Journal of Embryo Transfer (한국수정란이식학회지)

The Korean Society of Embryo Transfer (한국수정란이식학회)
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Respiration Rates of Individual Bovine In Vivo-Produced Embryos Measured with a Novel, Scanning Electrochemical Microscopy

Scanning Electrochemical Microscopy를 이용한 한우 체내 수정란의 호흡률 조사

  • Kim, Hyun (Animal Genetic Resources Station, National Institute of Animal Science, R.D.A.) ;
  • Bok, Nan-Hee (Animal Genetic Resources Station, National Institute of Animal Science, R.D.A.) ;
  • Kim, Sung-Woo (Animal Genetic Resources Station, National Institute of Animal Science, R.D.A.) ;
  • Do, Yoon-Jung (Animal Genetic Resources Station, National Institute of Animal Science, R.D.A.) ;
  • Kim, Min-Kyu (Department of Animal Science, Graduate School of Chungnam National University) ;
  • Cho, Sang-Rae (Subtropical Genetics Resources Station, National Institute of Animal Science, R.D.A.) ;
  • Seong, Hwan-Hoo (Animal Genetic Resources Station, National Institute of Animal Science, R.D.A.) ;
  • Kim, Dong Hun (Animal Genetic Resources Station, National Institute of Animal Science, R.D.A.) ;
  • Ko, Yeoung-Gyu (Animal Genetic Resources Station, National Institute of Animal Science, R.D.A.)
  • 김현 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 복난희 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 김성우 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 도윤정 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 김민규 (충남대학교 동물자원생명과학과) ;
  • 조상래 (농촌진흥청 국립축산과학원 난지축산시험장) ;
  • 성환후 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 김동훈 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 고응규 (농촌진흥청 국립축산과학원 가축유전자원시험장)
  • Received : 2014.01.07
  • Accepted : 2014.03.25
  • Published : 2014.03.31
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Abstract

Oxygen consumption is a useful parameter for evaluating mammalian embryo quality, since individual bovine embryos was noninvasively quantified by scanning electrochemical microscopy (SECM). Recently, several approaches have been used to measure the oxygen consumption rates of individual embryos, but relationship between oxygen consumption and pregnancy rates of Hanwoo following embryo transfer has not yet been reported. In this study, we measured to investigate the correlation between oxygen consumption rate and pregnancy rates of Hanwoo embryo using a SECM. In addition to, the expression of pluripotent gene and anti-oxidant enzyme was determined using real-time PCR by extracting RNA according to the oxygen consumption of in vivo embryo. First, we found that the oxygen consumption significantly increased in blastocyst-stage embryos (blastocyst) compared to early blastocyst stage embryos, indicating that oxygen consumption reflects the embryo quality (Grade I). Oxygen consumption of blastocyst was measured using a SECM and total cell number of in vitro blastocyst was enumerated by counting cells stained by propidium iodide. The oxygen consumption or GI blastocysts were significantly higher than those of GII blastocysts ($10.2{\times}10^{15}/mols^{-1}$ versus $6.4{\times}10^{15}/mols^{-1}$, p<0.05). Total cell numbers of in vitro blastocysts were 74.8, 90.7 and 110.2 in the oxygen consumption of below 10.0, 10.0~12.0 and over $12.0{\sim}10^{15}/mols^{-1}$, respectively. Pregnant rate in recipient cow was 0, 60 and 80% in the transplantation of embryo with the oxygen consumption of below 10.0, 10.0~12.0 and over $12.0{\times}10^{15}/mols^{-1}$, respectively. GPX1 and SOD1 were significantly increased in over -10.0 group than below 10.0 groups but in catalase gene, there was no significant difference. On the other hand, In OCT-4 and Sox2, pluripotent gene, there was a significant difference (p<0.05) between the below-10.0 ($0.98{\pm}0.1$) and over 10.0 ($1.79{\pm}0.2$). In conclusion, these results suggest that measurement of oxygen consumption maybe help increase the pregnant rate of Hanwoo embryos.

Keywords

References

  1. Abe H, Shiku H, Aoyagi S and Hoshi H. 2004. In vitro cul ture and evaluation of embryos for production of high quality bovine embryos. J. Mamm. Ova. Res. 21: 22-30. https://doi.org/10.1274/jmor.21.22
  2. Abe H, Shiku H, Yokoo M, Aoyagi S, Moriyasu S, Minamihashi A, Matsue T and Hoshi H. 2006. Evaluting the quality of individual embryos with a non-invasive and highly sensitive measurement of oxygen consumption and by scanning electrochemical microscopy. J. Reprod. Dev. 52: 55-64.
  3. Agung B, Otoi T, Abe H, Hoshi H, Murakami M, Karja NWK, Murakami MK, Wongsrikeao P, Wateri H and Suzuki T. 2005. Relationship between oxygen consumption and sex of bovine in vitro fertilizwd embryos. Reprod. Dom. Anim. 40: 51-56. https://doi.org/10.1111/j.1439-0531.2004.00554.x
  4. Boiani M, Eckardt S, Leu NA, Scholer HR and McLaughlin KJ. 2003. Pluripotency deficit in clones overcome by cloneclone aggregation: epigenetic complementation. Embo. J. 22: 5304-5312. https://doi.org/10.1093/emboj/cdg507
  5. Bortvin A, Eggan K, Skalesky H, Akutsu H, Berry DL, Yanagimachi R, Page DC and Jaenisch R. 2003. Incomplete reactivation of Oct4-related genes in mouse embryos cloned from somatic nuclei cell. 2003. Mol. Reprod. Dev. 130: 1673-1680.
  6. Caamano JN, Ryoo ZY and Youngs CR. 1998. Promotion of development of bovine embryos produced in vitro by addition of cysteine and beta-mercaptoethanol to a chemically defined culture system. J. Dairy Sci. 81: 369-374. https://doi.org/10.3168/jds.S0022-0302(98)75586-9
  7. Cho C, Son JK, Cho SR, Son JK, Choi SH, Cho CY, Kim JB, Kim SJ, Kang D and Son DS. 1998. Changs in oxygen consumption rates of embryos in Korean cattle. J. Emb. Trans. 24: 231-235.
  8. Cho C, Son JK, Cho SR, Kang D, Yeon SH, Choi SH, Choi SH, Kim NT, Kim JB, Kim SJ, Jung JW, Bok NH, Yoo YH and Son DS. 2010. Effect of pregnant rate after embryo transfer in oxygen consumpution of embryo in Korean cattle. J. Emb. Trans. 25: 145-148.
  9. Houghton FD, Thompson JG, Kennedy CJ and leese HJ. 1996. Oxygen consumption and energy metabolism of the early mouse embryo. Mol. Reprod. Dev. 44: 476-485. https://doi.org/10.1002/(SICI)1098-2795(199608)44:4<476::AID-MRD7>3.0.CO;2-I
  10. Kirchhof K, Carnwath JW, Lemme E and Anastassiadis. 2000. Expression pattern of Oct-4 in preimplantation embryos of different species. Biol. Reprod. 63: 1698-1705. https://doi.org/10.1095/biolreprod63.6.1698
  11. Kardymowicz O. 1972. A method of vital staining for determining the viability of fertilized sheep ova stored in vitro. Proc 7th Intermational Congress on Animal Reprodution & AI, Munich. 1: 503-506.
  12. Legge M and Sellens MH. 1991. Free radical scavengers ameliorate the 2-cell block in mouse embryos culture. Hum. Reprod. 5: 867-871.
  13. Lim JM and Hansel W. 1998. Improved development of in vitro-derived bovine embryos by use of a nitric oxide scanvenger in a cumulus-granulosa cell culture system. Mol. Reprod. Dev. 50: 45-53. https://doi.org/10.1002/(SICI)1098-2795(199805)50:1<45::AID-MRD6>3.0.CO;2-X
  14. Lopez AS, Larsen LH, Ramsing N, Lovendahl P, Raty M, Peippo J, Greve T and Callesen H. 2005. Respiration rates of individual bovine in vitro-produced embryos measured with a novel, non-invasive and highly sensitive microsensor system. Reproduction 130: 669-679. https://doi.org/10.1530/rep.1.00703
  15. Lopez AS, Greve T and Callesen H. 2007a. Quantification of embryo quality by respirometry. Theriogenology. 67: 21-31. https://doi.org/10.1016/j.theriogenology.2006.09.026
  16. Lopez AS, Madsen SE, Ramsing NB, Lovendahl P, Greve T and Callesen H. 2007b. Investigation of respiration of individual bovine embryos produced in vivo and in vitro and correlation with viability following transfer. Hum. Reprod. 22: 558-566. https://doi.org/10.1093/humrep/del404
  17. Magnusson C, Hillensjo T, Hamberger L and Nilsson L. 1986. Oxygen consumption by human oocytes and blastocysts grown in vitro. Hum. Reprod. 1: 183-184. https://doi.org/10.1093/oxfordjournals.humrep.a136377
  18. Niwa H. 2001. Molecular mechanism to maintain stem cell renewal of ES cells. Cell Struct. Funct. 26: 137-148. https://doi.org/10.1247/csf.26.137
  19. Nordhoff V, Hubnerm K, Bauerm A, Orlova I, Malapetsa A and Scholer HR. 2001. Comparative analysis of human, bovine, and murine Oct-4 upstream promoter sequences. Mamm. Genome. 12: 309-17. https://doi.org/10.1007/s003350010279
  20. Okamoto K, Okazawa H, Okuda A, Sakai M and Muramatsu M. 1990. A novel octamer binding transcription factor is differentially expressed in mouse embryonic cells. Cell. 60: 461-472. https://doi.org/10.1016/0092-8674(90)90597-8
  21. Overstrom EW, Burke PA, Hagopian SS and Selgraph JP. 1989. Blastocyst oxidative metabolism and embryo viability. J. Cell. Biol. 107: 607.
  22. Overstrom EW, Duby RT, Dobrinsky J, Roche JF and Boland MP. 1992. Viablity and oxydative metabolism of the bovine blastocyst. Theriogenology 37: 269. https://doi.org/10.1016/0093-691X(92)90338-R
  23. Renard JP, Philippon A and Menezo Y. 1980. In vitro uptake of glucose by bovine blastocysts. J. Reprod. Fertil. 58: 161-164. https://doi.org/10.1530/jrf.0.0580161
  24. Sakagami N, Akiyama K and Nakazawa Y. 2007. The relationship between oxygen consumption rate and pregnancy rate of bovine embryos. Reprod. Fertil. Dev. 19: 225.
  25. Sakagami N, Yamamoto Y, Akitama K, Nakazawa Y, Kojima N, Nishida K and Yoshioka K. 2010. Viability of porcine films. J. Reprod. Dev. 56: 279-284. https://doi.org/10.1262/jrd.09-101H
  26. Shiku H, Shiraishi T, Aoyagi S, Utsumi Y, Matsudaira M, Abe H, Hoshi H, Kasai S, Ohya H and Matsue T. 2004. Respiration activity of single bovine embryos entrapped in a cone-shaped microwell monitored by scanning electrochemical microscopy. Analytica Chimica Acta. 522: 51-58. https://doi.org/10.1016/j.aca.2004.06.054
  27. Shiku H, Shiraishi T, Ohya H, Matsue T, Abe H, Hoshi H and Kobayashi M. 2001. Oxygen consumption of single bovine embryos probed with scanning electrochemical microscopy. Anal. Chem. 73: 3751-3758. https://doi.org/10.1021/ac010339j
  28. Schilling EE, Goldenberg H, Morre DJ and Crane FL. 1979. Distribution of insulin receptor among mouse liver endomembranes. Biochim. Biophys. Acta. 23: 504-11.
  29. Takahashi Y and First NL. 1992. In vitro development of bovine one-cell embryo : influence of glucose, lactose, pyruvate, amino acids and vitamine. Theriogenology 37: 963- 978. https://doi.org/10.1016/0093-691X(92)90096-A
  30. Takahashi M, Nagai T, Hamano S, Kuwayama M, Okamura N and Okano A. 1993. Effect of thiol compounds on in vitro development and intracellular glutathione content of bovine embryos. Biol. Reprod. 49: 228-32. https://doi.org/10.1095/biolreprod49.2.228
  31. Trimarchi JR, Liu L, Smith PJ and Keefe DL. 2000. Noninvasive measurement of potassium efflux as an early indicator of cell death in mouse embryo. Biol. Reprod. 63: 851-857. https://doi.org/10.1095/biolreprod63.3.851
  32. Umoako Y, Noda Y, Narimoto K and Mori T. 1992. Effects of oxygen toxicity on early development of mouse embryos. Mol. Reprod. Dev. 31: 28-33. https://doi.org/10.1002/mrd.1080310106