• Development and Reproduction
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• v.1 no.2
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• pp.157-164
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• 1997

To study the regulation of porcine follicular cell apostosis by gonadotropin, steroid, and nitric oxide, we analyzed DNA fragmentation, the hallmark of apoptosis, and nitrite production of porcine granulosa cells. Dissected indiidual follicles from ovary were separated in size (small, 2-3 mm; medium, 5-6 mm; large, 7-8 mm) and isolated granulosa cells were classified morpholocally as atretic or nonatretic. Nitrite concentration was measured by mixing follicular fluids with an equal volume of Griess reagent. Follicular nitric oxide (NO) concentration of healthy follicles was higher than that of atretic follicles. Apoptotic DNA fragmentation was suppressed in non-apoptotic granulosa cells. Follicular apoptosis was induced by androgen but prevented by gonadotropin in vitro. Apoptosis was confined to the granulosa cells. But it was not clear whether apoptosis of granulosa cells were isolated, incubated with or without gonadotropin, androgen and sodium nitroprusside (SNP), respectively at $37^{\circ}C$ for 24 hrs. Cultured granulosa cells were used to extract genomic DNA and culture media was asssayed for nitrite concentration. Nitrite production of culture media was increased, while apoptotic DNA fragmentation was suppressed in PMSG, hCG, testosterone+SNP and SNP treated groups. Nitrite concentration in culture media was decreased, but apoptotic DNA fragmentation was induced in testosterone treated group. These data suggest that NO production and apoptosis may be involved of granulosa cell apoptosis induced by testosterone.

• Proceedings of the Zoological Society Korea Conference
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• 1994.10a
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• pp.161.2-161
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• 1994

No Abstract, See Full Text

• Korean Journal of Veterinary Research
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• v.48 no.3
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• pp.323-326
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• 2008

A two-year-old mixed breed sow was requested to the Veterinary Pathology Laboratory of Cheju National University with a clinical signs of severe abdominal pain and sudden death. Grossly, there was severe hemorrhage in abdominal cavity. Most of internal parenchymas and subcutaneous muscle showed severe pale discoloration. Both ovaries were enlarged with oval to round protruding multilobular masses and dark red in color. And they were firm and contained multiple small cysts in their cut surface. Histopathologically, numerous neoplastic granulosa cells had spherical-to-oval, hyperchromatic nuclei and scant eosinophilic cytoplasms were distributed with follicular pattern in ovarian masses. And the typical Call-Exner bodies, distinctive microcavityies, were observed in the center of small neoplastic follicles. Based on the gross and histopathologic findings, this case was diagnosed as granulosa cell tumor. In our best knowledge, this is believed to be the first report of granulosa cell tumor in a sow in Korea.

• Korean Journal of Veterinary Service
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• v.47 no.3
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• pp.123-132
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• 2024

The stressful conditions including heat stress negatively affect to the animal's reproductive performance. Apoptosis of granulosa cells under unexpected stimuli can influence the fate of ovarian follicles and its function. Since the changes in caspase-3, a key executioner of the apoptosis pathway, have not yet been elucidated in heat-stressed sows, this study compared its changes in the ovaries of domestic sows affected by heat stress during summer. The samples were collected at spring (23.1℃; CON group) as suitable temperature for raising pigs or at summer (34.4℃; HS group) as unavoidable chronic heat stress. It was not significant but strong tendency to release higher level of stress hormone (corticosterone) in HS group than that in CON group was identified. A significant upregulation of cleaved (activated) caspase-3 expression in HS group was identified compared to CON group, indicating that apoptosis was increased in the ovarian follicles during summer. The expression of cleaved caspase-3 in the ovaries was positively related with degree of maximal atmospheric temperature and serum levels of corticosterone. The caspase-3-positive cells were more prevalent in mature follicles with localization primarily in the granulosa cell layer than immature follicles. The intensities of caspase-3 in the granulosa cell layer were stronger in HS group than CON group. In conclusion, the present study demonstrated that chronic heat stress during summer in Korea increased activated caspase-3-related apoptosis in the granulosa cells in the mature ovarian follicles, which might suggest that heat stress decreased reproductive performance in the domestic sows during summer.

• Clinical and Experimental Reproductive Medicine
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• v.29 no.3
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• pp.195-200
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• 2002

Objective : To investigate the expression of apoptosis related proteins and apoptotic cells on the human ovarian follicles. Materials and Methods: Thirty five Formalin-fixed paraffin-embedded human ovarian tissue blocks were selected from the surgical pathology files of the department of pathology, College of Medicine, Yonsei University, for the period from 1996 to 1998. All specimen were from premenopausal women aged from $32{\sim}45$. Ovarian tissues were collected from the patients performing hysterectomy for benign uterine diseases. Immunohistochemical staining was performed for the detection of DNA fragmented cell, Bcl-2, Bax, Fas and Fas-ligand. Results: Bcl-2 and bax were not expressed on the surrounding cells and oocyte of the primary, primordial and preantral follicles. Fas and Fas-ligand (Fas-L) were not expressed on the surrounding cells on the primordial and primary follicles. But expressed on the surrounding granulosa cells and oocyte in the primordial and primary follicles. In the healthy follicles, Bcl-2 was expressed on the granulosa cells, however, Bax was not expressed. DNA fragmented cells were expressed on the inner granulosa cell layer of atretic follicles. Conclusion: Fas, Fas-ligand, and Bax may be responsible for the follicular atresia and Bcl-2 may be involved in the follicular survival in the human ovary.

• Korean Journal of Veterinary Service
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• v.46 no.3
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• pp.227-234
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• 2023

It has been addressed that heat stress due to high atmospheric temperature during summer in Korea induces impaired release of reproductive hormones, followed by occurring abnormal ovarian cyclicity, lower pregnancy ratio, and reduced litter size. Therefore, the present study attempted to compare seasonal change (spring versus summer) of the ovarian aromatase expression, an enzyme for converting testosterone into estrogen. While serum estrogen level in summer group was significantly lower than that of spring group, testosterone was not different between groups. Consistent with estrogen level, the ovarian aromatase expression in summer at follicular phase was significantly lower than the counterpart of spring. The ovarian aromatase expression was positively related with serum estrogen level significantly (r=0.689; P=0.008) and strongly negative correlation was identified (r=-0.533; P=0.078) with atmospheric temperature. The ovarian aromatase expression was not detected in immature ovarian follicles but specifically localized in the granulosa cell layers in both seasons. However, the aromatase intensity in the granulosa cell layers was stronger in spring than summer. Because testosterone level was not different between groups, it could be concluded that the lower level of estrogen during summer might be derived by not lack of substrate but lower expression of ovarian aromatase by heat stress.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.3
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• pp.1351-1355
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• 2014

Background: Ovarian sex-cord stromal tumours (SCST) are rare, and relatively infrequent in children. These have to be distinguished from more common germ cell tumors in children and also from benign epithelial neoplasms. Objectives: The purpose of our study was to report the clinical and pathological findings in young patients with these tumours in our population. Material and Methods: The present observational cross-sectional study included all subjects <21 years of age diagnosed with ovarian SCST, in Aga Khan University Hospital Histopathology Laboratory, Karachi, Pakistan, from January 1992 till July 2013. Results: Of the total of 513 SCSTs presented during the study period, 39 fulfilled inclusion criteria and were assessed. The age range was 4-250 months. Most of the tumours presented at stage-1 and an abdominal mass was the most common presenting symptom, along with menstrual disturbance. The left side ovary was slightly more affected (53.5%). Of the total, 15 were juvenile granulosa cell tumours (JGCT), 11 sclerosing stromal tumours (SST), 10 of the fibrothecomas spectrum, 2 Sertoli leydig cell tumours (SLCT) and one a sex cord tumour with annular tubules (SCTAT). Detailed immunohistochemical analyses were performed in 33 cases. Recurrence/metastasis was noted in 4/21 cases with follow-up data. Conclusions: Ovarian sex cord stromal tumours are very rare in young age in our population, and usually present at an early stage. Most common among these are juvenile granulosa cell tumours, although surprisingly sclerosing stromal tumours were also common. Clinical symptoms due to hormone secretion in premenstrual girls and menstrual disturbance in menstruating girls are common presenting features.

• Korean Journal of Clinical Laboratory Science
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• v.38 no.1
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• pp.72-81
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• 2006

Wee1 is a kinase regulator of the M-phase promoting factor (MPF; a complex of cdc2 and cyclin B1). The present study was undertaken to determine the role(s) of wee1 in the early stages of mouse ovarian follicles. The expression of wee1 and the correlated cell-cycle components, namely cdc2, cyclin B1, and cdc25C, were evaluated by immunohistochemistry. In addition, the expression of Tyr15-phosphorylated cdc2 (cdc2-p) was also examined to determine whether wee1 kinase phosphorylates cdc2 existed. Each component except cdc25C was found cytoplasmic in the oocytes at all stages of follicles, while cdc25C was not detected in primordial follicles. It was found primarily in ovarian somatic cells and to a small extent in granulosa cells of the growing follicles. To further confirm the expression of cell-cycle components in the primordial follicular oocytes, day1 ovaries were enzymatically and mechanically dissociated, then oocytes were isolated from somatic including pre-granulosa cells, and we confirmed that cdc2-p was expressed in oocytes of primordial follicles. From the results of the present study, we concluded wee1, without the counteracting cdc25C, would cause meiotic arrest of oocytes by the inhibitory phosphorylation of cdc2. The expression of all these proteins in the granulosa cells of growing follicles may regulate their mitosis concurrently with the growth of oocytes and follicles.

• Clinical and Experimental Reproductive Medicine
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• v.20 no.2
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• pp.137-147
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• 1993

The purpose of this study is to identify the presence of the circulating antibodies directed toward ovarian proteins{antiovarian antibodies, AOA) and the nature of antigenic ovarian structure by comparing the binding activities to 4 types of ovarian proteins, particulated and solubilized forms of pig ovarian and granulosa cell membranes in sera of patients with premature ovarian failure(POF) and to evaluate the usefulness of circulating AOA as a follow up tool after treatment. Measurements of AOA were performed by enzyme linked immunosorbent assay(ELISA) in sera of 58 patients with POF, 51 had normal chromosomes and 7 had X chromosome abnormalities. Sera of 21 natural menopausal women and 17 castrated women were also tested and sera of 32 healthy premenopausal women were served as controls. ELISA reactivities against particulated porcine granulosa cell membrane proteins was the greatest among 4 different ovarian proteins. Fifteen(29%) of 51 POF patients with normal chromosome and 1(14.3%) of 7 POF patients with X chromosome abnormalities had AOA while none of 32 controls and 21 natural menopausal women and 17 castrated women had AOA. One POF patient with 47, XXX was identified AOA positive. The ELISA reactivities were followed up monthly up to 5 months in 4 AOA positive POF patients after estrogen-progestin{E-P) therapy. There was a decreasing tendency of the ELISA reactivities in all these patients after E-P therapy and two of them converted to AOA negative. These data suggest that antigenic structure may be components of granulosa cell membrane and the determination of circulating AOA may be useful in the follow up after treatment in patients with autoimmune POF.

• Clinical and Experimental Reproductive Medicine
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• v.26 no.1
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• pp.55-65
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• 1999

There have been many reports to date regarding the role of GnRH as a local regulatory factor of ovarian function as studies of human and rat ovaries revealed GnRH and its receptor. In recent studies it has been shown that GnRH directly causes apoptosis in the granulosa cells of the rat ovary, and such results leads to the suggestion that the use of GnRH agonist for more stable long term ovarian hyperstimulation in human IVF-ET programs causes granulosa cell apoptosis which may lead to follicular atresia. Therefore this study attempts to determine if granulosa-luteal cell apoptosis occurs in patients during IVF-ET programs in which GnRH agonist is employed for ovarian hyperstimulation. The quality of oocyte-cumulus complexes obtained during ovum pickup procedures were assessed morphologically and then the fertilization rate and developmental rate was determined. Apoptotic cells among the granulosa-luteal cells obtained during the same procedure were observed after staining with Hematoxylin-eosin. The fragmentation degree of DNA extracted from granulosa-luteal cells was determined and comparatively analyzed. There was no difference in the average age of the patients, the number of oocytes retrieved, and fertilization and developmental rates between the FSH/hMG group and GnRH-long group. There was also no difference in the apoptosis rate and pyknosis rate in the granulosa-luteal cells between the two groups. However, when the oocyte-cumulus complexes were morphoogically divided into the healthy group and atretic group without regard for the method of hyperstimulation, the results showed that the number of oocytes obtained averaged $11.09{\pm}8.75\;and\;10.33{\pm}4.53$ per cycle, respectively, showing no significant difference, but the fertilization rate (77.05%, 56.99%, respectively, p<0.01) and developmental rate (65.96%, 41.51%, respectively, p<0.01) was significantly increased in the healthy group when compared to the atretic group. The degree of apoptosis in the granulosa-luteal cells showed that in the healthy group it was 2.25% which was not significantly different from the atretic group (2.77%), but the pyknosis rate in the atretic group (27.81%) was significantly higher compared to the healthy group (11.35%, p<0.01). The quantity of DNA fragmentation in the FSH/hMG group was 32.22%, while in the GnRH-long group it was 34.27%, showing no significant difference. On the other hand the degree of DNA fragmentation was 39.05% and 11.83% in the healthy group and atretic group, respectively, showing significantly higher increase in the atretic group (p<0.01). The above results suggest that death of granulosa-luteal cells according to the state of the oocyte-cumulus complex is more related to pyknosis rather than apoptosis. Also, the GnRH agonist used in ovarian hyperstimulation does not seem to directly affect the apoptosis of retrieved oocytes and granulosa-luteal cells, and which is thought to be due to the suppression of the apoptogenic effect of GnRH agonist as a result of the high doses of FSH administered.