Korean Journal of Veterinary Research (대한수의학회지)

The Korean Society of Veterinary Science (대한수의학회)
권호 표지

Calcium current on cryopreservation in mouse oocytes

동결 생쥐 난자에서의 calcium 전류

  • Kang, Da-Won (College of Medicine, Gyeongsang National University) ;
  • Kim, Eun-Sim (College of Medicine, Gyeongsang National University) ;
  • Choe, Chang-Yong (Namwon Branch Institute, National Livestock Research Institute, RDA) ;
  • Park, Jae-Yong (College of Medicine, Gyeongsang National University) ;
  • Han, Jae-Hee (College of Medicine, Gyeongsang National University) ;
  • Hong, Seong-Geun (College of Medicine, Gyeongsang National University)
  • Accepted : 2002.02.07
  • Published : 2002.03.25
Download PDF
⟨ Previous Next ⟩

Abstract

Cryopreservation is commonly used for an efficient utilization of semen, oocytes and embryos but has disadvantage in the survival, development of the post-thawed eggs. The high risk in the survival, development of eggs after thawing is thought to be caused by inappropriate internal regulation of $Ca^{2+}$ and/or formation of intracellular ice crystals. In this experiment, we tested whether the $Ca^{2+}$ current (iCa), a decisive factor to $Ca^{2+}$ entry, was altered in post-thawed oocytes by using whole cell voltage clamp technique. The quality and survival rates of the oocytes derived from both fresh and frozen groups were examined by morphology and FDA-test. Vitrified oocytes (VOs) were incubated for 4 hr after thawing and then donated to this experiment. Ethyleneglycol-ficoll-galactose (EFG) was used as a cryoprotectant for vitrification. The membrane potential was held at -80 mV and step depolarizations of 250 ms were applied from -50 mV to 50 mV in 10 mV increments. The survival rates showed a higher in VOs vitrified with EFG containing $Ca^{2+}$ than in VOs vitrified with EFG under the $Ca^{2+}$-free condition (82.0% vs 14%). In group with/without $Ca^{2+}$, the survival rates were significantly (P<0.01) difference. In the fresh metaphase II oocytes (FOs), current-voltage (I-V) relationship showed that iCa began to activate at -40 mV and reached its maximum at -10 mV. With same voltage pulses, inward currents were elicited in VOs. I-V relationships observed in VOs were similar to those in FOs. Time constants of activation and inactivation of the inward current shown in VOs were not different to those in FOs. This accordance in I-V relations and time constants in FOs with those in VOs indicates that the inward currents in FOs are unaltered by vitrification and thawing. Therefore, vitrification with EFG does not play as a factor to deteriorate $Ca^{2+}$ entry across the membrane of the oocytes.

Keywords

References

  1. Whittingham DG, Leibo SP, Mazur P. Survival of mouse embryos frozen to -$196^{\circ} C$ and -$296^{\circ} C$ . Science, 178:411, 1972
  2. Whittingham DG. Fertilization in vitro and development to term of unfertilized mouse oocytes previously stored at -$196^{\circ} C$ . J. Reprod. Fert, 49:89-94, 1977
  3. Rall WF, Fahy GM. Ice-free cryopreservation of mouse embryos at -$196^{\circ} C$ by vitrification. Nature, 313:573-575, 1985
  4. Nakagata N. Survial of 4-cell mouse embryos derived from in vitro fertilization after ultrarapid freezing and thawing. Exp. Anim, 38(3):279-282, 1989a
  5. Al-Hasani S, Dieddrich K, Van der Ven H, et al. Cryopreservation of human oocytes. Hum Reprod, 2:695-700, 1987
  6. Imoedehe DG, Sigue AB. Survival of human oocytes cryopreserved with or without the cumulus in 1,2-propanediol. J. Assist Reprod Genet, 9:323-327, 1992
  7. Carroll J, Warnes GM, Matthews CD. Increase in digyny explains polyploidy after in-vitro fertilization of frozen-thawed mouse oocytes. J. Reprod fertil, 85:489-494, 1989
  8. George MA, Johnson MH. Cytoskeletal organization and zona sensitivity to digestion by chymotrypsin of frozen-thawed mouse oocytes. Hum Reprod, 8(4):612-620, 1993
  9. Pickering SJ, Johnson MH. The influence of cooling on the meiotic spindle of the mouse oocyte. Hum Reprod, 2(3):207-216, 987
  10. Pickering SJ, Braude PR, Johnson MH, et al. Transient cooling to room temperature can cause irresversible disruption of the meiotic spindle in the human oocyte. Fertil Steril, 54:102-108, 1990
  11. Kang DW, Yi SM, Kim ES, et al. Electrical and phamacological properties of T-type calcium current in mouse oocytes. Theriogernology (Abs), 55(1):322, 2001
  12. 김양미. $Ca^2^+$ dependency of transient outward currents in hamster eggs. 석사학위논문, 1993
  13. 김양미. 햄스터 난자에 존재하는 $Cl^-$ 통로의 성상. 박사학위논문, 1997
  14. 한재희. 햄스터 난자의 수정 후 초기 분화와 관련된 전압 의존성 전류의 변화. 박사학위논문, 1995
  15. De Felici M, Siracusa G. Survival of isolated, fully grown mouse oocytes is strictly dependent on external $Ca^2^+$. Dev Biol, 92:539-543, 1982
  16. Rapael AF, James MR. Intracellular $Ca^2^+$ response of rabbit oocytes to electrical stimulation. Mol Reprod. Develop, 32:9-16, 1992
  17. Hamill OP, Marty A, Neher E, et al. Improved patchclamp techniques for high-resolution current recording from cells and cell-free membrane patches. PflugersArch, 391:85-100, 1981
  18. Vajta G, Booth PJ, Holm P, et al. Successful vitrification of early stage bovine in vitro produced embryos with the open pulled straw(OPS) method. Cryp-Letters, 18:191-195.1997
  19. Vajta G, Holm P, Kuwayama M, et al. Open pulled straw(OPS) vitrification: a new way to reduce cryoinjuries of bovine ova and embryos. Mol ReprodDev, 51:53-58, 1998
  20. Lane M, Gardner DK. Vitrification of mouse oocytes using a nylon loop. Mol Reprod Dev, 58(3):342-347, 2001
  21. Lane M, Bavister BD, Lyons EA, et al. Containerless vitrification of mammalian oocytes and embryos. Nat Biotechnol, 17:1234-1236, 1999a
  22. Lane M, Schoolcraft WB, Gardner DK. Vitrification of mouse and human blastocysts using a novel cryoloop container-less technique. Fertil Steril, 72:1073-1078, 1999b
  23. 강다원, 조성근, 한재희 등. 토끼전핵배의 동결보존 후 배발달. 한국가축번식학회지, 23(1):75-84, 1999
  24. 강다원, 최창용, 하란조 등. 토끼의 정상 및 핵이식배의 유리화 및 완만동결에 따른 융해 후 발달률. 한국수정란이식학회지, 13(1):1-9, 1998
  25. Gajata B, Smorag Z. Factors affecting the survival of one- and two-cell rabbit embryos cryopreserved by vitrification. Theriogenology, 39:499-506, 1993
  26. Shaw JM, Ward C, Trounson AO. Evaluation of propanediol, ethylene glycol, sucrose and antifreeze proteins on the survival of slow-cooled mouse pronuclearand 4-cell embryos. Hum Reprod, 10:396-402, 1995
  27. Kasai M, Hamaguchi Y, Zhu, SE, et al. T. High survial of rabbit morulae after vitrification in an ethylene glycol-based solution by a sample method. Biol.Reprod, 46:1042-1046, 1992
  28. Kasai M, Komi JH, Takakano A, et al. A simple method for mouse embryo cryopreservation in a low toxicity vitrification solution, without appreciable loss of viability. J. Reprod. Fert, 89:91-97, 1990
  29. Leibo SP, Oda K. High survival of mouse zygotes and embryos cooled rapidly or slowly in ethylene glycol plus polyvinylpyrrolidone. Cryo-Letters, 14:133-144,1993
  30. McWilliams RB, Gibbons WE, Leibo SP. Osmotic and physiological responses of mouse zygotes and human oocytes to mono-and disaccharides. Hum Reprod, 10(5):1163-1171, 1995
  31. Leibo SP, Mazur P. Methods for the preservation of mammalian embryos by freezing In Daniel JC, Jr(ed). Method for the preservation of mammalian Reproduction. Academic press, New york, NY, USA:179-201, 1978
  32. Ishida GM, Saito H, Ohta N, et al. The optimal equilibration time for mouse embryos frozen by vitrification with trehalose. Hum Reprod. 12(6) : 1259-1262. 1997
  33. Smorag Z, Heyman Y, Gardner V, et al. The effect of sucrose and trehalose on viability of one-and two-cell rabbit embryos. Theriogenology, 33(3):741-746, 1990
  34. Rayos AA, Takahashi Y, Hishinuma M, et al. Quick freezing of unfertilized mouse oocytes using ethylene glycol with sucrose or trehalose. J. Reprod Fertil,100(1):123-129, 1994
  35. Bailey JL, Buhr MM. $Ca^2^+$ regulation by cryopreserved bull spermatozoa in response to A23187. Cryobiology, 30(5):470-481, 1993
  36. Parrish JJ, Susko-Parrish JL, Graham JK. In vitro capacitation of bovine spermatozoa ; role of intracellular calcium. Theriogenology, 15 ; 51(2):461-472, 1999
  37. Szasz F, Gabor G, Solti L. Comparative study of different methods for dog semen cropreservation and testing under clinical conditions. Acta Vet Hung, 48(3):325-333. 2000
  38. Zhao Y, Buhr MM. Cryopreservation extenders affect calcium flux in bovine spermatozoa during a temperature challenge. J Androl, 16(3);278-285,1995
  39. Bae IH. Role of Calcium in resumption of meiosis of cultured porcine cumulus-enclo sed oocytes. Biol Reprod, 24:92, 1981
  40. Bae IH, Channing CP. Effect of Calcium ion the maturation of cumulus-enclosed pig follicular oocytes isolated from medium-sized Graafian follicles. Biol Reprod,79-87, 1985
  41. Homa ST, Caroll J, Swann K. The role of calcium in mammalian oocyte maturation and egg activation. Hum Reprod, 8(8):1274-1281,1993
  42. Jaffe LA. Sources of calcium in egg activation; a review and hypothesis. Dev. Biol, 99:265-276, 1983
  43. Robert MT, Calvin S, Gary GB, et al. Meiosis, egg activation, and nuclear envelope breakdown are differentially reliant on Ca2+, wherease germinal vesiclebreakdown is $Ca^2^+$ independent in the mouse oocyte. J. Cell. Biol, 117:799-811, 1992
  44. Whitaker M, Steinhardt RA. Ionic signaling in sea urchin egg fertilization. In Metz, C. & Monroy, B. (eds), Biology of fertilization, Academic Press, New York, 3:167-221, 1985
  45. Whittingham DG. Parthenogenesis in mammals. In Finn, C.A. (ed), Oxford Review in Reproduductive Biology. Vol. 2. University Press, Oxford, pp. 205-211,1980
  46. Day ML, Johnson MH, Cook DI. Cell cycle regulation of a T-type calcium in early mouse embryos. Pflugers Arch, 436(6):834-842, 1998