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: 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: Anti-BMP15 , 1:500 , 1:50 , rabbit polyclonal , Biorbyt Ltd., Cambridge, UK (orb377952, Lot no. CQ2185).

Techniques: Western Blot, Immunohistochemistry

Journal: Journal of Traditional Chinese Medicine

Article Title: Efficacy of Bushenjianpi prescription on autoimmune premature ovarian failure in mice

doi: 10.1016/S0254-6272(17)30321-7

Figure Lengend Snippet: Figure 3 Immunohistochemical analysis of the distinct presence of Cx43 and BMP-15 in the oophorons of each group (× 200) A-F: Cx43; G-L: BMP-15. A, G: control group; B, H: model group; C, I: positive group; D, J: low dose of BSJPP group; E, K: moderate dose of BSJPP group; F, L: high dose of BSJPP group. Positive group treated with premarin (0.03 mg/kg) once daily for 90 d. Con- trol and model group received the same volume of distilled water. Low (8.1 mg/kg), moderate (16.2 mg/kg, the dose used in vivo in human studies), and high (32.4 mg/kg) dose of BSJPP groups treated by gastrogavage once daily for 90 d. Immunohistochemi- cal analysis clearly revealed the presence of Cx43 and BMP-15 in the oophorons of control mice and those treated with BSJPP (D, E) and premarin compared with the model group. BSJPP: Bushenjianpi prescription; Cx43: connexin 43.

Article Snippet: Anti-connexin 43 (Cx43) antibody (product lot No. BA1727) and anti-bone morphogenetic protein 15 (BMP-15) antibody (product lot No. BA2018) were purchased from Boster Biological Engineering (Wuhan, China).

Techniques: Immunohistochemical staining, Control, In Vivo

Journal: Journal of Traditional Chinese Medicine

Article Title: Efficacy of Bushenjianpi prescription on autoimmune premature ovarian failure in mice

doi: 10.1016/S0254-6272(17)30321-7

Figure Lengend Snippet: Figure 4 Expression Cx43 and BMP-15 mRNA in the ovaries of mice in each group. 1: control group; 2: model group; 3: positive group; 4: low dose of BSJPP group; 5: moderate dose of BSJPP group; 6: high dose of BSJPP group. Positive group treated with pre- marin (0.03 mg/kg) once daily for 90 d. Control and model group received the same volume of distilled water. Low (8.1 mg/kg), moderate (16.2 mg/kg, the dose used in vivo in human studies), and high (32.4 mg/kg) dose of BSJPP groups treated by gastrogavage once daily for 90 d. Compared with the model group, significantly upregulated expression of both Cx43 and BMP-15 was detected in the control group and the groups treated with BSJPP (M and L groups). BSJPP: Bushenjianpi prescription; Cx43: connexin 43.

Article Snippet: Anti-connexin 43 (Cx43) antibody (product lot No. BA1727) and anti-bone morphogenetic protein 15 (BMP-15) antibody (product lot No. BA2018) were purchased from Boster Biological Engineering (Wuhan, China).

Techniques: Expressing, Control, In Vivo

Journal: Frontiers in Cell and Developmental Biology

Article Title: Bone Morphogenetic Protein 15 Knockdown Inhibits Porcine Ovarian Follicular Development and Ovulation

doi: 10.3389/fcell.2019.00286

Figure Lengend Snippet: Generation of the BMP15 knockdown pig model. (A) Diagram of shRNA expression vector. Synthesized hU6-BMP15 shRNA fragment was inserted downstream of EGFP expression cassette on pEGFP-N1 vector. (B) RNA interference efficiency of five BMP15 shRNAs was examined by a dual-luciferase reporter system after 48 h transfection of h293T cells. NC, random shRNA plasmid. (C) PCR analysis of the muscle tissue proved that the integrated Bmp15 shRNA fragment had been transmitted to F1 gilts. +, TG gilt; -, sibling WT gilt; M, DNA Maker. (D) Southern blot analysis showed slightly less than 10 copies of constructed plasmids integrated in both F0 and F1 TG pigs, which was consistent with the result of about seven copies of qPCR analysis (data not shown). DNA with pEGFP- Bmp15 shRNA plasmid copies of 10, 20, and 40 were used as the positive control. (E) qPCR analysis of BMP15 mRNA level in 365 days old transgenic ovaries with two different phenotypes (TGF and TGS). TGF, transgenic ovary with many visible antral follicles on ovarian surface. TGS, transgenic ovary with streak phenotype. ∗ p < 0.05. (F) Western blot analysis of BMP15 protein level in postnatal 30-day old TG ovaries. Three prominent, distinct bands were observed corresponding to apparent molecular weights of 34 kDa, 27 kDa, and 15 kDa. (G) Quantitative analysis of BMP15 protein levels based on the band intensity using Image J software. (H) F1 TG gilt showed a visible intense GFP fluorescence on the toes and muscle while subjected to sunlight.

Article Snippet: Antibodies of BMP15 (Eterlife, EL806306-100), GDF9 (Eterlife, EL901942-100), FSHR (Eterlife, EL912710), luteinizing hormone receptor (LHR) (Eterlife, EL904141), Caspase3 (Abcam, ab13847), 3βHSD (Abcam, ab154385), p-Smad1/5 (CST, 9516), Ki67 (Abcam, ab15580) and LC3B (Arigo, ARG55799) were diluted 1:100 with PBS.

Techniques: Knockdown, shRNA, Expressing, Plasmid Preparation, Synthesized, Luciferase, Transfection, Southern Blot, Construct, Positive Control, Transgenic Assay, Western Blot, Software, Fluorescence

Journal: Frontiers in Cell and Developmental Biology

Article Title: Bone Morphogenetic Protein 15 Knockdown Inhibits Porcine Ovarian Follicular Development and Ovulation

doi: 10.3389/fcell.2019.00286

Figure Lengend Snippet: BMP15 knockdown-induced changes in ovarian and follicular development. (A) Representative photograms of ovaries collected from gilts of different ages showing reduced ovarian size and less visible follicles on the surface of TG ovaries as compared to ovaries from WT sibling. Bilateral TG ovaries were significantly different in size at ages of 200 and 365 days. Two ovarian phenotypes were found in TG ovaries, TGF ovaries had many visible large antral follicles on the ovarian surface. TGS ovaries contained none or less than three visible antral follicles (red arrows). (B) Histological observation of TGS ovaries showed that 110- and 200-day old TGS ovary presented major primary-like follicles sparsely scattered on the cortex (green arrows), while 365-day old TGS ovarian section was predominantly occupied by degraded secondary follicles (black arrow). (C) On the 200-day TGF ovarian section, decreased number of follicles and enlarged antral follicles (black arrow) was observed. In addition, the degradation of GCs in abnormally organized GC layer structure of secondary follicles was observed (black arrows). (D) In 30- and 80-day old TGF ovaries, drastically decreased number of early stage follicles led to thinner ovarian cortex (blue line and blue arrows). (E) Comparison of three ovarian phenotypes at the age of 110 days showed less number of early stage follicles in TGF ovarian cortex, and the minimum number of follicles in TGS ovaries. (F) Markedly reduced proportion of normal secondary follicles in the TGF ovaries. Secondary follicles in three sections of each ovary were counted. (G) Representative images of abnormal TGF secondary follicles (black arrows), including multiovular follicle with highly irregularly organized theca cell layers (i); follicle with oocyte-free structure, and abnormally thickened zona pellucida surrounded by highly degraded GCs (ii); follicle with abnormally thickened theca layers (iii); follicle with enlarged oocyte surrounded by highly irregularly organized GC layers with holes formed by degradation of GCs (iv). (H) TGF follicle showed larger oocyte in the early secondary follicle stage (black arrow head). (I) Smaller GCs were loosely organized in TGF antral follicles (black arrow head).

Article Snippet: Antibodies of BMP15 (Eterlife, EL806306-100), GDF9 (Eterlife, EL901942-100), FSHR (Eterlife, EL912710), luteinizing hormone receptor (LHR) (Eterlife, EL904141), Caspase3 (Abcam, ab13847), 3βHSD (Abcam, ab154385), p-Smad1/5 (CST, 9516), Ki67 (Abcam, ab15580) and LC3B (Arigo, ARG55799) were diluted 1:100 with PBS.

Techniques: Knockdown, Comparison

Journal: Frontiers in Cell and Developmental Biology

Article Title: Bone Morphogenetic Protein 15 Knockdown Inhibits Porcine Ovarian Follicular Development and Ovulation

doi: 10.3389/fcell.2019.00286

Figure Lengend Snippet: Abnormal TGF follicles showed premature luteinization and impaired oocyte quality. (A) Immunostaining of ovarian sections indicated that the expression of BMP15 decreased remarkably in TGS abnormal follicles, but was only slightly reduced in TGF follicles, compared with WT follicles. FSHR shared a similar expression pattern with BMP15, and the expression pattern of LHR was different from that of BMP15. It expressed higher in TGF GCs of both preantral and antral follicles. Scale bar = 100 μm. (B) Follicular cell apoptosis, proliferation, and premature luteinization were evaluated by immunostaining with Caspase3, Ki67, and 3βHSD, respectively. Notably, higher expression level of 3βHSD was discovered in abnormal TGF follicles. However, the expression of Caspase3 and Ki67 in abnormal TGF follicles was not significantly different from that in WT follicles. Scale bar = 100 μm. (C) Immunofluorescence images demonstrates intensive expression of autophagy-related protein, LC3B, in oocytes of normal follicles of TGF and WT ovaries, but it was barely expressed in oocytes of abnormal follicles of TG (TGF and TGS) ovaries. Purple arrow, oocytes in normal follicles; Orange arrow, oocytes in abnormal follicles. Scale bar = 100 μm.

Article Snippet: Antibodies of BMP15 (Eterlife, EL806306-100), GDF9 (Eterlife, EL901942-100), FSHR (Eterlife, EL912710), luteinizing hormone receptor (LHR) (Eterlife, EL904141), Caspase3 (Abcam, ab13847), 3βHSD (Abcam, ab154385), p-Smad1/5 (CST, 9516), Ki67 (Abcam, ab15580) and LC3B (Arigo, ARG55799) were diluted 1:100 with PBS.

Techniques: Immunostaining, Expressing, Immunofluorescence

Journal: Frontiers in Cell and Developmental Biology

Article Title: Bone Morphogenetic Protein 15 Knockdown Inhibits Porcine Ovarian Follicular Development and Ovulation

doi: 10.3389/fcell.2019.00286

Figure Lengend Snippet: Interest genes that related to COCs function.

Article Snippet: Antibodies of BMP15 (Eterlife, EL806306-100), GDF9 (Eterlife, EL901942-100), FSHR (Eterlife, EL912710), luteinizing hormone receptor (LHR) (Eterlife, EL904141), Caspase3 (Abcam, ab13847), 3βHSD (Abcam, ab154385), p-Smad1/5 (CST, 9516), Ki67 (Abcam, ab15580) and LC3B (Arigo, ARG55799) were diluted 1:100 with PBS.

Techniques: Membrane

Journal: Frontiers in Cell and Developmental Biology

Article Title: Bone Morphogenetic Protein 15 Knockdown Inhibits Porcine Ovarian Follicular Development and Ovulation

doi: 10.3389/fcell.2019.00286

Figure Lengend Snippet: Summary of biological functions and possible regulatory mechanism of BMP15 in TGF follicular development. Compared to WT gilt, knockdown of BMP15 caused remarkable reduction of follicle number during the TGF follicular developmental process, accompanied by impaired oocyte quality, degradation, and premature luteinization in TGF preantral follicles. This may lead to increased expression of LHR and dramatically decreased E2 production in TGF antral follicles, resulting in lack of dominant follicle selection but abnormally enlarged antral follicles, presenting a dysovulation phenotype. Decreased E2 production in TGF antral follicles may have a negative feedback to pituitary to increase the expression of Fsh . However, the decreased expression of Fsh receptor, Fshr , in GCs beyond the PF stage, by knocking down BMP15, attenuates the stimulation of antral follicles growth. In addition, BMP15 knockdown leads to the inhibition of cell proliferation and differentiation, declined oocyte quality, and meiotic maturation during TGF follicular development, contributing to the appearance of enlarged follicle in TGF ovary and dysovulation. Colored ellipses represent results based on morphological observation and molecular examination. Dotted ellipses represent results based on transcriptomic analysis.

Article Snippet: Antibodies of BMP15 (Eterlife, EL806306-100), GDF9 (Eterlife, EL901942-100), FSHR (Eterlife, EL912710), luteinizing hormone receptor (LHR) (Eterlife, EL904141), Caspase3 (Abcam, ab13847), 3βHSD (Abcam, ab154385), p-Smad1/5 (CST, 9516), Ki67 (Abcam, ab15580) and LC3B (Arigo, ARG55799) were diluted 1:100 with PBS.

Techniques: Knockdown, Expressing, Selection, Inhibition

Journal: Frontiers in Cell and Developmental Biology

Article Title: Granulosa Cells Improved Mare Oocyte Cytoplasmic Maturation by Providing Collagens

doi: 10.3389/fcell.2022.914735

Figure Lengend Snippet: Effects of GC co-culture on cumulus cell expansion and BMP15 expression in mare MII oocytes. Morphology of COCs matured without GCs [CONTROL, (A) ] or with LFGCs [LFGC + O, (B) ], or SFGCs [SFGC + O, (C) ] for 36 h was examined. (D) The degree of cumulus cell expansion was assessed after IVM (20 oocytes per group). Scale bar = 150 μm. (E) Immunofluorescence detection of BMP15 in MII oocytes derived from the CONTROL, LFGC + O, and SFGC + O groups (30 oocytes per group). (F) Average optical intensity was measured using ImageJ. Values are presented as means ± SEM. * p < 0.05, ** p < 0.01.

Article Snippet: Then, the samples were incubated overnight at 4°C with primary antibodies against BMP15 (1:1000, ABclonal Technology Co., Ltd., Woburn, MA, United States) according to the manufacturer’s instructions.

Techniques: Co-Culture Assay, Expressing, Control, Immunofluorescence, Derivative Assay