한국발생생물학회지:발생과생식 (Development and Reproduction)

  • 제17권1호
  • /
  • Pages.63-72
  • /
  • 2013
  • /
  • 2465-9525(pISSN)
  • /
  • 2465-9541(eISSN)
한국발생생물학회 (The Korean Society of Developmental Biology)
권호 표지
DOI QR코드

DOI QR Code

RNA Polymerase II Inhibitor, ${\alpha}$-Amanitin, Affects Gene Expression for Gap Junctions and Metabolic Capabilities of Cumulus Cells, but Not Oocyte, during In Vitro Mouse Oocyte Maturation

  • Park, Min-Woo (Department of Biomedical Science, College of Life Science, CHA University, CHA Research Institute, Fertility Center, CHA General Hospital) ;
  • Lee, Hyun-Seo (DNA Repair Research Center, Chosun University) ;
  • Kim, Eun-Young (Department of Biomedical Science, College of Life Science, CHA University, CHA Research Institute, Fertility Center, CHA General Hospital) ;
  • Lee, Kyung-Ah (Department of Biomedical Science, College of Life Science, CHA University, CHA Research Institute, Fertility Center, CHA General Hospital)
  • 투고 : 2013.02.28
  • 심사 : 2013.03.18
  • 발행 : 2013.03.31
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

A specific inhibitor of RNA polymerase II, ${\alpha}$-amanitin is broadly used to block transcriptional activities in cells. Previous studies showed that ${\alpha}$-amanitin affects in vitro maturation of cumulus-oocyte-complex (COC). In this study, we evaluated the target of ${\alpha}$-amanitin, and whether it affects oocytes or cumulus cells (CCs), or both. We treated ${\alpha}$-amanitin with different time period during in vitro culture of denuded oocytes (DOs) or COCs in comparison, and observed the changes in morphology and maturation status. Although DOs did not show any change in morphology and maturation rates with ${\alpha}$-amanitin treatment, oocytes from COCs were arrested at metaphase I (MI) stage and CCs were more scattered than control groups. To discover causes of meiotic arrest and scattering of CCs, we focused on changes of cumulus expansion, gap junctions, and cellular metabolism which to be the important factors for the successful in vitro maturation of COCs. Expression of genes for cumulus expansion markers (Ptx3, Has2, and Tnfaip6) and gap junctional proteins (Gja1, Gja4, and Gjc1) decreased in ${\alpha}$-amanitin-treated CCs. However, these changes were not observed in oocytes. In addition, expression of genes related to metabolism (Prps1, Rpe, Rpia, Taldo1, and Tkt) decreased in ${\alpha}$-amanitin-treated CCs but not in oocytes. Therefore, we concluded that the transcriptional activities of CCs for supporting suitable transcripts, especially for its metabolic activities and formation of gap junctions among CCs as well as with oocytes, are important for oocytes maturation in COCs.

키워드

과제정보

연구 과제 주관 기관 : National Research Foundation of Korea (NRF)

참고문헌

  1. Ackert CL, Gittens JE, O'Brien MJ, Eppig JJ, Kidder GM (2001) Intercellular communication via connexin43 gap junctions is required for ovarian folliculogenesis in the mouse. Dev Biol 233:258-270. https://doi.org/10.1006/dbio.2001.0216
  2. Balasubramaniam S, Wamelink MM, Ngu LH, Talib A, Salomons GS, Jakobs C, Keng WT (2011) Novel heterozygous mutations in TALDO1 gene causing transaldolase deficiency and early infantile liver failure. J Pediat Gastr Nutr 52:113-116. https://doi.org/10.1097/MPG.0b013e3181f50388
  3. Bruzzone R, White TW, Goodenough DA (1996) The cellular Internet: on-line with connexins. Bioessays 18:709-718. https://doi.org/10.1002/bies.950180906
  4. Gershon E, Plaks V, Aharon I, Galiani D, Reizel Y, Sela- Abramovich S, Granot I, Winterhager E, Dekel N (2008) Oocyte-directed depletion of connexin43 using the Cre- LoxP system leads to subfertility in female mice. Dev Biol 313:1-12. https://doi.org/10.1016/j.ydbio.2007.08.041
  5. Kastrop PM, Hulshof SC, Bevers MM, Destree OH, Kruip TA (1991) The effects of alpha-amanitin and cycloheximide on nuclear progression, protein synthesis, and phosphorylation during bovine oocyte maturation in vitro. Mol Reprod Dev 28:249-254. https://doi.org/10.1002/mrd.1080280306
  6. Kidder GM, Mhawi AA (2002) Gap junctions and ovarian folliculogenesis. Reproduction 123:613-620. https://doi.org/10.1530/rep.0.1230613
  7. Kim HJ, Sohn KM, Shy ME, Krajewski KM, Hwang M, Park JH, Jang SY, Won HH, Choi BO, Hong SH, Kim BJ, Suh YL, Ki CS, Lee SY, Kim SH, Kim JW (2007) Mutations in PRPS1, which encodes the phosphoribosyl pyrophosphate synthetase enzyme critical for nucleotide biosynthesis, cause hereditary peripheral neuropathy with hearing loss and optic neuropathy (cmtx5). Am J Hum Genet 81:552-558. https://doi.org/10.1086/519529
  8. Kim HJ, Lee HS, Kim EY, Lee KA (2012) Downstream networking of Zap70 in meiotic cell cycle of the mouse oocytes. Dev Reprod 16:59-67.
  9. Kim Y, Kim EY, Seo YM, Yoon TK, Lee WS, Lee KA (2012) Function of the pentose phosphate pathway and its key enzyme, transketolase, in the regulation of the meiotic cell cycle in oocytes. Clin Exp Reprod Med 39:58-67. https://doi.org/10.5653/cerm.2012.39.2.58
  10. Laird DW (1996) The life cycle of a connexin: gap junction formation, removal, and degradation. J Bioenerg Biomembr 28:311-318. https://doi.org/10.1007/BF02110107
  11. Lindell TJ, Weinberg F, Morris PW, Roeder RG, Rutter WJ (1970) Specific inhibition of nuclear RNA polymerase II by alpha-amanitin. Science 170:447-449. https://doi.org/10.1126/science.170.3956.447
  12. Mehlmann LM (2005) Stops and starts in mammalian oocytes: recent advances in understanding the regulation of meiotic arrest and oocyte maturation. Reproduction 130:791-799. https://doi.org/10.1530/rep.1.00793
  13. Meinecke B, Meinecke-Tillmann S (1993) Effects of alphaamanitin on nuclear maturation of porcine oocytes in vitro. J Reprod Fertil 98:195-201. https://doi.org/10.1530/jrf.0.0980195
  14. Memili E, First NL (1998) Developmental changes in RNA polymerase II in bovine oocytes, early embryos, and effect of alpha-amanitin on embryo development. Mol Reprod Dev 51:381-389. https://doi.org/10.1002/(SICI)1098-2795(199812)51:4<381::AID-MRD4>3.0.CO;2-G
  15. Sagata N (1996) Meiotic metaphase arrest in animal oocytes: its mechanisms and biological significance. Trends Cell Biol 6:22-28. https://doi.org/10.1016/0962-8924(96)81034-8
  16. Sekeris CE, Schmid W (1972) Action of alpha-Amanitin in vivo and in vitro. FEBS Lett 27:41-45. https://doi.org/10.1016/0014-5793(72)80405-8
  17. Simon AM, Goodenough DA, Li E, Paul DL (1997) Female infertility in mice lacking connexin 37. Nature 385:525-529. https://doi.org/10.1038/385525a0
  18. Spencer N, Hopkinson DA (1980) Biochemical genetics of the pentose phosphate cycle: human ribose 5-phosphate isomerase (RPI) and ribulose 5-phosphate 3-epimerase (RPE). Ann Hum Genet 43:335-342. https://doi.org/10.1111/j.1469-1809.1980.tb01567.x
  19. Unger VM, Kumar NM, Gilula NB, Yeager M (1999) Three-dimensional structure of a recombinant gap junction membrane channel. Science 283:1176-1180. https://doi.org/10.1126/science.283.5405.1176
  20. Wiesen JF, Midgley AR, Jr (1993) Changes in expression of connexin 43 gap junction messenger ribonucleic acid and protein during ovarian follicular growth. Endocrinology 133:741-746. https://doi.org/10.1210/endo.133.2.8393773