Control Mechanisms of Ovarian Follicle Development by Follicle Stimulating Hormone and Pituitary Adenylate Cyclase-activating Polypeptide

난포자극호르몬과 Pituitary Adenylate Cyclase-activating Polypeptide에 의한 난소의 난포성장

  • Lee, Yu-Il (Department of Obstetrics and Gynecology, Medical School, Chonnam National University) ;
  • Shin, Jin-Ok (Department of Obstetrics and Gynecology, Medical School, Chonnam National University) ;
  • Kim, Mi-Young (Department of Obstetrics and Gynecology, Medical School, Chonnam National University) ;
  • Chun, Sang-Young (Hormone Research Center, Chonnam National University)
  • 이여일 (전남대학교 의과대학 산부인과학교실) ;
  • 신진옥 (전남대학교 의과대학 산부인과학교실) ;
  • 김미영 (전남대학교 의과대학 산부인과학교실) ;
  • 전상영 (전남대학교 호르몬 연구센타)
  • Published : 2006.03.31

Abstract

Objective: Pituitary adenylate cyclase-activating polypeptide (PACAP), a novel hypothalamic neuropeptide, has been suggested to play a role in ovarian folliculogenesis. The present study evaluated the effect of PACAP on the growth of preantral follicles. Methods: Preantral follicles were mechanically isolated from ovaries of 21-day-old rats and cultured in groups for 3 days in serum-free medium in the absence or presence of PACAP-38 ($10^{-6}M$). Results: Treatment with PACAP-38 resulted in an increase in follicle diameter by 75% whereas treatment with follicle stimulating hormone (FSH) increased follicle diameter by 65%. PACAP-38 treatment enhanced the granulosa cell proliferation as measured by thymidine incorporation analysis. Furthermore, the production of progesterone by cultured granulosa cells and GFSHR-17 cell line was stimulated by PACAP-38. Interestingly, PACAP enhanced FSH action on stimulation of SF-1 and aromatase gene expression. Conclusion: The present results demonstrate that PACAP stimulated preantral follicle growth by potentiating proliferation and by stimulating steroidogenesis.

목 적: 본 연구는 흰쥐 난소를 실험모델로 하여 미성숙 전동 난포의 성장에 대한 pituitary adenylate cyclase-activating polypeptide (PACAP)의 영향을 얄아보고자 하였다. 연구방법: 미성숙 전동 난포를 생후 21일된 흰쥐로부터 분리하여 PACAP을 첨가하거나 첨가하지 않은 무혈청 배양액에서 3일 동안 배양하고, 푸로게스테론 호르몬의 생성, 난포의 성장, 과립막세포의 증식 및 유전자의 동태 등을 관찰하였다. 증식의 정도는 thymidine incorporation 방법으로 검색하고 유전자의 변동은 Northern 분석을 이용하였다. 결 과: PACAP으로 처리한 군은 난포의 직경이 75% 증가한 반면 난포자극호르몬인 FSH로 처리한 군은 65% 증가하였고, PACAP 처리는 과립막 세포의 증식을 강화시켰다. FSH와 PACAP 공히 배양된 흰쥐 난포의 과립막 세포와 FSH에 반응하는 세포주인 GFSHR-17에서의 프로게스테론 생성을 촉진시켰고, PACAP이 FSH의 작용을 증진시켜 SF-1과 아로마타제 유전자 발현을 촉진시켰다. 결 론: 본 연구는 PACAP이 과립막증식과 스테로이드합성을 통하여 전동 난포의 성장을 촉진함을 시사하였고, 또한, SF-1, 아로마타제 등에 대한 FSH의 작용을 도와주는 역할을 PACAP이 담당하므로 PACAP은 초기 난포성장에 필요한 난소국소인자임을 유추할 수 있었다.

Keywords

Acknowledgement

Supported by : 전남대학교

References

  1. Gougeon A. Regulation of ovarian follicular development in primates: facts and hypotheses. Endocr Rev 1996; 17: 121-55
  2. Hirshfield AN. Development of follicles in the mammalian ovary, Int Rev Cytol 1991; 124: 43-101
  3. Li R, Phillips DM, Mather JP. Activin promotes ovarian follicle development in vitro. Endocrinol 1995; 136: 849-56
  4. Drummond AE, Dyson M, Mercer JE, Findlay JK. Differential responses of post-natal rat ovarian cells to FSH and activin. Mol Cell Endocrinol 1996; 122: 21-32
  5. Smitz J, Cortvrindt R, Hu Y, Vanderstichele H. Effects of recombinant activin A on in vitro culture of mouse preantral follicles. Mol Reprod Dev 1998; 50: 294-304
  6. Levy MJ, Hernandez ER, Adashi EY, Stillman RJ, Roberts CT Jr, Leroith D. Expression of the insulin-like growth factor (IGF)-I and II and the IGF-I and -II receptor genes during postnatal development of the rat ovary. Endocrinol 1992; 131: 1202-6
  7. Baker J, Hardy MP, Zhou J, Bondy C, Lupu F, Bellve AR. Effects of an IGFI gene null mutation on mouse reproduction. Mol Endocrinol 1996; 10: 903-18
  8. Zhou J, Kumar TR, Matzuk MM, Bondy C. Insulin-like growth factor I regulates gonadotropin responsiveness in the murine ovary. Mol Endocrinol 1997; 135: 207-21
  9. Elvin JA, Matzuk MM. Mouse models of ovarian failure. Rev Reprod 1998; 3: 183-95
  10. Kimura C, Ohkubo S, Ogi K, Hosoya M, Itoh Y, Miyata A, et al. A novel peptide which stimulates adenylate cyclase: molecular cloning and characterization of the ovine and human cDNAs. Biochem Biophy Res Commun 1990; 166: 81-9
  11. Miyata A, Arimura A, Dahl RR, Minamino N, Uehara A, Jiang L, et al. Isolation of a novel 38 residue-hypothalamic polypeptide which stimulates adenylate cyclase in pituitary cells. Biochem Biophys Res Commun 1989; 164: 567-74
  12. Keren TI, Dantes A, Sprengel R, Amsterdam A. Establishment of steroidogenic granulosa cell lines expressing follicle stimulating hormone receptors. Mol Cell Endocrinol 1993; 95: R1-R10
  13. Greenwald G, Roy SY. Follicle development and its control. In: Knobil E, Neill J (eds) The Physiology of Reproduction. Raven Press, New York 1994; 629-724
  14. Kwon HB, Schuetz AW. Role of cAMP in modulating intrafollicular progesterone levels and oocyte maturation in amphibians. Dev Biol 1986; 117: 354-64
  15. Fortune JE. Ovarian follicular growth and development in mammals. Biol Reprod 1994; 50: 225-32
  16. Gelety TJ, Magoffin DA. Ontogeny of steroidogenic enzyme gene expression in ovarian theca-interstitial cells in the rat; regulation by a paracrine theca-differentiating factor prior to achieving luteinizing hormone responsiveness. Biol Reprod 1997; 56: 938-45
  17. Kotsuji F, Tominaga T. The role of granulosa and theca cell interactions in ovarian structure and function. Mierose Res Tech 1994; 27: 97-107
  18. McGee EA, Spears N, Minami S, Hsu SY, Chun SY, Billig H, et al. Preantral ovarian follicles in serum-free culture: suppression of apoptosis after activation of the cyclic guanosine 3', 5'-monophosphate pathway and stimulation of growth and differentiation by follicle-stimulating hormone. Endocrinol 1997; 138: 2417-24
  19. Zhao J, Dorland M, Taverne MAM, van der Weijden GC, Bevers MM, van den Hurk R. In vitro culture of rat pre-antral follicles with emphasis on follicular interactions. Mol Reprod Dev 2000; 55: 65-74
  20. Tonetta SA, diZerega GS. Intragonadal regulation of follicular maturation. Endocr Rev 1989; 10: 205-29
  21. Zhong Y, Kasson BG. Pituitary adenylate cyclase-activating polypeptide stimulates steroidogenesis and adenosine 3', 5'-monophosphate accumulation in cultured rat granulosa cells. Endocrinol 1994; 135: 207-13
  22. Kumar TR, Wang Y, Lu N, Matzuk MM. Follicle stimulating hormone is required for ovarian follicle maturation but not male fertility. Nat Genet 1997; 15: 201-4
  23. Uilenbroek JT, Richards JS. Ovarian follicular development during the rat estrous cycle: gonadotropin receptors and follicular responsiveness. Biol Reprod 1979; 20: 1159-65
  24. Adashi EY, Resnick CE, Brodie AM, Svoboda ME, Van Wyk JJ. Somatomedin-C-mediated potentiation of follicle-stimulating hormone-induced aromatase activity of cultured rat granulosa cells. Endocrinology 1985; 117: 2313-20
  25. Urban RJ, Garmey JC, Shupnik MA, Veldhuis JD. Insulin-like growth factor type I increases concentrations of messenger ribonucleic acid encoding cytochrome P450 cholesterol side-chain cleavage enzyme in primary cultures of porcine granulosa cells. Endocrinol 1990; 127: 2481-8