Journal: Cell Communication and Signaling : CCS

Article Title: GDF9 His209GlnfsTer6/S428T and GDF9 Q321X/S428T bi-allelic variants caused female subfertility with defective follicle enlargement

doi: 10.1186/s12964-024-01616-8

Figure Lengend Snippet: Effects of GDF9 variants on gene expression and GDF9-BMP15 interactions. A Western blots of mature GDF9 proteins expressed in follicular fluid collected from the first and second retrieval cycle of individual P1 and age-matched controls ( n =3). B ELISA of GDF9 in follicular fluid collected from the first retrieval cycle of individual P1 and age-matched controls ( n =6). C Western blots of GDF9 expression in HEK293T cells transfected with human GDF9 WT , GDF9 Q321X , GDF9 S428T , and GDF9 His209GlnfsTer6 respectively and their relative mature-GDF9 expression in culture medium (CM) (3 independent experiments). The amount of concentrated culture medium used for the detection was 35μl per sample per assay. lnfsTer6, His209GlnfsTer6. * P < 0.05. D Western blots of GDF9 precursors and mature GDF9 over time during in vitro cleavage assay (3 independent experiments). Total GDF9=GDF9 precursor+mature GDF9. E Western blots of BMP15 expression in HEK293T cells co-transfected with human BMP15 and human GDF9 WT , GDF9 Q321X , GDF9 S428T , and GDF9 His209GlnfsTer6 respectively and their relative BMP15 precursor expression in CM (3 independent experiments). lnfsTer6, His209GlnfsTer6. * P < 0.05. F Co-IP assay of the binding of GDF9 proteins to BMP15 protein. lnfsTer6, His209GlnfsTer6. * P < 0.05. G Western blots of p-smad2 and smad2 expression in human primary luteinized granulosa cells treated with purified recombinant pro-GDF9 WT -BMP15 and pro-GDF9 S428T -BMP15 protein respectively (3 independent experiments)

Article Snippet: GDF9 was measured in follicular fluid collected at the first oocyte retrieval from individual P1 and in follicular fluid from other six control women using an ELISA kit (CSB-E12925h, CUSABIO, China).

Techniques: Gene Expression, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Transfection, In Vitro, Cleavage Assay, Co-Immunoprecipitation Assay, Binding Assay, Purification, Recombinant

Journal: Endocrinology

Article Title: Investigations of TGF-β signaling in preantral follicles of female mice reveal differential roles for bone morphogenetic protein 15.

doi: 10.1210/en.2012-2251

Figure Lengend Snippet: Figure 1. Individual effects of BMP15 or GDF9 on preantral follicle growth in vitro. Follicles were mechanically isolated and cultured individually in 96-well plates for 72 h in a range of concentrations of BMP15 (a,b) or GDF9 (c,d) and with 10ng/ml FSH (b,d). Variation in follicle size and morphology exposed to different treatments are shown (e). Values are means SEM of 4-6 cultures. Comparisons between treatment groups were made within each time point by ANOVA and Bonferroni multiple comparisons test where *P .05 and **P .01 vs 0ng/ml.

Article Snippet: Follicles were exposed to a range of concentrations of recombinant mouse (rm) GDF9 or recombinant human (rh) BMP15 (R&D Systems Inc., Abingdon, UK).

Techniques: In Vitro, Isolation, Cell Culture

Journal: Endocrinology

Article Title: Investigations of TGF-β signaling in preantral follicles of female mice reveal differential roles for bone morphogenetic protein 15.

doi: 10.1210/en.2012-2251

Figure Lengend Snippet: Figure 2. Combined effects of BMP15 and GDF9 on preantral follicle growth in vitro. Follicles were mechanically isolated and cultured individually in 96-well plates in BMP15 (250ng/ml), GDF9 (250ng/ml) or equal amounts (250ng/ml) of both and overall size relative to control was assessed at each time point (a). The co-operative effect of both ligands on follicle growth was assessed further using a fixed concentration of GDF9 (250ng/ml) and a range of concentrations of BMP15 (0-250ng/ml) (b). Values are means SEM (n 8 (a) and n 5 (b) cultures). Comparisons between treatment groups were made within each time point by ANOVA and Bonferroni multiple comparisons test where *P .05 and **P .01 vs control.

Article Snippet: Follicles were exposed to a range of concentrations of recombinant mouse (rm) GDF9 or recombinant human (rh) BMP15 (R&D Systems Inc., Abingdon, UK).

Techniques: In Vitro, Isolation, Cell Culture, Control, Concentration Assay

Journal: Endocrinology

Article Title: Investigations of TGF-β signaling in preantral follicles of female mice reveal differential roles for bone morphogenetic protein 15.

doi: 10.1210/en.2012-2251

Figure Lengend Snippet: Figure 3. Effect of BMP15 and/or GDF9 on GC proliferation and apoptosis in cultured preantral follicles. Follicles were fixed after 24 or 48 h culture with BMP15 (250ng/ml), GDF9 (250ng/ml) or equal amounts (250ng/ml) of both. Sections of follicles were stained for Ki67 (green) or cleaved caspase-3 (red) and counterstained with DAPI (blue), examples at 24h are shown (a). Positive staining in the GC compartment was quantified for Ki67 and cleaved caspase-3 at 24h (b) and 48h (c). Values are means SEM (n 4-6 follicles/group). *P .05, **P .01 vs control (Cntrl). Scale bar represents 50 m.

Article Snippet: Follicles were exposed to a range of concentrations of recombinant mouse (rm) GDF9 or recombinant human (rh) BMP15 (R&D Systems Inc., Abingdon, UK).

Techniques: Cell Culture, Staining, Control

Journal: Endocrinology

Article Title: Investigations of TGF-β signaling in preantral follicles of female mice reveal differential roles for bone morphogenetic protein 15.

doi: 10.1210/en.2012-2251

Figure Lengend Snippet: Figure 4. Candidate gene expression in cultured preantral follicles exposed to BMP15 and/or GDF9. Preantral follicles were cultured for 24 h in BMP15 (250ng/ml), GDF9 (250ng/ml) or equal amounts (250ng/ml) of both. Samples from each group consist of four pooled follicles that were snap frozen and analyzed by qRT-PCR. Relative gene expression was determined by normalization against Atp5b as described in Materials and Methods and expressed as fold changes relative to untreated control samples (Cntrl). Values are means SEM (n 4 ovaries/cultures). Groups with different letters are statistically different (P .05).

Article Snippet: Follicles were exposed to a range of concentrations of recombinant mouse (rm) GDF9 or recombinant human (rh) BMP15 (R&D Systems Inc., Abingdon, UK).

Techniques: Gene Expression, Cell Culture, Quantitative RT-PCR, Control

Journal: Endocrinology

Article Title: Investigations of TGF-β signaling in preantral follicles of female mice reveal differential roles for bone morphogenetic protein 15.

doi: 10.1210/en.2012-2251

Figure Lengend Snippet: Figure 6. Effect of dorsomorphin (DSM) on preantral follicle growth in the presence of BMP15. Follicles were cultured for 72 h in the presence of BMP15 (250ng/ml) or BMP15 (250ng/ml) DSM (1.0 M) and follicle size was assessed in relation to controls at each time point (a). In a separate experiment follicles were cultured for 48h in the presence of both BMP15 and GDF9 (250ng/ml each) and follicle size was assessed in relation to those cultured with both ligands and DSM (1.0 M) (b). Values are means SEM (n 4 cultures each). *P .05, **P .01 vs Control.

Article Snippet: Follicles were exposed to a range of concentrations of recombinant mouse (rm) GDF9 or recombinant human (rh) BMP15 (R&D Systems Inc., Abingdon, UK).

Techniques: Cell Culture, Control

Journal: Endocrinology

Article Title: Investigations of TGF-β signaling in preantral follicles of female mice reveal differential roles for bone morphogenetic protein 15.

doi: 10.1210/en.2012-2251

Figure Lengend Snippet: Figure 7. Effect of SB505124 on preantral follicle growth in vitro. Follicles were mechanically isolated and cultured individually in 96-well plates for 72 h to test the effects of SB505124 on follicle growth in the presence or absence of GDF9 (a), BMP15 (b) or the combination of BMP15 GDF9 (c). All values shown are means SEM (n 6, 4 and 6 cultures for a, b and c respectively). Comparisons between treatment groups were made within each time point by ANOVA and Bonferroni multiple comparisons test. Groups with different letters are statistically different (P .05).

Article Snippet: Follicles were exposed to a range of concentrations of recombinant mouse (rm) GDF9 or recombinant human (rh) BMP15 (R&D Systems Inc., Abingdon, UK).

Techniques: In Vitro, Isolation, Cell Culture

Journal: Biology of reproduction

Article Title: RNA interference evidence that growth differentiation factor-9 mediates oocyte regulation of cumulus expansion in mice.

doi: 10.1095/biolreprod.104.033357

Figure Lengend Snippet: FIG. 1. Effect of Gdf9 and Bmp15 dsRNA injection on the relative abun- dance of Gdf9 and Bmp15 mRNA expression in fully grown mouse oo- cytes. White bars indicate Gdf9 mRNA levels, and patterned bars indicate Bmp15 mRNA levels. Data are presented with the experimental mean as one; individual bars show the treatment mean 6 SEM. Within a series, bars with different letters are significantly different (P , 0.05).

Article Snippet: After preincubation in 1% fish gelatin in 0.1% Triton-100 PBS, oocytes were incubated with anti-mouse GDF9 polyclonal antibody (R&D Systems Europe).

Techniques: Injection, Expressing

Journal: Biology of reproduction

Article Title: RNA interference evidence that growth differentiation factor-9 mediates oocyte regulation of cumulus expansion in mice.

doi: 10.1095/biolreprod.104.033357

Figure Lengend Snippet: FIG. 2. Effect of Gdf9 dsRNA injection on GDF9 levels in fully grown mouse oocytes. A) Immunoblot assay for GDF9. Molecular weights are indicated in kilodaltons. A strong band of 62.5 kDa was detected in buffer-injected oocytes in comparison with Gdf9 dsRNA-injected oocytes. B) Immunofluo- rescence assay for GDF9. Representative selected images were captured using an Olympus IX70 inverted microscope with a CoolSNAP HQ CCD camera and SoftWorx image capturing software. Shown are a negative control (a; buffer-injected oocyte stained with fluorescein isothiocyanate-con- jugated antibody to goat immunoglobulin G alone), a buffer-injected oocyte (b), and a Gdf9 dsRNA-injected oocyte (c). The inset shows representative lower-magnification images of multiple oocytes. Bar 5 50 mm.

Article Snippet: After preincubation in 1% fish gelatin in 0.1% Triton-100 PBS, oocytes were incubated with anti-mouse GDF9 polyclonal antibody (R&D Systems Europe).

Techniques: Injection, Western Blot, Comparison, Inverted Microscopy, Software, Negative Control, Staining

Journal: Biology of reproduction

Article Title: RNA interference evidence that growth differentiation factor-9 mediates oocyte regulation of cumulus expansion in mice.

doi: 10.1095/biolreprod.104.033357

Figure Lengend Snippet: FIG. 3. Representative photomicrographs showing the effect of GDF9 knockdown on cumulus expansion. a and b) Expanded cumulus shells following 24-h coculture with buffer-injected oocytes (a) and Bmp15 dsRNA-injected oocytes (b) the presence of 0.5 IU/ml of FSH. c) Cumulus shells showing little evidence of expansion following 24-h coculture with Gdf9knockdown oocytes in the presence of 0.5 IU/ml of FSH. Differential interference contrast images were captured using 203 magnification on a Zeiss LSM510 inverted microscope. Bar 5 50 mm.

Article Snippet: After preincubation in 1% fish gelatin in 0.1% Triton-100 PBS, oocytes were incubated with anti-mouse GDF9 polyclonal antibody (R&D Systems Europe).

Techniques: Knockdown, Injection, Inverted Microscopy

Journal: Biology of reproduction

Article Title: RNA interference evidence that growth differentiation factor-9 mediates oocyte regulation of cumulus expansion in mice.

doi: 10.1095/biolreprod.104.033357

Figure Lengend Snippet: FIG. 4. Effect of GDF9 knockdown on Has2 and Ptgs2 mRNA expression in cumulus cells. Cumulus shells were cocultured for 8 h with fully grown oocytes that had been injected with either Gdf9 dsRNA or buffer 24 h earlier. Culture medium was supplemented with 0.5 IU/ml of FSH. White bars indicate Ptgs2 mRNA levels, and patterned bars indicate Has2 mRNA levels. Data are presented with the experimental mean as one; individual bars show the treatment mean 6 SEM. Within a series, bars with different letters are significantly different (P , 0.05).

Article Snippet: After preincubation in 1% fish gelatin in 0.1% Triton-100 PBS, oocytes were incubated with anti-mouse GDF9 polyclonal antibody (R&D Systems Europe).

Techniques: Knockdown, Expressing, Injection

Journal: Endocrinology

Article Title: Heparan sulfate proteoglycans regulate responses to oocyte paracrine signals in ovarian follicle morphogenesis.

doi: 10.1210/en.2012-1181

Figure Lengend Snippet: FIG. 1. Inhibition of cumulus expansion by heparin is restored by GDF9 treatment. Cumulus oocyte complexes were cultured under in vitro maturation conditions (MEM 2% fetal calf serum, 10 mIU/ml FSH, 10 ng/ml Egf) without or with exogenous heparin (300 g/ml) or heparin Gdf9 (500 ng/ml). A–C, Representative images of control (A), heparin-treated (B), and heparin GDF9 (C)-treated COC after 12 h culture. CEI was scored by a trained observer blinded to treatments. Mean SEM of CEI was from three independent experiments (D). Values indicated by different characters are significantly different P 0.001 by one-way ANOVA and Tukey’s post hoc test.

Article Snippet: In the indicated experiments, IVM cultures used either the above control conditions, or included heparin (300 g/ml; Sigma catalog no. H3149) or heparin GDF9 (R&D Systems, Minneapolis, MN; catalog no. 739-G9, 500 ng/ml).

Techniques: Inhibition, Cell Culture, In Vitro, Control

Journal: Endocrinology

Article Title: Heparan sulfate proteoglycans regulate responses to oocyte paracrine signals in ovarian follicle morphogenesis.

doi: 10.1210/en.2012-1181

Figure Lengend Snippet: FIG. 2. Heparin blocks cumulus-specific gene expression and signal transduction mediated by GDF9. A–D, Expression levels of cumulus-specific Has2 (A) and Tnfaip6 (B) or mural granulosa cell specific; Cyp11a1 (C) and Lhcgr (D) transcripts were evaluated after 12 h culture in no treatment (control), heparin (300 g/ml), or heparin GDF9 (500 ng/ml). Expression levels are normalized to rpL19 mRNA loading control assayed in parallel and expressed relative to expression in a pool of whole-ovary calibrator RNA sample. Values indicated by asterisks are significantly different (P 0.05) by using one-way ANOVA and Tukey’s post hoc test. E, Phosphorylated SMAD2 abundance in extracts of COC after 6 h culture in the indicated treatments. Hep, Heparin. Equivalent protein content was loaded in each lane, and -actin was evaluated to confirm equal sample loading. F, Venn diagram illustrating the number of gene transcripts identified in microarray analysis with significantly up- or down-regulated after heparin or SB431542 treatment compared with controls. Numbers indicate the number of transcripts with significantly up- or down-regulated expression level in pairwise comparisons against controls. Overlapping areas represent the number of transcripts whose levels are commonly altered by both treatments. Genes with significantly altered steady-state expression were defined by P 0.05 after ANOVA with step-up adjustment and greater than 2-fold change in expression level (n 4 independent experiments). Mwt, Molecular weight.

Article Snippet: In the indicated experiments, IVM cultures used either the above control conditions, or included heparin (300 g/ml; Sigma catalog no. H3149) or heparin GDF9 (R&D Systems, Minneapolis, MN; catalog no. 739-G9, 500 ng/ml).

Techniques: Gene Expression, Transduction, Expressing, Control, Microarray, Molecular Weight

Journal: Endocrinology

Article Title: Heparan sulfate proteoglycans regulate responses to oocyte paracrine signals in ovarian follicle morphogenesis.

doi: 10.1210/en.2012-1181

Figure Lengend Snippet: FIG. 3. Mouse and human GDF9 bind to heparin with high affinity. IMAC-purified murine (upper panel) or human (lower panel) recombinant GDF9 condition media were incubated with heparin sepharose then eluted in stepwise increasing NaCl concentrations as described in Materials and Methods. Samples were separated on reducing SDS-PAGE gel and GDF9 was detected by Western blot (described in Supplemental Methods) probing with mAb-53 directed against the mature peptide. M, Molecular weight marker (sizes in kilodaltons indicated on the left). SM, Starting material; FT, flowthrough. Arrowheads represent GDF9 (bands at 60 kDa are the intact promature protein; bands at 17 kDa are the processed mature monomer). The lower molecular band is likely a degradation product.

Article Snippet: In the indicated experiments, IVM cultures used either the above control conditions, or included heparin (300 g/ml; Sigma catalog no. H3149) or heparin GDF9 (R&D Systems, Minneapolis, MN; catalog no. 739-G9, 500 ng/ml).

Techniques: Purification, Recombinant, Incubation, SDS Page, Western Blot, Molecular Weight, Marker

Journal: Endocrinology

Article Title: Heparan sulfate proteoglycans regulate responses to oocyte paracrine signals in ovarian follicle morphogenesis.

doi: 10.1210/en.2012-1181

Figure Lengend Snippet: FIG. 6. GDF9 regulates proteoglycan gene expression in COC. Expression of proteoglycans, Glypican 1, Syndecan 4, and Betaglycan, was evaluated in COC after 12 h IVM culture [MEM EGF (10 ng/ml) and FSH (10 mIU/ml)] either with no treatment (control) or treated with heparin (300 g/ml) or heparin GDF9 (500 ng/ml). Expression levels are normalized to rpL19 mRNA loading control assayed in parallel and presented as expression relative to a calibrator sample of whole-ovary RNA. Values are mean SEM of three independent experiments. Those with different characters are significantly different (P 0.05). #, P 0.0549, one-way ANOVA and Tukey’s post hoc test.

Article Snippet: In the indicated experiments, IVM cultures used either the above control conditions, or included heparin (300 g/ml; Sigma catalog no. H3149) or heparin GDF9 (R&D Systems, Minneapolis, MN; catalog no. 739-G9, 500 ng/ml).

Techniques: Gene Expression, Expressing, Control

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 1. Transzonal projections (TZP) density of COCs during IVM with BMP15 (A) and/or GDF9 (B). Phalloidin-FITC fluorescence intensity (mean ± S.E.M.) in the zona pellucida area of COCs after 0h, 6h, 12h and 24h of IVM. A total of 20–30 COCs per group were assessed in 3 replicates. (A) Values with different letters differ significantly (p < 0.05). (B) Different letters between groups in the same timepoint (a,b) and different symbols between timepoints in the same group (α,β,γ for control and GDF9 groups; and σ,τ for GDF9+BMP15 group) indicate significant differences (p < 0.05). (C) Representative confocal images of phalloidin-FITC staining. TZP are observed as thin filaments going through the zona pellucida from the cumulus cells to the oocyte. Scale bar: 100 μm.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques: Fluorescence, Control, Staining

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 2. Mitochondrial activity of MII-oocytes after IVM with BMP15 and/or GDF9. (A) Relative fluorescence intensity (mean ± S.E.M.) of Mitotracker stained oocytes. Columns with different letters differ significantly (p < 0.05). A total of 20–27 oocytes per group were assessed in 3 replicates. (B) Representative images. Scale bar: 100 μm.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques: Activity Assay, Fluorescence, Staining

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 3. ROS and GSH levels after IVM with BMP15 and/or GDF9. Relative fluorescence intensity (mean ± S.E.M.) of (A) ROS and (B) GSH. A total of 24–31 oocytes per group were assessed in 3 replicates. (C) Representative images. Scale bar: 200 μm.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques: Fluorescence

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 4. EGFR expression of MII-oocytes after IVM with BMP15 (A) and/or GDF9 (B). Relative fluorescence intensity (mean ± S.E.M.) of EGFR immunostained oocytes. Columns with different letters differ significantly (p < 0.05). A total of 26–40 oocytes per group were assessed in 4 replicates. (C) Representative images. Scale bar: 30 μm.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques: Expressing, Fluorescence

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 5. EGFR expression of cumulus cells after IVM with BMP15 (A) and/or GDF9 (B). Relative EGFR expression (mean ± S.E.M.). Columns with different letters differ significantly (p < 0.05). Four independent blots were used for relative quantification. (C) Western blot analyses of EGFR expression. EGFR from 70 to 80 COCs/ replicate was quantified by western blotting and standardized to vinculin protein levels.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques: Expressing, Quantitative Proteomics, Western Blot

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 6. Cumulus expansion index after IVM with BMP15 (A) and/or GDF9 (B). Cumulus expansion of each COC was scored on a 0 to 4 scale and cumulus expansion index (mean ± S.E.M) was calculated as the arithmetic mean value scored of all COCs evaluated per replica. Columns with different letters differ significantly (p < 0.05). A total of 367–441(A) and 278–323 (B) oocytes per group were assessed in 5 replicates.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques:

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 7. Effect of IVM supplementation with BMP15 (A) and/or GDF9 (B) on 8-day blastocyst quality after parthenogenic activation (PA). Cell number (mean ± S.E. M.) of the inner cell mass (ICM) and the trophectoderm (TE) of 8-days blastocysts. Columns with different letters differ significantly (p < 0.05). A total of 15–29 blastocysts per group were assessed.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques: Activation Assay

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 8. Effect of IVM supplementation with BMP15 (A) and/or GDF9 (B) on 8-day blastocyst quality after IVF. Cell number (mean ± S.E.M.) of 8-day blastocysts in the inner cell mass (ICM) and the trophectoderm (TE). Columns with different letters differ significantly (p < 0.05). A total of 14–22 blastocysts per group were assessed.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques: