Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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Different Developmental Competence of Porcine Oocytes Selected by Brilliant Cresyl Blue Staining and Polar Body Extrusion

Brilliant Cresyl Blue 염색방법과 극체 방출 여부에 따른 돼지 체외수정용 난포란 선별 방법이 배발달에 미치는 영향

  • Kim, Yeon-Soo (Department of Animal Resources Technology, Jinju National University) ;
  • Kim, Cheol-Wook (Department of Animal Resources Technology, Jinju National University) ;
  • Kim, In-Cheol (Swine Science Division, NLRI, RDA) ;
  • Kwack, Dae-O (Department of Science Education, Gyeongsang University) ;
  • Chung, Ki-Hwa (Department of Animal Resources Technology, Jinju National University)
  • 김연수 (진주산업대학교 동물소재공학과) ;
  • 김철욱 (진주산업대학교 동물소재공학과) ;
  • 김인철 (축산과학원 양돈과) ;
  • 곽대오 (경상대학교 과학교육과) ;
  • 정기화 (진주산업대학교 동물소재공학과)
  • Published : 2009.03.31
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Abstract

The brilliant cresyl blue (BCB) has been used to select the developmental competent oocytes in pigs, goats and cows. Growing oocytes have a higher level of active glucose-6-phosphate dehydrogenase(G6PDH) compare to mature oocytes and are rarely stained compared to mature oocytes, because G6PDH converts BCB to colorless. First polar body extrusion regard as a guideline of meoisis completion. Selection of polar body extrude oocyte is more developmental competent to blastocyst than unselected. This study was conducted to compare the BCB test to the polar body extrusion on selection of developmental competent porcine oocytes for the production of blastocyst. Cumulus-Oocytes complex were exposed to 26uM BCB stain diluted in NCSU-23 for 90 min. There was no significant difference embryo development to blastocysts between BCB treated and not treated($19.58{\pm}1.99$ vs $18.75{\pm}2.27%$), which means there was no detrimental effect of BCB exposure to oocytes. Normal fertilization is not differed among treatment groups from 70.0 to 78.4% development to blastocyst, beside polyspermy did not. To compare two different selection methods, BCB test and polar body extrusion, evaluate the developmental competent of IVP embryos. BCB+PB+(blue stained and polar body extruded, $20.71{\pm}0.45%$) and BCB-PB+(colorless and polar body extruded, $20.04{\pm}l.29%$) groups are significantly (p<0.05) higher developed than those of BCB+PB-(blue stained and no polar body, $13.24{\pm}0.73%$) and BCB-PB-(colorless and no poladbody, $7.25{\pm}0.77%$). These results showed that selection of polar body extruded oocytes method is more efficient than that of BCB test.

Keywords

References

  1. Alm H, Torner H, LOhrke B, Viergutz T, Ghoneim IM, Kanitz W (2005): Bovine blastocyst development rate in vitro is influenced by selection of oocytes by brilliant cresyl blue staining before IVM as indicator for glucose-6-phosphate dehydrogenase activity. Theriogenology 63:2194-2205 https://doi.org/10.1016/j.theriogenology.2004.09.050
  2. Anguita B, Vandaele L, Mateusen B, Maes D, Van Soom A (2007): Developmental competence of bovine oocytes is not related to apoptosis incidence in oocytes, cumulus cells and blastocysts. Theriogenology 67:537-549 https://doi.org/10.1016/j.theriogenology.2006.09.004
  3. De Loos F, van Maurik P, van Beneden T, Kruip ThAMA (1992): Structure aspects of bovine oocyte maturation in vitro. Mol Reprod Dev 31:208-214 https://doi.org/10.1002/mrd.1080310308
  4. Ebner T, Moser M, Sommergruber M, Tews G (2003): Selection based on morphological assessment of oocytes and embryos at different stages of preimplantation development : A review. Hum Reprod Update 9:251-262 https://doi.org/10.1093/humupd/dmg021
  5. Ericsson SA, Boice ML, Funahashi H, Day BN (1993): Assessment of porcine oocytes using brilliant cresyl blue. Theriogenology 39:214 https://doi.org/10.1016/0093-691X(93)90069-H
  6. Gordon I (2003): Receiving the bovine oocyte. In: Laboratory Production of Cattle Embryos (Biotechnology in Agriculture No. 27) 2nd ed. Cambridge, UK: CAB International Cambridge University Press. pp 79-111
  7. Lonergan P, Rizos D, Gutierrez-Adam A, Fair T, Boland MP (2003): Oocyte and embryo quality: effect of origin, culture conditions and gene expression patterns. Reprod Dom Anim 38:259-267 https://doi.org/10.1046/j.1439-0531.2003.00437.x
  8. Manjunatha BM, Devaraj M, Ravindra JP, Nandi S (2007): Selection of developmentally competent buffalo oocytes by brilliant cresyl blue staining before IVM. Theriogenology 68:1299-1304 https://doi.org/10.1016/j.theriogenology.2007.08.031
  9. Mangia F, Epstein CJ (1975): Biochemical studies of growing mouse oocytes: preparation of oocytes and analysis of glucose-6-phosphate dehydrogenase and lactate dehydrogenase activities. Dev Biol 45:211-220 https://doi.org/10.1016/0012-1606(75)90061-5
  10. Nicholas B, Alberio R, Fouladi-Nashta AA, Webb R (2005): Relationship between low-molecular weight insulin-like growth factor-binding proteins, caspase-3 activity and oocyte quality. Biol Reprod 72:796-804 https://doi.org/10.1095/biolreprod.104.036087
  11. Roca J, Martinez E, Vazquez JM, Lucas X (1998): Selection of immature pig oocytes for homologous in vitro penetration assays with the brilliant cresyl blue test. Reprod Fertil Develop 10:479-485 https://doi.org/10.1071/RD98060
  12. Pujol M, Lopez-Bejar M, Paramio MT (2000): Selection of immature oocytes using the brilliant cresyl blue test. Theriogenology 53:466
  13. Pujol M, Lopez-Bejar M, Meterns MJ, Rodriguez-Gonzales E, Venilla E, Paramio MT (2004): Developmental competence of heifer oocytes selected using the brilliant cresyl blue(BCB) test. Theriogenology 61:735- 744 https://doi.org/10.1016/S0093-691X(03)00250-4
  14. Rodriguez-Gonzalez E, Lopez-Bejar M, Velilla E, Paramio MT (2002): Selection of prepubertal goat oocytes using the brilliant cresyl blue test. Theriogenology 57:1397-1409 https://doi.org/10.1016/S0093-691X(02)00645-3
  15. Somfai T, Kikuchi K, Medvedev S, Onishi A, Iwamoto M, Fuchimoto D, Ozawa M, Noguchi J, Kaneko H, Ohnuma K, Sato E, Nagai T (2005): Development to the blastocyst stage of immature pig oocytes arrested before the metaphase-II stage and fertilized in vitro. Animal Reproduction Science 90:307-328 https://doi.org/10.1016/j.anireprosci.2005.03.018
  16. Wassarman M (1988): The mammalian ovum In: E. Knobil and D. Neil, Editors, The Physiology of Reproduction, vol. 1, Raven Press, NY, USA, pp 69-102
  17. 김현종, 조상래, 최창용, 최선호, 한만희, 손동수, 이승수, 상병돈, 류일선, 김인철, 김성재, 김일화, 김상근, 임경순 (2007): 돼지 난모세포의 Ethanol 처리에 의한 단위발생에 있어서 극체방출란과 분할란 선별에 따른 배 발달율 비교. 한국수정란이식학회지 22:121-126
  18. 정기화, 허태영, 곽대오, 박충생, Day BN (1999): 돼지난자의 직경이 체외성숙 및 체외발달에 미치는 영향 한국수정란이식학회지 14:17-22