Journal: Nature Communications

Article Title: Transgenerational inheritance of diminished ovarian reserve triggered by prenatal propylparaben exposure in mice

doi: 10.1038/s41467-025-63440-z

Figure Lengend Snippet: a Representative ovarian images for TUNEL staining of F1–F3 offspring mice in Con and PrP groups at 12 w. b The number of TUNEL-positive cells per 1 mm 2 in ovarian sections of F1–F3 offspring mice in Con and PrP groups ( n = 5 biological replicates). Data are presented as mean ± SEM (unpaired two-tailed t -test). c The number of abnormal mitochondria in mice ovaries of F1–F3 generation ( n = 3 biological replicates, two non-overlapping microscope fields per ovary). Data are presented as mean ± SEM (unpaired two-tailed t -test). d Representative TEM images of mice ovaries in F3 generation. Yellow arrow: synapses of granulosa cells. M: mitochondria. The experiment was repeated three times. e Schematic illustration of the ovulation induction. f Representative oocyte images after ovulation induction. g The number of oocytes of F1 and F3 offspring mice in Con and PrP groups after ovulation induction (F1 n = 9; F3 Con n = 6, PrP n = 5 biological replicates). Data are presented as mean ± SEM (unpaired two-tailed t -test). h Representative IHC images showing immunoreactivity against BMP15 in F1, F2 and F3 offspring ovaries. i IHC scores based on IOD/Area of anti-BMP15 in F1, F2 and F3 offspring ovaries ( n = 5 biological replicates). Data are presented as mean ± SEM (unpaired two-tailed t -test). Source data are provided as a Source Data file. Figure 2e is created in BioRender. Wang, S. (2025) https://BioRender.com/bz8sq44 .

Article Snippet: The Antibodies were purchased from different companies: Cell Signaling Technology for ERK (Extracellular Signal-Regulated Kinase; 4695T), p-ERK (Phosphorylated ERK; 4370T), p38 (p38 Mitogen-Activated Protein Kinase; 8690T), p-p38 (Phosphorylated p38; 4511T), JNK (c-Jun N-terminal Kinase; 9252T), and p-JNK (Phosphorylated JNK; 4668T); Abclonal for β-ACTIN (AC038), active-Caspase3 (A19654), BCL-2 (A19693), BAX (A12009), SOD2 (A19576), BMP15 (A7321), CAT (A11777), HSD17B7 (A17157), and CTSK (A1782); Proteintech for FGF18 (60341-1-lg); Immunoway for RhoBTB1 (YT4084); Servicebio for HRP conjugated goat anti-rabbit IgG (GB23303), and Alexa Fluor® 488-conjugated goat anti-rabbit IgG (GB25303).

Techniques: TUNEL Assay, Staining, Two Tailed Test, Microscopy

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: A rare germline BMP15 missense mutation causes hereditary ovarian immature teratoma in human.

doi: 10.1073/pnas.2310409121

Figure Lengend Snippet: Fig. 1. Pathogenic mutant in familial ovarian immature teratoma. (A) Cases from two pedigree of OIT and their corresponding genotypes of BMP15. (B) Pathological diagnosis of corresponding patients. Blue dash-circled areas and arrowheads indicate two to three germ layer-derived tissues in the tumor tissue; neural components and neuroglia in the ectoderm; and the neuroepithelium is connected into pieces in a daisy doughnut or irregular glandular shape. (C) Sanger sequencing results of corresponding patients. (D) Conservation of Arg 88 across various species. (E) Hydrophilicity of Arg 88 across various species. (F) Predicted structural change of c.C262T (MT) mutated BMP15.

Article Snippet: Mouse Model. C57BL/6 mice harboring BMP15 R86C mutation (equivalent to BMP15 R88C in humans) were generated by Cas9/CRISPR- mediated genome editing (Cyagen Biosciences).

Techniques: Mutagenesis, Biomarker Discovery, Derivative Assay, Sequencing

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: A rare germline BMP15 missense mutation causes hereditary ovarian immature teratoma in human.

doi: 10.1073/pnas.2310409121

Figure Lengend Snippet: Fig. 3. C262T mutant significantly stimulates parthenogenetic activation of oocytes. Oocytes were collected from genetically engineered mice harboring the orthologous of BMP15 wild type or C262T variants. Superovulation was induced at 4 wk of age mice. Ten ovaries were assessed from each genotype. The parthenogenetic activation in oocyte was assessed by morphological changes (panel A–C) or histological stain (panel D and E). (A) Two pronucleus were seen in oocyte with spontaneous parthenogenetic activation (yellow arrowhead); (B) two pronucleus and two polar bodies were seen in oocyte with spontaneous parthenogenetic activation (yellow and blue arrowheads, respectively); (C) Two-cell (green arrowheads) was seen with spontaneous parthenogenetic activation; (D) The oocytes appear to have two pronucleus in the HE section (yellow arrowhead); (E) Two polar bodies and one pronucleus appear in the oocyte in the HE section (yellow and blue arrowheads, respectively); (F) Two-cell (green arrowheads) appeared in HE sections; (G) The percentage of parthenogenetically activated oocytes in different BMP15 variants was counted under the Leica DMI3000B inverted biological microscope.

Article Snippet: Mouse Model. C57BL/6 mice harboring BMP15 R86C mutation (equivalent to BMP15 R88C in humans) were generated by Cas9/CRISPR- mediated genome editing (Cyagen Biosciences).

Techniques: Mutagenesis, Activation Assay, Staining, Microscopy

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: A rare germline BMP15 missense mutation causes hereditary ovarian immature teratoma in human.

doi: 10.1073/pnas.2310409121

Figure Lengend Snippet: Fig. 2. C262T mutant affects the protein expression of BMP15. (A) Wild type and C262T mutated BMP15 were overexpressed in HEK293 cells. Amounts of secreted BMP15 (s-BMP15) were compared by western blot. Signal intensity was semi-quantified using ImageJ with the arbitrary unit (P < 0.05). (B) The general BMP15 protein level in a healthy ovarian, BMP15 was highly expressed in both granulosa cells (red arrow) and oocytes (blue arrow) of healthy ovaries; (C) Wild-type BMP15 in sporadic ovarian immature teratoma, BMP15 is highly expressed in the follicular epithelium of tumors (black arrow). (D) C262T variant in familial ovarian immature teratoma. The expression level of the mature BMP15 protein was significantly decreased in immature teratomas carrying the C262T mutation (blue arrow).

Article Snippet: Mouse Model. C57BL/6 mice harboring BMP15 R86C mutation (equivalent to BMP15 R88C in humans) were generated by Cas9/CRISPR- mediated genome editing (Cyagen Biosciences).

Techniques: Mutagenesis, Expressing, Western Blot, Variant Assay

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: A rare germline BMP15 missense mutation causes hereditary ovarian immature teratoma in human.

doi: 10.1073/pnas.2310409121

Figure Lengend Snippet: Fig. 4. H-Ras activated MAPK stimulate spontaneous activation of the oocytes and overgrown of the ovary. (A) Venn diagram showing the enriching strategy of oocytes specific RNA sequencing results. (B) Heat map showing the distinct diverse pattern of differential expressed genes in oocytes with wild-type or mutated BMP15. (C) H-Ras and MEK1 RNA levels in CC, CT, or TT type oocytes. (D) IHC stain showing the BMP15, H-Ras, MEK1, and p-MEK1 protein levels in CC, CT, or TT type ovarian tissues at week 4. (E) (i) Hematoxylin and eosin stain of ovarian tissues at the longest latitude section; blue arrowheads show follicles; (ii) the actual ovary size of CC and CT type at month 6; (iii) average counts of ovarian follicles per slicing section at month 6. PF, primary follicle; SF, secondary follicle; MF, mature follicle; AF, atretic follicle; *** indicates P-value < 0.001. (F) An ovarian teratoma in the CT type ovary. Blue arrowhead shows a digestive tract structure composed of layers of epithelial cells. (G) IHC stain showing the BMP15, H-Ras, MEK1, and p-MEK1 protein levels in a CC type healthy ovary and the CT type ovary with the teratoma at month 6.

Article Snippet: Mouse Model. C57BL/6 mice harboring BMP15 R86C mutation (equivalent to BMP15 R88C in humans) were generated by Cas9/CRISPR- mediated genome editing (Cyagen Biosciences).

Techniques: Activation Assay, RNA Sequencing, Staining, H&E Stain

Journal: Endocrinology

Article Title: Serum Concentrations of Oocyte-Secreted Factors BMP15 and GDF9 During IVF and in Women With Reproductive Pathologies.

doi: 10.1210/en.2019-00264

Figure Lengend Snippet: Figure 1. BMP15 and GDF9 serum ELISA development and validation. (a–d) Standards in the (a and c) BMP15 and (b and d) GDF9 ELISAs were recombinant pro-BMP15 and recombinant mature human GDF9 (GDF9), respectively. (a and b) Specificity was assessed using increasing doses of recombinant human proteins of the TGF-b superfamily (GDF5, GDF6, and GDF7). (c and d) Parallelism between the standards and serum samples was assessed in dose-response curves with pooled serum samples from women of reproductive age (serum QC-1 to 3, top dose 100 mL of serum) and (d) for GDF9, with pro-GDF9. Data are mean 6 SD of (a, b, and d) triplicates and (c) duplicates. OD, optical density.

Article Snippet: Recombinant proteins used as standard preparations in the BMP15andGDF9ELISAswere promature human (h) BMP15 (proBMP15; which is a noncovalent complex–formed postproteolytic cleavage of the pro-domain and mature-domain of BMP15) and mature hGDF9 (R&D Systems, Minneapolis, MN), respectively.

Techniques: Enzyme-linked Immunosorbent Assay, Biomarker Discovery, Recombinant

Journal: Endocrinology

Article Title: Serum Concentrations of Oocyte-Secreted Factors BMP15 and GDF9 During IVF and in Women With Reproductive Pathologies.

doi: 10.1210/en.2019-00264

Figure Lengend Snippet: Figure 2. BMP15 and GDF9 antibody validation. (a–d) Effect of neutralizing the detection and coating antibodies with specific (28A, 53-1, and 72B) and nonspecific (OVA) peptides on the pro-BMP15 and GDF9 standards and/or serum BMP15 QC (QC-4) in the (a and b) BMP15 and (c and d) GDF9 ELISAs. Absorbance was normalized to the standard/sample in the absence of neutralizing peptide and presented as a proportion (%) of the signal. Data are mean 6 SD of (a and c) triplicates and (b and d) duplicates.

Article Snippet: Recombinant proteins used as standard preparations in the BMP15andGDF9ELISAswere promature human (h) BMP15 (proBMP15; which is a noncovalent complex–formed postproteolytic cleavage of the pro-domain and mature-domain of BMP15) and mature hGDF9 (R&D Systems, Minneapolis, MN), respectively.

Techniques: Biomarker Discovery

Journal: Endocrinology

Article Title: Serum Concentrations of Oocyte-Secreted Factors BMP15 and GDF9 During IVF and in Women With Reproductive Pathologies.

doi: 10.1210/en.2019-00264

Figure Lengend Snippet: Figure 3. Serum BMP15 and GDF9 in women at early, middle, and late reproductive age. (a, c, and e) BMP15 and (b, d, and f) GDF9 in 189 women, including asymptomatic women of reproductive age (n 5 6, 22 to 41 y of age), women being treated for infertility (n 5 154, 25 to 52 y of age), perimenopausal women (n 5 3, 48 to 52 y of age), and postmenopausal women (n 5 25, 48 to 66 y of age). Data are presented (a and b) as scatterplots with Spearman correlation coefficients, (c and d) stratified into 5-y age intervals (or 10-y intervals where n , 10 for that interval), or (e and f) as the proportion of women with serum BMP15 or GDF9 above the limit of quantification. (a–d) Dots represent individual women. (a and b) C, premenopausal women; s, postmenopausal women. (c and d) Black dots with bars represent the estimated geometric mean 6 95% CI, dashed horizontal lines indicate assay limit of quantification, and numbers above the x-axis show number of women. **P , 0.01.

Article Snippet: Recombinant proteins used as standard preparations in the BMP15andGDF9ELISAswere promature human (h) BMP15 (proBMP15; which is a noncovalent complex–formed postproteolytic cleavage of the pro-domain and mature-domain of BMP15) and mature hGDF9 (R&D Systems, Minneapolis, MN), respectively.

Techniques:

Journal: Endocrinology

Article Title: Serum Concentrations of Oocyte-Secreted Factors BMP15 and GDF9 During IVF and in Women With Reproductive Pathologies.

doi: 10.1210/en.2019-00264

Figure Lengend Snippet: Figure 4. Follicular phase concentrations of serum BMP15 and GDF9 in women undergoing IVF. (a and c) Individual serum samples throughout GnRH antagonist FSH-stimulated cycles (n 5 15, plus an additional two women for BMP15). Black bars represent blood samples taken prior to exogenous FSH stimulation, and subsequent gray bars are samples taken every 2 to 3 d throughout FSH stimulation. Dashed horizontal lines indicate the assay limit of quantification, and samples with BMP15/GDF9 below this value are denoted “Undetectable.” (b) For women who had a blood sample prior to FSH stimulation (n 5 11), BMP15 concentrations of subsequent samples were normalized to the BMP15 of this baseline sample (day 2 to 3), shown as mean 6 SEM.

Article Snippet: Recombinant proteins used as standard preparations in the BMP15andGDF9ELISAswere promature human (h) BMP15 (proBMP15; which is a noncovalent complex–formed postproteolytic cleavage of the pro-domain and mature-domain of BMP15) and mature hGDF9 (R&D Systems, Minneapolis, MN), respectively.

Techniques:

Journal: Endocrinology

Article Title: Serum Concentrations of Oocyte-Secreted Factors BMP15 and GDF9 During IVF and in Women With Reproductive Pathologies.

doi: 10.1210/en.2019-00264

Figure Lengend Snippet: Figure 5. Serum concentrations of BMP15, GDF9, and AMH and age of women relative to ovarian stimulation type. (a–f) Stimulated cycles were compared: GnRH antagonist and agonist (LDR and short Flare) ovarian stimulation, relative to (a and c) BMP15, (b and d) GDF9, (e) AMH, and (f) age. Data are represented (a, b, e, and f) as estimated geometric mean 6 95% CI (black dots with bars) with gray dots representing individual women, or (c and d) as the proportion of women with serum BMP15 or GDF9 above the assay limit of quantification. (a, b, e, and f) Dashed horizontal lines indicate the assay limit of quantification. Numbers above the x-axes show the number of women. *P , 0.05; ***P , 0.001.

Article Snippet: Recombinant proteins used as standard preparations in the BMP15andGDF9ELISAswere promature human (h) BMP15 (proBMP15; which is a noncovalent complex–formed postproteolytic cleavage of the pro-domain and mature-domain of BMP15) and mature hGDF9 (R&D Systems, Minneapolis, MN), respectively.

Techniques:

Journal: Endocrinology

Article Title: Serum Concentrations of Oocyte-Secreted Factors BMP15 and GDF9 During IVF and in Women With Reproductive Pathologies.

doi: 10.1210/en.2019-00264

Figure Lengend Snippet: Figure 6. Serum concentrations of BMP15, GDF9, and AMH and age of stimulated and unstimulated women during infertility treatment. All stimulated cycles were combined and compared with unstimulated cycles, including natural cycle monitoring or IUI without FSH, hormone replacement therapy, or ovulation induction. Data are represented (a, b, e, and f) as estimated geometric mean 6 95% CI (black dots with bars), with gray dots representing individual women, or (c and d) as the proportion of women with serum BMP15 or GDF9 above the assay limit of quantification. Dashed horizontal lines indicate the assay limit of quantification (a, b, e, and f). Numbers above the x-axes show the number of women.

Article Snippet: Recombinant proteins used as standard preparations in the BMP15andGDF9ELISAswere promature human (h) BMP15 (proBMP15; which is a noncovalent complex–formed postproteolytic cleavage of the pro-domain and mature-domain of BMP15) and mature hGDF9 (R&D Systems, Minneapolis, MN), respectively.

Techniques:

Journal: Endocrinology

Article Title: Serum Concentrations of Oocyte-Secreted Factors BMP15 and GDF9 During IVF and in Women With Reproductive Pathologies.

doi: 10.1210/en.2019-00264

Figure Lengend Snippet: Figure 7. Serum BMP15, GDF9 and AMH relative to the number of oocytes retrieved following ovarian stimulation. (a–f) Women with cancelled cycles due to too few follicles developed with stimulation and with increasing numbers of oocytes retrieved (binned 1 to 5, 6 to 10, 11 to 15, .15 oocytes) were compared with respect to BMP15, GDF9, and AMH. Data are represented as (a–c and g–i) estimated geometric mean 6 95% CI (black dots with bars), with gray dots representing individual women, or (d–f) as the proportion of women with serum BMP15 or GDF9 above the assay limit of quantification. Dashed horizontal lines indicate the assay limit of quantification (a–c and g–i). Numbers above the x-axes show number of women. ***P , 0.001.

Article Snippet: Recombinant proteins used as standard preparations in the BMP15andGDF9ELISAswere promature human (h) BMP15 (proBMP15; which is a noncovalent complex–formed postproteolytic cleavage of the pro-domain and mature-domain of BMP15) and mature hGDF9 (R&D Systems, Minneapolis, MN), respectively.

Techniques:

Journal: Endocrinology

Article Title: Serum Concentrations of Oocyte-Secreted Factors BMP15 and GDF9 During IVF and in Women With Reproductive Pathologies.

doi: 10.1210/en.2019-00264

Figure Lengend Snippet: Figure 8. Relationship between serum biomarker concentration and oocyte number in women with and without polycystic ovaries and PCOS. (a–c) Regression modeling of (a) BMP15, (b) GDF9, and (c) AMH concentrations relative to the number of oocytes retrieved following ovarian stimulation in women without PCOS (n; solid regression lines) and women with PCOS (4; dashed regression lines). Tables below each figure indicate the relative change (Rel. change) in biomarker concentration associated with increasing number of oocytes in each patient group and associated P values. The P(int) term describes the significance of the interaction between

Article Snippet: Recombinant proteins used as standard preparations in the BMP15andGDF9ELISAswere promature human (h) BMP15 (proBMP15; which is a noncovalent complex–formed postproteolytic cleavage of the pro-domain and mature-domain of BMP15) and mature hGDF9 (R&D Systems, Minneapolis, MN), respectively.

Techniques: Biomarker Discovery, Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Long-Term Changes in Ovarian Follicles of Gilts Exposed Neonatally to Methoxychlor: Effects on Oocyte-Derived Factors, Anti-Müllerian Hormone, Follicle-Stimulating Hormone, and Cognate Receptors

doi: 10.3390/ijms23052780

Figure Lengend Snippet: The mRNA expression and protein abundance of GDF9 ( a , a’ ), BMP15 ( b , b’ ) in preantral follicles, as well as BMPR2 ( c , c’ ) in preantral and small antral follicles obtained from the control (CTR) and methoxychlor (MXC-) treated gilts. The mRNA expression (quantitative real-time PCR) is presented relative to GAPDH as mean ± SEM ( a – c ). Relative protein abundance was measured by the densitometric method and expressed as the ratio relative to β-actin abundance (mean ± SEM; a’ – c’ ). The fragment of membranes with bands corresponding to predicted molecular weights is shown in the above graphs. Asterisks on the graphs denote significant differences between the CTR and MXC-treated animals (for preantral follicles pool n = 5, for small antral follicle n = 15, * p < 0.05, ** p < 0.01, *** p < 0.001, Mann–Whitney U test). ( d ) Localization of GDF9 and BMP15 in preantral (primordial, primary, and secondary follicle) as well as localization of BMPR2 in preantral and small antral follicles of the CTR and MXC-treated pigs. GDF9-, BMP15-, and BMPR2-positive staining was observed in oocytes (asterisks) and granulosa cells (arrows) in primordial, primary, and secondary follicles, while in small antral follicles, BMPR2-positive staining was detected in granulosa cells (arrows) and theca cells (arrowheads) in both examined groups. All sections were counterstained with hematoxylin QS. No positive staining was observed in the negative control sections ( d , insets). Prim—primordial follicles; PF—primary follicles; SF—secondary follicles; AF—antral follicles. Bars = 50 µm.

Article Snippet: Real-time quantitative PCR (qPCR) of cDNAs was performed using TaqMan Gene Expression Master Mix (Applied Biosystems) and porcine-specific TaqMan Gene Expression Assay (Applied Biosystems) for: AMH (assay ID: Ss03383931_m1), AMHR2 (assay ID: Ss04321772_m1), BMP15 (assay ID: Ss04248749_s1), BMPR1A (assay ID: Ss04248558_m1), BMPR1B (assay ID: Ss03380019_u1), BMPR2 (assay ID: Ss04248598_m1), FSHR (assay ID: Ss03384581_u1), GDF9 (assay ID: Ss03391680_m1), TGFBR1 (assay ID: Ss03392139_m1), and glyceraldehyde-3-phosphate dehydrogenase (( GAPDH ) assay ID: Ss03373286_u1) as endogenous control, following the manufacturers’ instructions.

Techniques: Expressing, Quantitative Proteomics, Control, Real-time Polymerase Chain Reaction, MANN-WHITNEY, Staining, Negative Control

Journal: International Journal of Molecular Sciences

Article Title: Long-Term Changes in Ovarian Follicles of Gilts Exposed Neonatally to Methoxychlor: Effects on Oocyte-Derived Factors, Anti-Müllerian Hormone, Follicle-Stimulating Hormone, and Cognate Receptors

doi: 10.3390/ijms23052780

Figure Lengend Snippet: List of primary antibodies used for Western blot and immunohistochemistry.

Article Snippet: Real-time quantitative PCR (qPCR) of cDNAs was performed using TaqMan Gene Expression Master Mix (Applied Biosystems) and porcine-specific TaqMan Gene Expression Assay (Applied Biosystems) for: AMH (assay ID: Ss03383931_m1), AMHR2 (assay ID: Ss04321772_m1), BMP15 (assay ID: Ss04248749_s1), BMPR1A (assay ID: Ss04248558_m1), BMPR1B (assay ID: Ss03380019_u1), BMPR2 (assay ID: Ss04248598_m1), FSHR (assay ID: Ss03384581_u1), GDF9 (assay ID: Ss03391680_m1), TGFBR1 (assay ID: Ss03392139_m1), and glyceraldehyde-3-phosphate dehydrogenase (( GAPDH ) assay ID: Ss03373286_u1) as endogenous control, following the manufacturers’ instructions.

Techniques: Western Blot, Immunohistochemistry