Journal: International Journal of Molecular Sciences

Article Title: Classification of Atretic Small Antral Follicles in the Human Ovary

doi: 10.3390/ijms242316846

Figure Lengend Snippet: Granulosa and theca cells in healthy small antral follicles and type 1 atretic follicles from cis female donors. ( A ) Immunofluorescence for CYP19A1, AMH, AR, and CDH1 in healthy small antral follicles and type 1 atretic follicles from cis female donors. White dashed line depicts the basement membrane. ( B ) Histological sections (hematoxylin-eosin-stained) of adult ovaries from trans masculine donors depicting a healthy small antral follicle and type 1 atretic follicle containing the oocyte (yellow arrow), magnified in the top-right part. ( C ) Immunofluorescence for STAR and ACTA2 (top panels) and STAR and GSTA1 (bottom panels) in healthy small antral follicles and type 1 atretic follicles from cis female donors. White dashed line depicts the basement membrane between granulosa cells (GC) and internal theca cells (iTC); blue dashed line depicts the border between iTC and external theca cells (eTC). ( D ) Immunofluorescence for CD68 and COLIV in healthy small antral follicles and type 1 atretic follicles from cis female donors. Yellow arrow points to CD68+ macrophages. White dashed line depicts the basement membrane.

Article Snippet: The primary antibodies used were mouse anti-CYP19A1 (1:100, sc-374176, Santa Cruz, Dallas, TX, USA), mouse anti-AR (1:100, sc-7305, Santa Cruz), mouse anti-AMH (1:100, MCA2246T, BioRad, Hercules, CA, USA), rabbit anti-CDH1 (1:100, CST3195S, Cell Signaling, Danvers, MA, USA), rabbit anti-MKI67 (1:100, ab15580, Abcam, Cambridge, UK), rabbit anti-cCASP3 (Asp175) (1:100, CST9661S, Cell Signaling), rabbit anti-COLLIV (1:100, AB748, Merck), mouse anti-CD68 (1:100, M087629-2, DAKO, Glostrup, Denmark), rabbit anti-ACTA2 (1:100, ab5694, Abcam), mouse anti-STAR (D-2) (1:100, sc-166821, Santa Cruz), rabbit anti-GSTA1 (1:100, HPA053817, Atlas Antibodies, Bromma, Sweden).

Techniques: Immunofluorescence, Staining

Journal: International Journal of Molecular Sciences

Article Title: Classification of Atretic Small Antral Follicles in the Human Ovary

doi: 10.3390/ijms242316846

Figure Lengend Snippet: Type 2 atretic follicles in human ovaries. ( A ) Overview of the main follicle-associated cell types, including granulosa cells (GC) and theca cells (TC), and the structure of type 2 (2a and 2b) atretic follicles. In the magnified hematoxylin-eosin-stained areas: yellow arrow points to a TC with hyaline area; green asterisk in antral cavity marks GC showing pyknotic nuclei; and yellow dashed line marks the basement membrane. ( B ) Immunofluorescence for MKI67, CYP19A1, AR, AMH, CDH1, and type 2a atretic follicles from cis female donors. White dashed line depicts the basement membrane. ( C ) Immunofluorescence staining for STAR and ACTA2 (left panels) and STAR and GSTA1 (right panels) in type 2 atretic follicles from cis female donors. In the magnified areas: yellow arrow points to the transition between ACTA2+ and ACTA2-areas bordering the antral cavity; white dashed line depicts the basement membrane; blue dashed line depicts the border between internal theca cells (iTCs) and external theca cells (eTC). ( D ) Immunofluorescence for CD68 and COLIV in healthy small antral follicles and type 2 atretic follicles from cis female donors. Yellow arrows point to CD68+ macrophages. White dashed line depicts the basement membrane.

Article Snippet: The primary antibodies used were mouse anti-CYP19A1 (1:100, sc-374176, Santa Cruz, Dallas, TX, USA), mouse anti-AR (1:100, sc-7305, Santa Cruz), mouse anti-AMH (1:100, MCA2246T, BioRad, Hercules, CA, USA), rabbit anti-CDH1 (1:100, CST3195S, Cell Signaling, Danvers, MA, USA), rabbit anti-MKI67 (1:100, ab15580, Abcam, Cambridge, UK), rabbit anti-cCASP3 (Asp175) (1:100, CST9661S, Cell Signaling), rabbit anti-COLLIV (1:100, AB748, Merck), mouse anti-CD68 (1:100, M087629-2, DAKO, Glostrup, Denmark), rabbit anti-ACTA2 (1:100, ab5694, Abcam), mouse anti-STAR (D-2) (1:100, sc-166821, Santa Cruz), rabbit anti-GSTA1 (1:100, HPA053817, Atlas Antibodies, Bromma, Sweden).

Techniques: Staining, Immunofluorescence

Journal: International Journal of Molecular Sciences

Article Title: Classification of Atretic Small Antral Follicles in the Human Ovary

doi: 10.3390/ijms242316846

Figure Lengend Snippet: Type 3 atretic follicles in human ovaries. ( A ) Overview of the main follicle-associated cell types, such as granulosa cells (GC) and theca cells (TC), and follicular structure in type 3 (3a and 3b) atretic follicles from adult ovaries. In the magnified hematoxylin-eosin-stained areas: yellow dashed line marks the place of the basement membrane; yellow asterisks mark the place where the basement membrane was ruptured; and black arrow points to the remnant of the antral cavity. ( B ) Immunofluorescence for CD68, COLIV, ACTA2, STAR, and GSTA1 in type 3 atretic follicles from cis female donors. White dashed line marks the basement membrane. Yellow asterisks mark the place where the basement membrane was ruptured. ( C ) Histological sections (haematoxylin-eosin-stained) of adult ovaries depicting type 2 and type 3 atretic follicles containing the oocyte or oocyte remnants (yellow arrow), magnified in the top part.

Article Snippet: The primary antibodies used were mouse anti-CYP19A1 (1:100, sc-374176, Santa Cruz, Dallas, TX, USA), mouse anti-AR (1:100, sc-7305, Santa Cruz), mouse anti-AMH (1:100, MCA2246T, BioRad, Hercules, CA, USA), rabbit anti-CDH1 (1:100, CST3195S, Cell Signaling, Danvers, MA, USA), rabbit anti-MKI67 (1:100, ab15580, Abcam, Cambridge, UK), rabbit anti-cCASP3 (Asp175) (1:100, CST9661S, Cell Signaling), rabbit anti-COLLIV (1:100, AB748, Merck), mouse anti-CD68 (1:100, M087629-2, DAKO, Glostrup, Denmark), rabbit anti-ACTA2 (1:100, ab5694, Abcam), mouse anti-STAR (D-2) (1:100, sc-166821, Santa Cruz), rabbit anti-GSTA1 (1:100, HPA053817, Atlas Antibodies, Bromma, Sweden).

Techniques: Staining, Immunofluorescence

Journal: International Journal of Molecular Sciences

Article Title: Classification of Atretic Small Antral Follicles in the Human Ovary

doi: 10.3390/ijms242316846

Figure Lengend Snippet: Expression levels of markers of interest in specific cell types in atretic small antral follicles.

Article Snippet: The primary antibodies used were mouse anti-CYP19A1 (1:100, sc-374176, Santa Cruz, Dallas, TX, USA), mouse anti-AR (1:100, sc-7305, Santa Cruz), mouse anti-AMH (1:100, MCA2246T, BioRad, Hercules, CA, USA), rabbit anti-CDH1 (1:100, CST3195S, Cell Signaling, Danvers, MA, USA), rabbit anti-MKI67 (1:100, ab15580, Abcam, Cambridge, UK), rabbit anti-cCASP3 (Asp175) (1:100, CST9661S, Cell Signaling), rabbit anti-COLLIV (1:100, AB748, Merck), mouse anti-CD68 (1:100, M087629-2, DAKO, Glostrup, Denmark), rabbit anti-ACTA2 (1:100, ab5694, Abcam), mouse anti-STAR (D-2) (1:100, sc-166821, Santa Cruz), rabbit anti-GSTA1 (1:100, HPA053817, Atlas Antibodies, Bromma, Sweden).

Techniques: Expressing, TUNEL Assay

Journal: International Journal of Molecular Sciences

Article Title: Classification of Atretic Small Antral Follicles in the Human Ovary

doi: 10.3390/ijms242316846

Figure Lengend Snippet: Granulosa and theca cells in healthy small antral follicles and type 1 atretic follicles from cis female donors. ( A ) Immunofluorescence for CYP19A1, AMH, AR, and CDH1 in healthy small antral follicles and type 1 atretic follicles from cis female donors. White dashed line depicts the basement membrane. ( B ) Histological sections (hematoxylin-eosin-stained) of adult ovaries from trans masculine donors depicting a healthy small antral follicle and type 1 atretic follicle containing the oocyte (yellow arrow), magnified in the top-right part. ( C ) Immunofluorescence for STAR and ACTA2 (top panels) and STAR and GSTA1 (bottom panels) in healthy small antral follicles and type 1 atretic follicles from cis female donors. White dashed line depicts the basement membrane between granulosa cells (GC) and internal theca cells (iTC); blue dashed line depicts the border between iTC and external theca cells (eTC). ( D ) Immunofluorescence for CD68 and COLIV in healthy small antral follicles and type 1 atretic follicles from cis female donors. Yellow arrow points to CD68+ macrophages. White dashed line depicts the basement membrane.

Article Snippet: The primary antibodies used were mouse anti-CYP19A1 (1:100, sc-374176, Santa Cruz, Dallas, TX, USA), mouse anti-AR (1:100, sc-7305, Santa Cruz), mouse anti-AMH (1:100, MCA2246T, BioRad, Hercules, CA, USA), rabbit anti-CDH1 (1:100, CST3195S, Cell Signaling, Danvers, MA, USA), rabbit anti-MKI67 (1:100, ab15580, Abcam, Cambridge, UK), rabbit anti-cCASP3 (Asp175) (1:100, CST9661S, Cell Signaling), rabbit anti-COLLIV (1:100, AB748, Merck), mouse anti-CD68 (1:100, M087629-2, DAKO, Glostrup, Denmark), rabbit anti-ACTA2 (1:100, ab5694, Abcam), mouse anti-STAR (D-2) (1:100, sc-166821, Santa Cruz), rabbit anti-GSTA1 (1:100, HPA053817, Atlas Antibodies, Bromma, Sweden).

Techniques: Immunofluorescence, Staining

Journal: International Journal of Molecular Sciences

Article Title: Classification of Atretic Small Antral Follicles in the Human Ovary

doi: 10.3390/ijms242316846

Figure Lengend Snippet: Type 2 atretic follicles in human ovaries. ( A ) Overview of the main follicle-associated cell types, including granulosa cells (GC) and theca cells (TC), and the structure of type 2 (2a and 2b) atretic follicles. In the magnified hematoxylin-eosin-stained areas: yellow arrow points to a TC with hyaline area; green asterisk in antral cavity marks GC showing pyknotic nuclei; and yellow dashed line marks the basement membrane. ( B ) Immunofluorescence for MKI67, CYP19A1, AR, AMH, CDH1, and type 2a atretic follicles from cis female donors. White dashed line depicts the basement membrane. ( C ) Immunofluorescence staining for STAR and ACTA2 (left panels) and STAR and GSTA1 (right panels) in type 2 atretic follicles from cis female donors. In the magnified areas: yellow arrow points to the transition between ACTA2+ and ACTA2-areas bordering the antral cavity; white dashed line depicts the basement membrane; blue dashed line depicts the border between internal theca cells (iTCs) and external theca cells (eTC). ( D ) Immunofluorescence for CD68 and COLIV in healthy small antral follicles and type 2 atretic follicles from cis female donors. Yellow arrows point to CD68+ macrophages. White dashed line depicts the basement membrane.

Article Snippet: The primary antibodies used were mouse anti-CYP19A1 (1:100, sc-374176, Santa Cruz, Dallas, TX, USA), mouse anti-AR (1:100, sc-7305, Santa Cruz), mouse anti-AMH (1:100, MCA2246T, BioRad, Hercules, CA, USA), rabbit anti-CDH1 (1:100, CST3195S, Cell Signaling, Danvers, MA, USA), rabbit anti-MKI67 (1:100, ab15580, Abcam, Cambridge, UK), rabbit anti-cCASP3 (Asp175) (1:100, CST9661S, Cell Signaling), rabbit anti-COLLIV (1:100, AB748, Merck), mouse anti-CD68 (1:100, M087629-2, DAKO, Glostrup, Denmark), rabbit anti-ACTA2 (1:100, ab5694, Abcam), mouse anti-STAR (D-2) (1:100, sc-166821, Santa Cruz), rabbit anti-GSTA1 (1:100, HPA053817, Atlas Antibodies, Bromma, Sweden).

Techniques: Staining, Immunofluorescence

Journal: International Journal of Molecular Sciences

Article Title: Classification of Atretic Small Antral Follicles in the Human Ovary

doi: 10.3390/ijms242316846

Figure Lengend Snippet: Expression levels of markers of interest in specific cell types in atretic small antral follicles.

Article Snippet: The primary antibodies used were mouse anti-CYP19A1 (1:100, sc-374176, Santa Cruz, Dallas, TX, USA), mouse anti-AR (1:100, sc-7305, Santa Cruz), mouse anti-AMH (1:100, MCA2246T, BioRad, Hercules, CA, USA), rabbit anti-CDH1 (1:100, CST3195S, Cell Signaling, Danvers, MA, USA), rabbit anti-MKI67 (1:100, ab15580, Abcam, Cambridge, UK), rabbit anti-cCASP3 (Asp175) (1:100, CST9661S, Cell Signaling), rabbit anti-COLLIV (1:100, AB748, Merck), mouse anti-CD68 (1:100, M087629-2, DAKO, Glostrup, Denmark), rabbit anti-ACTA2 (1:100, ab5694, Abcam), mouse anti-STAR (D-2) (1:100, sc-166821, Santa Cruz), rabbit anti-GSTA1 (1:100, HPA053817, Atlas Antibodies, Bromma, Sweden).

Techniques: Expressing, TUNEL Assay

Journal: Nature Communications

Article Title: Single-cell reconstruction of follicular remodeling in the human adult ovary

doi: 10.1038/s41467-019-11036-9

Figure Lengend Snippet: Complement and immune system in the adult ovaries. a tSNE cluster map showing expression of selected complement genes. b Gene network of C1S in TC and stroma. The color of the circles represents function and the color of the edges represent networks. c Immunostaining of follicles (ø, diameter) growing (top row), atretic (middle row), and degenerated (bottom row) for C1S, C1Q, and CD68. Single channel images were converted to an intensity map. Scale bars are 100 μm. d Immunostaining of follicles (ø, diameter) growing (top row), atretic (middle row), and degenerated (bottom row) for C1Q and STAR. Scale bars are 100 μm. e Concentration of secreted C1Q and C3 produced by pieces of human ovarian stroma ( N = 3 samples) and atretic follicle walls ( N = 6 samples) after 1 and 5 days of culture. Median and sample distribution (dots) are shown. f tSNE cluster map showing expression of C1QA and C3 . g–i tSNE cluster map showing expression of selected immune marker genes for innate immune cells g , B lymphocytes h and T lymphocytes and NK cells i . Red dashed lines give the boundaries of the specific immune-clusters

Article Snippet: The primary antibodies used were rabbit anti-KI67 (1:100, ab15580, Abcam), mouse anti-AMH (1:30, MCA2246T, BioRad), rabbit anti-Troponin I (H-170) (1:100, sc15368, Santa Cruz), goat anti-VASA/DDX4 (1:200, AF2030, R&D), mouse anti-Cytokeratin (1:100, M351501, DAKO), rabbit anti-Wilm’s Tumor protein (1:100, CA1026-50, Calbiochem), mouse anti-StAR (D-2) (1:100, sc166821, Santa Cruz), rabbit anti-alpha smooth muscle actin/ACTA2 (1:200; ab5694, Abcam), goat anti-ZP3 (N-20) (1:100, sc23715, Santa Cruz), mouse anti-Connexin-43/GJA1 (CX-1B1) (1:50, 13-8300, Zymed), rabbit anti-Fragilis/IFITM3 (1:200, ab15592, Abcam), rabbit anti-Collagen Type IV (1:50, AB748, Chemicon), rabbit anti-Von Willebrand factor/VWF (1:100, ab6994, Abcam), mouse anti-N-Cadherin/CDH2 (GC-4) (1:100, C3865, Sigma-Aldrich), rabbit anti-PECAM1 (M-20) (1:200, sc1506, Santa Cruz), rabbit anti-c-FOS (1:20, PC38, Calbiochem), mouse anti-8-OHdG (1:1000, sc66036, Santa Cruz), goat anti-IGFBP5 (1:50, AF875, R&D), rabbit anti-C1Q (1:400, A0136, DAKO), goat anti-C1S (1:400, A302, Quidel) and mouse anti-CD68 (1:50, M087629-2, DAKO).

Techniques: Expressing, Immunostaining, Concentration Assay, Produced, Marker