Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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Variation of Transcribed X-linked Genes in Bovine Embryos Cloned with Fibroblasts at Different Age and Cell Cycle

  • Jeon, Byeong-Gyun (OBS/Theriogeno!ogy and Biotechnology, College of Veterinary Medicine, Gyeongsang National University) ;
  • Rho, Gyu-Jin (OBS/Theriogeno!ogy and Biotechnology, College of Veterinary Medicine, Gyeongsang National University)
  • Received : 2011.05.14
  • Accepted : 2011.06.13
  • Published : 2011.06.30
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Abstract

The present study compared the developmental potential, telomerase activity and transcript levels of X-linked genes (ANT3, HPRT, MeCP2, RPS4X, XIAP, XIST and ZFX) in the bovine somatic cell nuclear transfer (SCNT) embryos derived from different age and cell cycle of female donor nucleus. In experiment 1, the fusion rate, cleavage rate to 2-cell stage, developmental rate to blastocyst stage, and the mean number of total and ICM cells was slightly increased in embryos cloned with fetal fibroblasts compared to those with adult fibroblasts, but there was no significantly (p<0.05) differences. Telomerase activity was also similar in blastocysts cloned with fetal and adult fibroblasts. Up-regulated RPS4X and down-regulated MeCP2, XIAP, and XIST transcript level were observed in blastocysts cloned with adult fibroblasts, compared to those with fetal fibroblasts. In experiment 2, the fusion rate, cleavage rate to 2-cell stage, developmental rate to blastocyst stage, and the mean number of total and ICM cells was significantly (p<0.05) increased in embryos cloned with fetal fibroblasts at early G1 phase of the cell cycle, compared to those of fetal fibroblasts at late G1 phase. DNMT1 transcript was observed to significantly (p<0.05) increased in the fetal fibroblasts at 3 hrs after trypsin treatment of confluent culture. Further, level of telomerase activity and transcribed X-linked genes was also significantly (p<0.05) higher in the early G1 SCNT blastocysts than those of late G1. The results imply that fetal fibroblasts at early G1 phase induces the enhanced developmental potential and up-regulated telomerase activity and X-linked gene, but aberrant transcript pattern of X-linked genes may be displayed in the SCNT embryos.

Keywords

References

  1. Betts D, Bordignon V, Hill J, Winger Q, Westhusin M, Smith L, King W (2001): Reprogramming of telomerase activity and rebuilding of telomere length in cloned cattle. Proc Natl Acad Sci USA 98:1077-1082.
  2. Bogdanovic O, Veenstra GJ (2009): DNA methylation and methyl-CpG binding proteins: developmental requirements and function. Chromosoma 118(5):549-565. Review. https://doi.org/10.1007/s00412-009-0221-9
  3. Boggs BA, Chinault AC (1994): Analysis of replication timing propellies of human X-chromosomal loci by fluorescence in situ hybridization. Proc Natl Acad Sci USA 91:6083-6087. https://doi.org/10.1073/pnas.91.13.6083
  4. Brown CJ, Chow JC (2003): Beyond sense: the role of antisense RNA in controlling Xist expression. Semin Cell Dev Biol 14:341-347. https://doi.org/10.1016/j.semcdb.2003.09.013
  5. Campbell KH, Loi P, Cappai P, Wilmut I (1994): Improved development to blastocyst of ovine nuclear transfer embryos reconstructed during the presumptive S-phase of enucleated activated oocytes. Biol Reprod 50(6):1385-1393. https://doi.org/10.1095/biolreprod50.6.1385
  6. Chang SC, Tucker T, Thorogood NP, Brown CJ (2006): Mechanisms of X-chromosome inactivation. Front Bio-sci 11:852-866. https://doi.org/10.2741/1842
  7. Collas P, Pinto-Correia C, Ponce de Leon FA, Robl JM (1992): Effect of donor cell cycle stage on chromatin and spindle morphology in nuclear transplant rabbit embryos. Biol Reprod 46(3):501-511. https://doi.org/10.1095/biolreprod46.3.501
  8. Guilleret I, Benhattar J (2003): Demethylation of the human telomerase catalytic subunit (hTERT) gene promoter reduced hTERT expression and telomerase activity and shortened telomeres. Exp Cell Res 289:326-334. https://doi.org/10.1016/S0014-4827(03)00281-7
  9. Harada H, Grant S (2003): Apoptosis regulators. Rev Clin Exp Hematol 7:117-138.
  10. Heard E (2004): Recent advances in X-chromosome in-activation. Current Opinion in Cell Biology 16:247-255. https://doi.org/10.1016/j.ceb.2004.03.005
  11. Hoare SF, Bryce LA, Wisman GB, Burns S, Going JJ, van der Zee AG, Keith WN (2001): Lack of telomerase RNA gene hTERC expression in alternative lengthening of telomeres cells is associated with methylation of the hTERC promoter. Cancer Res 61:27-32.
  12. Jeon BG, Coppola G, Perrault SD, Rho GJ, Betts DH, King WA (2008): S-adenosylhomocysteine treatment of adult female fibroblasts alters X-chromosome inactivation and improves in vitro embryo development after somatic cell nuclear transfer. Reproduction 135:815-828. https://doi.org/10.1530/REP-07-0442
  13. Kasinathan P, Knott JG, Moreira PN, Burnside AS, Jerry DJ, Robl JM (2001): Effect of fibroblast donor cell age and cell cycle on development of bovine nuclear transfer embryos in vitro. Biol Reprod 64(5): 1487-1493. https://doi.org/10.1095/biolreprod64.5.1487
  14. Kawahara M, Wakai T, Yamanaka K, Kobayashi J, Sugimura S, Shimizu T, Matsumoto H, Kim JH, Sasada H, Sato E (2005): Caffeine promotes premature chromosome condensation formation and in vitro development in porcine reconstructed embryos via a high level of maturation promoting factor activity during nuclear transfer. Reproduction 130:351-357. https://doi.org/10.1530/rep.1.00644
  15. Keefer CL (2008): Lessons learned from nuclear transfer (cloning). Theriogenology 69:48-54. https://doi.org/10.1016/j.theriogenology.2007.08.033
  16. Kimura F, Seifert HH, Florl AR, Santourlidis S, Stein-hoff C, Swiatkowski S, Mahotka C, Gerharz CD, Schulz WA (2003): Decrease of DNA methyltransferase 1 expression relative to cell proliferation in transitional cell carcinoma. Int J Cancer 1;104(5):568-578. https://doi.org/10.1002/ijc.10988
  17. Lin MJ, Lee TL, Hsu DW, Shen CK (2000): One-codon alternative splicing of the CpG MTase Dnmtl transcript in mouse somatic cells. FEBS Lett 3;469(1):101-104. https://doi.org/10.1016/S0014-5793(00)01254-0
  18. Liu JH, Yin S, Xiong B, Hou Y, Chen DY, Sun QY (2008): Aberrant DNA methylation imprints in aborted bovine clones. Mol Reprod Dev 75:598-607. https://doi.org/10.1002/mrd.20803
  19. Miyamoto K, Hoshino Y, Minami N, Yamada M, Imai H (2007): Effects of synchronization of donor cell cycle on embryonic development and DNA synthesis in porcine nuclear transfer embryos. J Reprod Dev 53(2):237-246. https://doi.org/10.1262/jrd.18085
  20. Niemann H, Tian XC, King WA, Lee RS (2008): Epigenetic reprogramming in embryonic and foetal development upon somatic cell nuclear transfer cloning. Reproduction 135:151-163. https://doi.org/10.1530/REP-07-0397
  21. Nino-Soto MI, Basrur PK, King WA (2007): Impact of in vitro production techniques on the expression of X-linked genes in bovine (Bos taurus) oocytes and pre-attachment embryos. Mol Reprod Dev 74(2):144-153. https://doi.org/10.1002/mrd.20575
  22. Nolen LD, Gao S, Han Z, Mann MR, Gie Chung Y, Otte AP, Bartolomei MS, Latham KE (2005): X chromosome reactivation and regulation in cloned embryos. Dev Biol 279:525-540. https://doi.org/10.1016/j.ydbio.2005.01.016
  23. Oback B (2008): Climbing mount efficiency--small steps, not giant leaps towards higher cloning success in farm animals. Reprod Domest Anim 43 Suppl 2:407-416. https://doi.org/10.1111/j.1439-0531.2008.01192.x
  24. Santos P, Chaveiro A, Simoes N, Moreira da Silva F (2008): Bovine oocyte quality in relation to ultrastructural characteristics of zona pellucida, polyspermic penetration and developmental competence. Reprod Domest Anim 43(6):685-689. https://doi.org/10.1111/j.1439-0531.2007.00970.x
  25. Shin MR, Park SW, Shim H, Kim NH (2002): Nuclear and microtubule reorganization in nuclear-transferred bovine embryos. Mol Reprod Dev 62:74-82. https://doi.org/10.1002/mrd.10069
  26. Thouas GA, Korfiatis NA, French AJ, Jones GM, Trounson AO (2001): Simplified technique for differential staining of inner cell mass and trophectoderm cells of mouse and bovine blastocysts. Reprod Biomed Online 3:25-29. https://doi.org/10.1016/S1472-6483(10)61960-8
  27. Yang C, Przyborski S, Cooke MJ, Zhang X, Stewart R, Anyfantis G, Atkinson SP, Saretzki G, Armstrong L, Lako M (2008): A key role for telomerase reverse transcriptase unit in modulating human embryonic stem cell proliferation, cell cycle dynamics, and in vitro differentiation. Stem Cells 26:850-863. https://doi.org/10.1634/stemcells.2007-0677
  28. Zamora M, Granell M, Mampel T, Vinas O (2004): Adenine nucleotide translocase 3 (ANT3) overexpression induces apoptosis in cultured cells. FEBS Lett 563:155-160. https://doi.org/10.1016/S0014-5793(04)00293-5