생쥐 난소에서 Bcl-2계 세포고사인자에 관한 연구 (I)

Expression of Proapoptotic Bcl-2 Family Member in the Mouse Ovary (I)

  • 이여일 (전남대학교 의과대학 산부인과학교실) ;
  • 이진 (전남대학교 의과대학 산부인과학교실) ;
  • 전상영 (전남대학교 의과대학 호르몬연구센터)
  • Lee, Yu-Il (Department of Obstetrics and Gynecology, College of Medicine, Chonnam National University) ;
  • Lee, Jin (Department of Obstetrics and Gynecology, College of Medicine, Chonnam National University) ;
  • Chun, Sang-Young (Hormone Research Center, Chonnam National University)
  • 발행 : 2003.03.30

초록

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.

키워드

참고문헌

  1. Kerr JF, Wyllie AH, Currie AR. Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 1972; 26: 239-57 https://doi.org/10.1038/bjc.1972.33
  2. Hsu SY, Lai RJ, Finegold M, Hsueh AJW. Targeted overexpression of Bcl-2 in ovaries of transgenic mice leads to decreased follicle apoptosis, enhanced folliculogenesis and increased tumorigenesis. Endocrinology 1996; 137: 4837-43 https://doi.org/10.1210/en.137.11.4837
  3. Hsu SY, Hsueh AJW. Intracellular mechaniams of ovarian cell apoptosis. Mol Cell Endocrinol 1998;145: 21-5 https://doi.org/10.1016/S0303-7207(98)00165-8
  4. Oltvai ZN, Milliman CL, Korsmeyer SL. Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell 1993; 74: 609-19 https://doi.org/10.1016/0092-8674(93)90509-O
  5. Hsu SY, Hsueh AJW. Tissue-specific Bcl-2 protein partners in apoptoisis: an ovarian paradigm. Physiol Rev 2000; 80: 593-614 https://doi.org/10.1152/physrev.2000.80.2.593
  6. Hsu SY, Kaipia A, McGee E, Lomeli M, Hsueh AJW. Bok is a pro-apoptotic Bcl-2 protein with restricted expression in reproductive tissues and heterodimerizes with selective anti-apoptotic Bcl-2 family members. Proc Natl Acad Sci USA 1997; 94:12401-6 https://doi.org/10.1073/pnas.94.23.12401
  7. Adams JM, Cory S. The Bcl-2 protein family: arbiters of cell survival. Science 1998; 281: 1322-6 https://doi.org/10.1126/science.281.5381.1322
  8. Suominen JS, Yan W, Toppari J, Kaipia A. The expression and regulation of Bcl-2-related ovarian killer (Bok) mRNA in the developing and adult rat testis. Euro J Endocrinol 2001; 145: 771-8 https://doi.org/10.1530/eje.0.1450771
  9. Ha SH, Lee SR, Lee TH, Kim YM, Baik MG, Choi YJ. The expression of Bok is regulated by serum in HC11 mammary epithelial cells. Mol Cells 2001; 12: 368-71
  10. Lee J, Park HJ, Choi HS, Kwon HB, Arimura A, Lee BJ, et al. Gonadotropin stimulation of pituitary adenylate cyclase-activating polypeptide (PACAP) messenger ribonucleic acid in the rat ovary and the role of PACAP as a follicle survival factor. Endocrinology 1999; 140: 818-26 https://doi.org/10.1210/en.140.2.818
  11. Park JY, Park JH, Park HJ, Lee JY, Lee YI, Lee K, et al. Stage-dependent regulation of ovarian pitui- tary adenylate cyclase-activating polypeptide mRNA levels by GnRH in cultured rat granulosa cells. Endocrinology 2001; 142: 3828-35 https://doi.org/10.1210/en.142.9.3828
  12. Billig H, Furuta I, Hsueh AJW. Estrogens inhibit and androgens enhance ovarian granulosa cell apoptosis. Endocrinology 1993; 133: 2204-12 https://doi.org/10.1210/en.133.5.2204
  13. Hsueh AJW, Elsenhauer SY, Chun SY, Hsu SY, Billig H. Gonadal cell apoptosis. Recent Prog Horm Res 1996; 51: 433-55
  14. Leo CP, Hsu SY, Chun SY, Bae HW, Hsueh AJW. Characterization of the antiapoptotic Bcl-2 family member myeloid cell leukemia-1 (Mcl-1) and the stimulation of its message by gonadotropin in the rat ovary. Endocrinology 1999; 140: 5469-77 https://doi.org/10.1210/en.140.12.5469
  15. Chun SY, Bae HW, Kim WJ, Park JH, Hsu SY, Hsueh AJW. Expression of essenger ribonucleic acid for the antiapoptosis gene P11 in the rat ovary: gonadotropin stimulation in granulosa cells of preovulatory follicles. Endocrinology 2001; 142: 2311-7 https://doi.org/10.1210/en.142.6.2311
  16. Inohara N, Ekhterae D, Garcia I, Carrio R, Merino J, Merry A. Mtd, a novel Bcl-2 family member activates apoptosis in the absence of heterodimerization with Bcl-2 and Bcl-XL. J Biol Chem 1998; 273: 8705-10 https://doi.org/10.1074/jbc.273.15.8705
  17. Chun SY, Hsueh AJW. Paracrine mechanisms of ovarian follicle apoptosis. J Reprod Immunol 1998; 39: 63-75 https://doi.org/10.1016/S0165-0378(98)00013-8
  18. Richards JS. Hormonal control of gene expression in the ovary. Endocr Rev 1994; 15: 725-51 https://doi.org/10.1210/edrv-15-6-725
  19. Wong WYL, DeWitt DL, Smith WL, Richards JS. Rapid induction of prostagladin endoperoxide synthase in rat preovulatory follicles by luteinizing hormone and cAMP is blocked by inhibitors of transcription and translation. Mol Endocrinol 1989;3: 1714-23 https://doi.org/10.1210/mend-3-11-1714
  20. Galway AB, Lapolt PS, Tsafriri A, Dargan CM, Boime I, Hsueh AJW. Recombinant FSH induces ovulation and tissue plasminogen activator expres-sion in hypophysectomized rats. Endocrinology 1990; 127: 3023-8 https://doi.org/10.1210/endo-127-6-3023
  21. Natraj U, Richards JS. Hormonal regulation, localization, and functional activity of the progesterone receptor in granulosa cells of rat preovulatory follicles. Endocrinology 1993; 133: 761-9 https://doi.org/10.1210/en.133.2.761
  22. Braw RH, Bar-Ami S, Tsafriri A. Effect of hypophysectomy on atresia of rat preovulatory follicles. Biol Reprod 1981; 25: 989-96 https://doi.org/10.1095/biolreprod25.5.989