• Title/Summary/Keyword: Granulosa cells

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Apoptosis and Expression of Fas nnd Fas Lignnd Genes in Mouse Ovary (생쥐 난소내 Fas 및 Fas Ligand의 발현과 세포자연사)

  • Yoon Jeong-Mi;Yoon Hyunsook;Yang Hyunwon;Kim Seikwang;Cho Dong Jae;Yoon Yong-Dal
    • Development and Reproduction
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    • v.7 no.1
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    • pp.15-22
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    • 2003
  • The Fas antigen (Fas) as a cell-surface receptor protein which mediates apoptosis-inducing signals plays an important role in the immune system. Expression of Fas mRNA is detected not only in lymphoid organs but also in the nonlymphoid organs. In the ovary, most of the follicles is known to undergo atreisa through apoptosis. However, the exact mechanism of atresia was not elucidated yet. Therefore, the purposes of the present study were to investigate the expression of Fas and Fas ligand in mouse ovary and to clarify the relationship between expression of Fas and Fas ligand and atresia of follicle. The result of RT-PCR demonstrated that Fas and Fas ligand mRNA was expressed in ovary, especially granulosa cells and oocytes. The immunohistochemistry showed that the granulosa cells and oocytes in growing follicles were stained for Fas and Fas ligand, but primordial follicles were not. Furthermore, Fas and Fas ligand were intensively stained in the atretic follicles As results of TUNEL staining to detect apoptotic cells in the ovaries, the number of TUNEL-positive (apoptotic) granulosa cells and oocytes increased in the atretic follicles compared to the healthy normal follicles. These results demonstrate that there is the positive relationship between expression of Fas and Fas ligand in granulosa cells and oocyies and apoptosis of them leading to atresia of follicles. It suggests that expression of Fas and Fas ligand could be associated with atresia of follicles in mouse ovary.

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The potential role of granulosa cells in the maturation rate of immature human oocytes and embryo development: A co-culture study

  • Jahromi, Bahia Namavar;Mosallanezhad, Zahra;Matloob, Najmeh;Davari, Maryam;Ghobadifar, Mohamed Amin
    • Clinical and Experimental Reproductive Medicine
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    • v.42 no.3
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    • pp.111-117
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    • 2015
  • Objective: In order to increase the number of mature oocytes usable for intracytoplasmic sperm injection (ICSI), we aimed to investigate the effect of co-culturing granulosa cells (GCs) on human oocyte maturation in vitro, the fertilization rate, and embryo development. Methods: A total of 133 immature oocytes were retrieved and were randomly divided into two groups; oocytes that were cultured with GCs (group A) and oocytes that were cultured without GCs (group B). After in vitro maturation, only oocytes that displayed metaphase II (MII) underwent the ICSI procedure. The maturation and fertilization rates were analyzed, as well as the frequency of embryo development. Results: The mean age of the patients, their basal levels of follicle-stimulating hormone, and the number of oocytes recovered from the patients were all comparable between the two study groups. The number of oocytes that reached MII (mature oocytes) was 59 out of 70 (84.28%) in group A, compared to 41 out of 63 (65.07%) in group B (p=0.011). No significant difference between fertilization rates was found between the two study groups (p=0.702). The embryo development rate was higher in group A (33/59, 75%) than in group B (12/41, 42.85%; p=0.006). The proportion of highest-quality embryos and the blastocyst formation rate were significantly lower in group B than in group A (p=0.003 and p<0.001, respectively). Conclusion: The findings of the current study demonstrate that culturing immature human oocytes with GCs prior to ICSI improves the maturation rate and the likelihood of embryo development.

Expression of the genes for peroxisome proliferator-activated receptor-γ, cyclooxygenase-2, and proinflammatory cytokines in granulosa cells from women with polycystic ovary syndrome

  • Lee, Joong Yeup;Tae, Jin Cheol;Kim, Chung Hyon;Hwang, Doyeong;Kim, Ki Chul;Suh, Chang Suk;Kim, Seok Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.44 no.3
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    • pp.146-151
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    • 2017
  • Objective: To identify differences in the expression of the genes for peroxisome proliferator-activated receptor $(PPAR)-{\gamma}$, cyclooxygenase (COX)-2, and the proinflammatory cytokines interleukin (IL)-6 and tumor necrosis factor $(TNF)-{\alpha}$ in granulosa cells (GCs) from polycystic ovary syndrome (PCOS) patients and controls undergoing controlled ovarian stimulation. Methods: Nine patients with PCOS and six controls were enrolled in this study. On the day of oocyte retrieval, GCs were collected from pooled follicular fluid. Total mRNA was extracted from GCs. Reverse transcription was performed and gene expression levels were quantified by realtime quantitative polymerase chain reaction. Results: There were no significant differences in age, body mass index, and total gonadotropin dose, except for the ratio of luteinizing hormone to follicle-stimulating hormone between the PCOS and control groups. $PPAR-{\gamma}$ and COX-2 mRNA was significantly downregulated in the GCs of PCOS women compared with controls (p= 0.034 and p= 0.018, respectively), but the expression of IL-6 and $TNF-{\alpha}$ mRNA did not show significant differences. No significant correlation was detected between the expression of these mRNA sequences and clinical characteristics, including the number of retrieved oocytes, oocyte maturity, cleavage, or the good embryo rate. Positive correlations were found among the $PPAR-{\gamma}$, COX-2, IL-6, and $TNF-{\alpha}$ mRNA levels. Conclusion: Our data may provide novel clues regarding ovarian GC dysfunction in PCOS, and indirectly provide evidence that the effect of $PPAR-{\gamma}$ agonists in PCOS might result from alterations in the ovarian follicular environment. Further studies with a larger sample size are required to confirm these proposals.

Expression of Proapoptotic Bcl-2 Family Member in the Mouse Ovary (I) (생쥐 난소에서 Bcl-2계 세포고사인자에 관한 연구 (I))

  • Lee, Yu-Il;Lee, Jin;Chun, Sang-Young
    • Clinical and Experimental Reproductive Medicine
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    • v.30 no.1
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    • pp.47-55
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    • 2003
  • Objectives: Bok, Bcl-2-related ovarian killer, is a proapoptotic Bcl-2 family protein identified in the ovary based on its dimerization with the antiapoptotic protein Mcl-1. The present study examined the hormonal regulation and localization of Bok messenger RNA levels in the mouse ovary during the follicle development. Methods: The animals were implanted subcutaneously with Silastic brand capsules containing the synthetic estrogen, DES at $21{\sim}23$ days of age. Ovaries were collected $1{\sim}3$ days after implantation for RNA analysis and in situ hybridization. Some mice were removed capsule for $1{\sim}2$ days to induce ovarian follicle apoptosis. Ovaries were also collected from 26 day-old immature mice at various times after treatment with 10 IU PMSG. Some mice received a single intraperitoneal injection of 10 IU hCG to induce ovulation, and ovaries were obtained at different time intervals for Northern blot and in situ hybridization analysis, respectively. Results: Treatment of immature mice with diethylstilbestrol (DES) for $24{\sim}48$ h increased ovarian Bok mRNA levels. Bok mRNA was remained the same levels in mice removed DES for $24{\sim}48$ h to induce apoptosis. High signals of Bok mRNA after DES treatment were detected in granulosa cells of early antral follicles. Treatment of immature mice with PMSG for 12 h increased markedly ovarian Bok mRNA expression which was detected mainly in preantral and atretic follicles. Interestingly, low levels of Bok mRNA were also expressed in granulosa cells of preovulatory follicles. Treatment of PMSGprimed mice with hCG stimulated strongly ovarian Bok mRNA expression at $6{\sim}9$ h. At that time, Bok mRNA was expressed in granulosa cells of atretic and small growing follicles. Conclusion: These results demonstrate that Bok is one of proapoptotic Bcl-2 members expressed in early growing and atretic follicles during the ovarian follicular development. Gonadotropins induce a transient increase of Bok gene expression in granulosa cells of preantral and preovulatory follicles indicating some role in the ovulatory process.