicle-stimulating hormone-induced differentiation of cultured granulosa cells from small antral and preovulatory rat follicles. Biology of Reproduction 62 370–377.
Vitt UA, McGee EA, Hayashi M & Hsueh AJ 2000b In vivo treatment with GDF-9 stimulates primordial and primary follicle progression and theca cell marker CYP17 in ovaries of immature rats. Endocrinology 141 3814–3820.
Vitt UA, Mazerbourg S, Klein C & Hsueh AJ 2002 Bone morphogenetic protein receptor type II is a receptor for growth differentiation factor-9. Biology of Reproduction 67 473–480.
Wan M & Cao X 2005 BMP signaling in skeletal development. Biochemical and Biophysical Research Communications 328 651–657.[CrossRef][ISI][Medline]
Wang J & Roy SK 2004 Growth differentiation factor-9 and stem cell factor promote primordial follicle formation in the hamster: modulation by follicle-stimulating hormone. Biology of Reproduction 70 577–585.
Yamamoto N, Christenson LK, McAllister JM & Strauss JF III 2002 Growth differentiation factor-9 inhibits 3'5'-adenosine monophosphate-stimulated steroidogenesis in human granulosa and theca cells. Journal of Clinical Endocrinology and Metabolism 87 2849–2856.
Received in final form 4 February 2006
Accepted 23 February 2006
Made available online as an Accepted Preprint 24 February 2006
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