Changes in Oxygen Consumption Rates of Embryos in Korean Cattle

한우 수정란의 발달 단계별 산소 소비량 변화

  • Choe, Chang-Yong (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Cho, Sang-Rae (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Son, Jun-Kyu (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Choi, Sun-Ho (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Cho, Chang-Yeon (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Kim, Jae-Bum (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Kim, Sung-Jae (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Kang, Da-Won (Dept. of Physiology, Institute of Health Sciences, Gyeongsang National University School of Medicine) ;
  • Son, Dong-Soo (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
  • 최창용 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 조상래 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 손준규 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 최선호 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 조창연 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 김재범 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 김성재 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 강다원 (경상대학교 의학전문대학원 생리학교실) ;
  • 손동수 (농촌진흥청 국립축산과학원 가축유전자원시험장)
  • Published : 2009.09.30

Abstract

Oxygen consumption has been regarded as a useful indicator for assessment of mammalian embryo quality. However, there was no standard criterion to measure the oxygen consumption of embryos. Here, we measured oxygen consumption of bovine embryos at various developmental stages was measured using a scanning electrochemical microscopy (SECM). We found that the oxygen consumption significantly increased in blastocyst-stage embryos compared to other stage embryos (from 2-cell-stage to morula-stage), indicating that oxygen consumption reflects the cell number ($5.2{\sim}7.6{\times}10^{14}/mol\;s^{-1}$ versus $1.2{\sim}2.4{\times}10^{14}/mol\;s^{-1}$, p<0.05). In the morula-stage embryos, the oxygen consumption of in vivo derived embryos was significantly higher than that of in vitro produced embryos ($4.0{\times}10^{14}/mol\;s^{-1}$ versus $2.4{\times}10^{14}/mol\;s^{-1}$, p<0.05). However, there was no significant difference in consumption of oxygen by in vivo and in vitro-derived bovine blastocyst-stage embryos (p>0.05). In the frozen-thawed blastocyst-stage embryos, live embryos showed significantly higher oxygen consumption than dead embryos ($4.7{\times}10^{14}/mol\;s^{-1}$ versus $1.0{\times}10^{14}/mol\;s^{-1}$, p<0.05). These results indicate that the measuring oxygen consumption by SECM can be used to evaluate bovine embryo quality.

Keywords

References

  1. Abe H, Shiku H, Aoyagi S and Hoshi H. 2004. In vitro culture 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. Evaluating the quality of individual embryos with a non-invasive and highly sensitive measurement of oxygen consumption by scanning electrochemical microscopy. J. Reprod. Dev. 52:S55-S64
  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 fertilized embryos. Reprod. Dom. Anim. 40:51-56 https://doi.org/10.1111/j.1439-0531.2004.00554.x
  4. 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
  5. Kardymowicz O. 1972. A method of vital staining for determining the viability of fertilized sheep ova stored in vitro. Proc. 7th International Congress on Animal Reproduction & AI, Munich. 1:503-506
  6. Lopez AS, Greve T and Callesen H. 2007. Quantification of embryo quality by respirometry. 2007a. Theriogenology 67:21-31 https://doi.org/10.1016/j.theriogenology.2006.09.026
  7. 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
  8. 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
  9. 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
  10. Overstrom EW, Burke PA, Hagopian SS and Selgraph JP. 1989. Blastocyst oxidative metabolism and embryo viability. J. Cell Biol. 107:607(Abstract)
  11. Overstrom EW, Duby RT, Dobrinsky J, Roche JF and Boland MP. 1992. Viability and oxidative metabolism of the bovine blastocyst. Theriogenology 37:269(Abstract) https://doi.org/10.1016/0093-691X(92)90338-R
  12. 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
  13. Schilling E, Niemann H, Cheng SP and Duepke HH. 1979. DAPI-a further fluorescence test for diagnosing the viability of early cow and rabbit ernrbyos. Zuchthyg, 14:170-172 https://doi.org/10.1111/j.1439-0531.1979.tb00936.x
  14. 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
  15. 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
  16. Thimier M. 2007. New records in the numbers of both in vivoderived and in vitro-produced bovine embryos around the world in 2006. IETS Newsletter 25:15-20
  17. Trimarchi JR, Liu L, Smith PJS and Keefe DL. 2000. Nonin-vasive measurement of potassium efflux as an early indicator of cell death in mouse embryos. Biol. Reprod. 63:851-857 https://doi.org/10.1095/biolreprod63.3.851
  18. 손동수, 최창용, 조상래, 최선호, 김현종, 김일화. 2008a. 한우 공란우의 1차 과배란처리 반응 불량개체의 2차 반응. 한국수정란이식학회지 춘계학술대회 p 75
  19. 손동수, 최창용, 조상래, 최선호, 김현종. 2008b. 2007년도 한우 및 젖소에서 수정란 생산 및 이식 실태. 제8회 발생공학 국제심포지엄 및 학술대회 p 46-47