Apoptosis Occurs in Granulosa Cells but not Cumulus Cells in the Atretic Graafian Follicles in Multiparous Pig Ovaries.

Manabe, Noboru; Imai, Yuzuru; Myoumoto, Akira; Kimura, Yuichi; Sugimoto, Miki; Okamura, Yosuke; Fukumoto, Manabu; Sakamaki, Kazuhiro; Miyamoto, Hajime

doi:10.1267/ahc.30.85

Cited by 24 publications

(42 citation statements)

References 34 publications

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“…As described in our previous reports [6][7][8][9][10], oocytes were tightly surrounded by cumulus cells in healthy follicles of porcine ovaries, and they were directly adjacent to granulosa cell layers without degenerating cells with apoptotic bodies, a morphological hallmark of apoptosis, and no degenerating cells were seen in the theca interna or externa layers (Figs. 1B, 2B and 3B).…”

Section: Histopathologysupporting

confidence: 55%

“…Ovarian sections were stained by the TUNEL method using a commercial kit (Apop Tag; Intergen, Purchase, NY, USA) according to a modification of the method reported previously [6][7][8][9][10] to allow visualization of the apoptotic cells. Briefly, the sections were fixed in 10% neutralbuffered formalin (pH.…”

Section: Histochemistrymentioning

confidence: 99%

“…As negative controls, the sections were incubated with omission of either TdT or DIG-ddUTP. As a positive control, tissue sections were treated with DNase I (1 µg/ml; Boehringer Mannheim, Indianapolis, IN, USA), 140 mM sodium cacodylate, 4 mM MgCl 2 and 0.1 mM dithiothreitol in 30 mM Tris-HCl, pH 7.2, for 10 min at RT before exposure to TdT, and sections prepared from young adult rat testes were used as physiological positive controls [6][7][8][9][10].…”

Section: Histochemistrymentioning

confidence: 99%

“…Ovarian follicular atresia is affected by various factors such as steroids, gonadotrophins and cytokines, but the mechanism that regulates follicle selection during follicular development is not well understood [6][7][8]. Our previous studies of follicular atresia in porcine ovaries demonstrated that degeneration of the atretic follicles can be explained partly by apoptosis of granulosa cells [9,10].…”

mentioning

confidence: 99%

“…In the present study, the expression of classic cadherins and β-catenin, which is associated with the membrane-anchored cytoplasmic domain of classic cadherins [13], in developing and/or atretic follicles of porcine ovaries was investigated by immunocytochemical analysis. The principal aims were: first, to determine what types of cadherins are expressed in porcine follicles; second, to investigate the pattern of expression in follicles during follicular atresia, especially in granulosa cells, apoptotic cell death of which is considered to be a key phenomenon in follicular atresia and which contributes to determination of the follicle population in each estrous cycle [6][7][8][9][10]; and third, to determine whether there are any differences in expression of cadherins in differentiating follicular cells.…”

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confidence: 99%

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Changes in Cell Adhesion Molecules during Follicular Atresia in Porcine Ovaries.

Nishihara

Manabe

Nakayama

et al. 2000

Journal of Reproduction and Development

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Abstract. Determination of the expression level and localization of cell adhesion molecules is crucial for understanding the mechanism of maintenance and remodeling of the ovarian follicle structure. We immunocytochemically investigated expression of the cell adhesion molecules, "classic" cadherins and β-catenin, in developing and/or atretic follicles of porcine ovaries. Healthy follicles showed strong staining for cadherin-8 and β-catenin in granulosa cells tightly attached to the basement membrane, and moderate/weak staining was seen on the inner surface of the granulosa cell layer. Strong VE-cadherin expression was seen in a single cell layer attached to the basement membrane in the theca interna layer of healthy follicles. The expression of cadherin-8, β-catenin and VE-cadherin decreased during follicular atresia. No positive staining was observed for R-cadherin, E-cadherin, Tcadherin, BR-cadherin or P-cadherin, and a weak positive reaction for N-cadherin was seen only in the granulosa cells of healthy and early atretic follicles. These findings indicate that cadherin-8, Ncadherin and VE-cadherin have important roles in follicular development and/or degeneration, and that decreases in the expression of these cadherins are involved in follicular atresia in porcine follicles.

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Section: Histopathologysupporting

confidence: 55%

Section: Histochemistrymentioning

confidence: 99%

Section: Histochemistrymentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Changes in Cell Adhesion Molecules during Follicular Atresia in Porcine Ovaries.

Nishihara

Manabe

Nakayama

et al. 2000

Journal of Reproduction and Development

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Regulatory Mechanisms of Granulosa Cell Apoptosis in Ovarian Follicle Atresia

Manabe

Kimura

Uchio

et al.

Animal Cell Technology: Challenges for the 21st Century

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Porcine ovary samples were prepared for histochemical and ultrastructural analyses. In situ analysis of DNA fragmentation was performed on histological sections of follieles using the terminal deoxynucleotidyl transferase-mediated biotinylated deoxyuridine triphosphate nick end-labeling TUNEL) method. No apoptotic cells were observed in healthy follicles. In atretic follicles, apoptotic TUNEL staining was seen in scattered granulosa cells located on the inner surface of the follicular wall, but not in cumulus cells, internal or external theca cells, or oocytes. Nuclear condensation, a typical feature of apoptosis was seen only in scattered granulosa cells. The neutral Ca² + /Mg² +dependent endonuclease is involved in granulosa cell apoptosis. No endonuclease activity was detected in cumulus cells. An IgM monoclonal antibody (PFG-1) capable of inducing granulosa cell apoptosis was then produced against granulosa cells prepared from healthy antral follicles. Two-dimensional (2D) Western blotting analysis revealed that PFG-1 specifically recognized a cell-membrane protein (PFG-1 antigen, 55 kD, pl 5.9). PFG-1 immunohistochemically reacted with granulosa cells of healthy follicles but not those of atretic follicles. When the isolated granulosa cells prepared from healthy follicles were cultured in medium containing 0.1 µg/ml of PFG-1, the cells underwent apoptosis. These observations indicated that apoptosis occurs in granulosa cells but not cumulus cells in the atretic follicles and that the PFG-1 antigen, a novel cell death receptor, is different from the apoptosis-mediating receptors Fas antigen or tumor neerosis factor receptor 1 (TNFR-1).

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Follicle Selection in Mammalian Ovaries: Regulatory Mechanisms of Granulosa Cell Apoptosis during Follicular Atresia

Manabe¹,

Inoue²,

Sugimoto³

et al. 2004

The Ovary

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Apoptosis Occurs in Granulosa Cells but not Cumulus Cells in the Atretic Graafian Follicles in Multiparous Pig Ovaries.

Cited by 24 publications

References 34 publications

Changes in Cell Adhesion Molecules during Follicular Atresia in Porcine Ovaries.

Changes in Cell Adhesion Molecules during Follicular Atresia in Porcine Ovaries.

Regulatory Mechanisms of Granulosa Cell Apoptosis in Ovarian Follicle Atresia

Follicle Selection in Mammalian Ovaries: Regulatory Mechanisms of Granulosa Cell Apoptosis during Follicular Atresia

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