卵母细胞激活及其在辅助生育技术中的应用研究
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摘要
第一部分ICSI后未受精人卵母细胞的人工辅助激活研究
目的分别采用乙醇、乙醇+6-二甲基氨基嘌呤(6-DMAP)、离子霉素、离子霉素+6-DMAP、6-DMAP五种激活方法激活处理行卵胞浆内单精子注射(ICSI)后24h未受精人卵母细胞,观察和比较几种不同的人工辅助激活方法对行ICSI后24h未受精人卵母细胞的激活作用及效果。
方法收集ICSI后24h仍无受精征象的卵母细胞共计200个,分别采用上述5种方法进行激活处理。16-18h后观察第二极体(PB)排出及原核(PN)形成情况。激活后正常受精的胚胎继续培养3-5天。统计五组的激活率、原核形成情况、卵裂率及后期胚胎发育情况。采用荧光原位杂交(FISH)的方法对激活处理后正常受精发育而来的胚胎进行分析,排除孤雌激活可能。
结果五种激活方法处理后的卵母细胞激活率分别为65.6, 75.6, 83.7, 88.2和6.9%,其中离子霉素+6-DMAP组激活率最高,两种激活剂联合激活的激活率要明显高于一种激活剂单独激活(P<0.05);除离子霉素+6-DMAP组激活处理后的一原核形成能力上要明显高于6-DMAP组之外,其他组之间在各种类型原核形成能力上均无明显统计学差异(P>0.05)。FISH分析了35个从2PN发育而来的胚胎,其中有12个胚胎显示为XX信号,10个胚胎显示为XY信号。
结论通过人工辅助激活方法是可以使得ICSI后未受精的人卵母细胞恢复正常受精,并且有继续发育的潜能;离子霉素+6-DMAP组的激活率最高,但在发育的潜能上各组之间并没有统计学上的差异。
第二部分补救ICSI中人工辅助卵母细胞激活的探索研究
目的探索补救ICSI中人工辅助卵母细胞激活技术的应用,是否可以提高受精率和改善胚胎后期的发育情况,并最终可应用于临床的可行性。
方法收集常规IVF后无受精征象的人成熟卵母细胞共计148个,补救ICSI后,一组92个卵母细胞采用离子霉素+6-DMAP进行人工辅助激活处理,另一组56个未采用任何激活处理的作为对照组。16-18h后观察第二极体排出及原核形成情况。正常受精的胚胎继续培养3-5天。统计两组的受精率、原核形成情况、卵裂率及后期胚胎发育情况。采用荧光原位杂交(FISH)的方法对激活处理后2PN发育而来的胚胎进行分析,排除孤雌激活可能。
结果实验组和对照组补救ICSI后受精率分别为89.1%(82/92)和78.6%(44/56),两组之间受精率比较有显著的统计学差异(P<0.05),正常受精(2PN)率分别为51.2%(42/82)和29.5%(13/44),也有显著的统计学差异(P<0.05)。但两组之间正常受精发育的胚胎在卵裂率、优质胚胎形成率上并没有显著差异(P>0.05)。FISH分析了实验组中12个从2PN发育而来的胚胎,其中4个为XY杂交信号,5个为XX杂交信号,性染色体正常的胚胎比例为75%。
结论采用卵母细胞人工辅助激活可以提高补救ICSI的受精率,激活后受精的胚胎有进一步发育的潜能,FISH分析排除了孤雌激活,故人工辅助激活在改善补救ICSI的受精率上有一定的临床应用价值。
Part I Study on the Effect of Activation on human unfertilized oocytes after intracytoplasmic sperm injection
Objective
To investigate the effect of different activation methods on human unfertilized oocytes after intracytoplasmic sperm injection (ICSI).
Design
Oocytes allocated to this study were those that did not show any signs of normal fertilization (extrusion of the second polar body, pronuclei formation) after ICSI for 24 hours, and that were still with no signs of degeneration or fragmentation. 200 oocytes were randomly subjected to one of five activation treatment, ethanol(ET), ethanol + 6-dimethylaminopurine (6-DMAP), ionomycin ( IO ) , ionomycin + 6-dimethylaminopurine (6-DMAP), 6-dimethylaminopurine (6-DMAP). Fluorescence in situ hybridization (FISH) was employed to analyze chromosomes using X and Y dual-colored probes.
Results
Fertilization occurred in 138(69%) of 200 oocytes which were allocated to the chemical activation treatment groups. The activation rates of the oocytes in the ET, ET+6-DMAP, IO, IO+6-DMAP, 6-DMAP groups were 65.6%, 75.6%, 83.7%, 88.2% and 6.9%, respectively, showing that the activation rates of combining activation treatment groups were significantly higher than those of single activation treatment groups. Except the 1PN formation rate show a significantly difference with pronuclei formation between IO and IO+6-DMAP group, there was no other significantly differences with pronuclei formation among the groups. Sex chromosome analyses indicated 12 embryos with XX and 10 embryos with XY in 35 embryos derived from activated oocytes containing 2PN.
Conclusions
(1) It is possible to activate unfertilized oocytes after ICSI with chemicals, and the normal fertilized oocytes have further developmental potential; (2)ionomycin + 6-DMAP group showed the best activation rate among the groups, however we did not find any difference in developmental potential between the activation groups.
Part II Study on the application of Oocytes Activation in Recue intracytoplasmic sperm injection
Objective
To investigate the application effect of Oocytes Activation in Rescue ICSI.
Design
Oocytes allocated to this study were those that did not show any signs of normal fertilization (extrusion of the second polar body, pronuclei formation) by 18h after IVF, and that were still with no signs of degeneration or fragmentation. 148 oocytes were randomly subjected to the study. The experimental group was that has total 92 oocytes which received chemical activation treatment after rescue ICSI , the control group has total number of 56 oocytes, without receiving chemical activation treatment after rescue ICSI. Fluorescence in situ hybridization (FISH) was employed to analyze chromosomes using X and Y dual-colored probes on the embryos that displayed two pronuclei in the experimental group.
Results
The Fertilization rates in the experimental group and the control group were 89.1%(82/92)and 78.6%(44/56),the result showed that the fertilization rate of combining activation treatment after rescue ICSI was higher than the control group. The normal fertilization rate of the two groups were 51.2%(42/82)and 29.5%(13/44)respectively, which also had a significantly difference between the experimental group and the control group. Based on the embryos formed from normal fertilized oocytes of the two groups, no significantly difference were found in the cleavage rates and blastocyst rates. Sex chromosome analysis indicated 5 embryos with XX and 4 embryos with XY in 12 embryos derived from activated oocytes containing 2PN after rescue ICSI.
Conclusions
The combination of chemical oocytes activation treatment after rescue ICSI could effectively improve the fertilized rate after Rescue ICSI. and the normal fertilized oocytes have further developmental potential. However, we did not find any difference in developmental potential in the embryos formed from normal fertilized oocytes between two groups, showing that the oocytes activation can be applicated in unfertilized oocytes which receive rescue ICSI treatment.
目的分别采用乙醇、乙醇+6-二甲基氨基嘌呤(6-DMAP)、离子霉素、离子霉素+6-DMAP、6-DMAP五种激活方法激活处理行卵胞浆内单精子注射(ICSI)后24h未受精人卵母细胞,观察和比较几种不同的人工辅助激活方法对行ICSI后24h未受精人卵母细胞的激活作用及效果。
方法收集ICSI后24h仍无受精征象的卵母细胞共计200个,分别采用上述5种方法进行激活处理。16-18h后观察第二极体(PB)排出及原核(PN)形成情况。激活后正常受精的胚胎继续培养3-5天。统计五组的激活率、原核形成情况、卵裂率及后期胚胎发育情况。采用荧光原位杂交(FISH)的方法对激活处理后正常受精发育而来的胚胎进行分析,排除孤雌激活可能。
结果五种激活方法处理后的卵母细胞激活率分别为65.6, 75.6, 83.7, 88.2和6.9%,其中离子霉素+6-DMAP组激活率最高,两种激活剂联合激活的激活率要明显高于一种激活剂单独激活(P<0.05);除离子霉素+6-DMAP组激活处理后的一原核形成能力上要明显高于6-DMAP组之外,其他组之间在各种类型原核形成能力上均无明显统计学差异(P>0.05)。FISH分析了35个从2PN发育而来的胚胎,其中有12个胚胎显示为XX信号,10个胚胎显示为XY信号。
结论通过人工辅助激活方法是可以使得ICSI后未受精的人卵母细胞恢复正常受精,并且有继续发育的潜能;离子霉素+6-DMAP组的激活率最高,但在发育的潜能上各组之间并没有统计学上的差异。
第二部分补救ICSI中人工辅助卵母细胞激活的探索研究
目的探索补救ICSI中人工辅助卵母细胞激活技术的应用,是否可以提高受精率和改善胚胎后期的发育情况,并最终可应用于临床的可行性。
方法收集常规IVF后无受精征象的人成熟卵母细胞共计148个,补救ICSI后,一组92个卵母细胞采用离子霉素+6-DMAP进行人工辅助激活处理,另一组56个未采用任何激活处理的作为对照组。16-18h后观察第二极体排出及原核形成情况。正常受精的胚胎继续培养3-5天。统计两组的受精率、原核形成情况、卵裂率及后期胚胎发育情况。采用荧光原位杂交(FISH)的方法对激活处理后2PN发育而来的胚胎进行分析,排除孤雌激活可能。
结果实验组和对照组补救ICSI后受精率分别为89.1%(82/92)和78.6%(44/56),两组之间受精率比较有显著的统计学差异(P<0.05),正常受精(2PN)率分别为51.2%(42/82)和29.5%(13/44),也有显著的统计学差异(P<0.05)。但两组之间正常受精发育的胚胎在卵裂率、优质胚胎形成率上并没有显著差异(P>0.05)。FISH分析了实验组中12个从2PN发育而来的胚胎,其中4个为XY杂交信号,5个为XX杂交信号,性染色体正常的胚胎比例为75%。
结论采用卵母细胞人工辅助激活可以提高补救ICSI的受精率,激活后受精的胚胎有进一步发育的潜能,FISH分析排除了孤雌激活,故人工辅助激活在改善补救ICSI的受精率上有一定的临床应用价值。
Part I Study on the Effect of Activation on human unfertilized oocytes after intracytoplasmic sperm injection
Objective
To investigate the effect of different activation methods on human unfertilized oocytes after intracytoplasmic sperm injection (ICSI).
Design
Oocytes allocated to this study were those that did not show any signs of normal fertilization (extrusion of the second polar body, pronuclei formation) after ICSI for 24 hours, and that were still with no signs of degeneration or fragmentation. 200 oocytes were randomly subjected to one of five activation treatment, ethanol(ET), ethanol + 6-dimethylaminopurine (6-DMAP), ionomycin ( IO ) , ionomycin + 6-dimethylaminopurine (6-DMAP), 6-dimethylaminopurine (6-DMAP). Fluorescence in situ hybridization (FISH) was employed to analyze chromosomes using X and Y dual-colored probes.
Results
Fertilization occurred in 138(69%) of 200 oocytes which were allocated to the chemical activation treatment groups. The activation rates of the oocytes in the ET, ET+6-DMAP, IO, IO+6-DMAP, 6-DMAP groups were 65.6%, 75.6%, 83.7%, 88.2% and 6.9%, respectively, showing that the activation rates of combining activation treatment groups were significantly higher than those of single activation treatment groups. Except the 1PN formation rate show a significantly difference with pronuclei formation between IO and IO+6-DMAP group, there was no other significantly differences with pronuclei formation among the groups. Sex chromosome analyses indicated 12 embryos with XX and 10 embryos with XY in 35 embryos derived from activated oocytes containing 2PN.
Conclusions
(1) It is possible to activate unfertilized oocytes after ICSI with chemicals, and the normal fertilized oocytes have further developmental potential; (2)ionomycin + 6-DMAP group showed the best activation rate among the groups, however we did not find any difference in developmental potential between the activation groups.
Part II Study on the application of Oocytes Activation in Recue intracytoplasmic sperm injection
Objective
To investigate the application effect of Oocytes Activation in Rescue ICSI.
Design
Oocytes allocated to this study were those that did not show any signs of normal fertilization (extrusion of the second polar body, pronuclei formation) by 18h after IVF, and that were still with no signs of degeneration or fragmentation. 148 oocytes were randomly subjected to the study. The experimental group was that has total 92 oocytes which received chemical activation treatment after rescue ICSI , the control group has total number of 56 oocytes, without receiving chemical activation treatment after rescue ICSI. Fluorescence in situ hybridization (FISH) was employed to analyze chromosomes using X and Y dual-colored probes on the embryos that displayed two pronuclei in the experimental group.
Results
The Fertilization rates in the experimental group and the control group were 89.1%(82/92)and 78.6%(44/56),the result showed that the fertilization rate of combining activation treatment after rescue ICSI was higher than the control group. The normal fertilization rate of the two groups were 51.2%(42/82)and 29.5%(13/44)respectively, which also had a significantly difference between the experimental group and the control group. Based on the embryos formed from normal fertilized oocytes of the two groups, no significantly difference were found in the cleavage rates and blastocyst rates. Sex chromosome analysis indicated 5 embryos with XX and 4 embryos with XY in 12 embryos derived from activated oocytes containing 2PN after rescue ICSI.
Conclusions
The combination of chemical oocytes activation treatment after rescue ICSI could effectively improve the fertilized rate after Rescue ICSI. and the normal fertilized oocytes have further developmental potential. However, we did not find any difference in developmental potential in the embryos formed from normal fertilized oocytes between two groups, showing that the oocytes activation can be applicated in unfertilized oocytes which receive rescue ICSI treatment.
引文
[1]杨增明、孙青原、夏国良主编.生殖生物学.北京科学出版社,2005:158-177
[2] Yamano S,Nakagawa K,Nakasaka H,et al.Fertilization failure and oocyte activation[J].J Med Invest,2000,47(1):1-8.
[3]张文香,曹云霞,周平等.不同来源的精子显微受精治疗男性不育[J].安徽医科大学学报,2004,39(6):468-470.
[4] Lanzedorf SM, Slusser J, MaloneyM K, et al. A preclinical evaluation of pronuclear formation by microinjection of human spermatozoa into human oocytes[J].Fertil Steril,1988,49:835-42.
[5] Palermo G, Joris H, Devroey P, Van Steirteghem A C. Pregnancies after intracytoplasmic injection of single spermatozoon into an oocyte [J]. Lancet,1992, 340:17 - 8.
[6] Hoshi K, Yanagida K, Yazawa H, Katayose H, Sato A. Intracytoplasmic sperm injection using immobilized or motile spermatozoon[J]. Fertil Steril,1995,63:1241–5
[7] Zhang J,Wang CW, Blaszcyzk BA, Grifo JM, Ozil J, Haberman E, et al. Electrical activation and in vitro development of human oocytes that fail to fertilize after intracytoplasmic sperm injection[J]. Fertil Steril,1999,72:509–12.
[8] Nakagawa K. Yamano S. Moride N et al. Effect of activation with Ca ionophore ,A23187 and puromycin on the development of human oocytes that failed to fertilize after intracytoplasmic sperm injection[J]. Fertil Steril,2001,76:148-52.
[9] Eldar-Geva T, Brooks B, Margalioth EJ, Zylber-Haran E, Gal M, Silver SJ. Successful pregnancy and delivery after calcium ionophore oocyte activation in a normozoospermic patient with previous repeated failed fertilization after intracytoplasmic sperm injection[J]. Fertil Steril,2003,79:1656–8.
[10] Moaz M. Chemical activation of oocytes in different types of sperm abnormalities in cases of low or failed fertilization after ICSI; a prospective pilot study[J]. RBM Online,2006,13:791-794.
[11] Murase Y, Araki Y, Mizuno S, Kawaguchi C, Naito M, Yoshizawa M, et al. Pregnancy following chemical activation of oocytes in a couple with repeated failure of fertilization using ICSI: case report[J]. Hum Reprod,2004,19:1604–7.
[12] Loi P, Ledda S, Fulka JJ, Cappai P, Moor RM. Development of parthenogenetic and cloned ovine embryos: Effect of activation protocols[J]. Biol. Reprod,1998,58: 1177-1187.
[13] Grupen CG, Mau JC, Mcilfatrick SM, Maddocks S, Nottle MB. Effect of 6-dimethylaminopurine on electrically activated in vitro matured porcine oocytes[J]. Mol Reprod Dev,2002,62:387-396.
[14] Motlik J, Pavlok A, Kubelka M, Kalous J, Kalab P. Interplay between cdc2 kinase and MAP kinase pathway during maturation of mammalian oocytes[J]. Theriogenology,199,49:461-469.
[15] Tesarik J, SousaM. Comparison of Ca2 + responses in human oocytes fertilized by subzonal insemination and by intracytoplasmic sperm injection. Fertil Steril,1994,62:1197-1204
[16] Rinaudo P, Pepperell JR, Buradgunta S, et al. Dissociation between intracellular calcium elevation and development of human oocytes treated with calcium ionophore[J]. Fertil Steril,1997,68:1086-1092.
[17] Ben-Yosef D, Shalgi R Oocyte activation: lessons from human infertility[J]. Trends Mol Med,2001,7:163-169.
1. Flaherty SP, Payne D, Matthews CD. Fertilization failures and abnormal fertilization after intracytoplasmic sperm injection. Hum Reprod 1998;11:155–64.
2. Ben-Yosef D and Shalgi R. Oocyte activation: lessons from human infertility.Trends Mol Med 2001;7:163-9
3. Mahutte NG and Arici A. Failed fertilization: is it predictable? Curr Opin Obstet Gynecol 2003;15: 211–8.
4. Liu J, Nagy Z, Joris H, Tournaye H, Devroey P, Van Steirteghem A. Successful fertilization and establishment of pregnancies after intracytoplasmic sperm injection in patients with globozoospermia. Hum Reprod 1995;10:626–9.
5. Esfendiari N, Javed MH, Gotlieb L, Casper RF. Completed failed fertilization after intracytoplasmic sperm injection: analysis of 10 years’data. Int J Fertil Womens Med 2005;50:187–92.
6. Sakkas D, Urner F, Bianchi PG, Bizzaro D,Wagner I, Jaquenoud N, et al. Sperm chromatin anomalies can influence decondensation after intracytoplasmic sperm injection. Hum Reprod 1996;11:837–43.
7. Schmiady H, Tandler-Schneider A, Kentenich H. Premature chromosome condensation of the sperm nucleus after intracytoplasmic sperm injection. Hum Reprod 1996;11:2239–45.
8. Nasr-Esfahani MH, Razavi S, Mardani M, Shirazi R, Javanmardi S. The effects of failed oocyte activation and sperm protamine deficiency on fertilization post ICSI. RBM Online 2007;14:422–9.
9. Tesarik J, Sousa M. More than 90% fertilization rates after intracytoplasmic sperm injection and artitificial oocyte activation with calcium ionophore. Fertil Steril 1995;63:343-9.
10. Battaglia DE, Koehler JK, Klein NA, Tucker MJ. Failure of oocyte activation after intracytoplasmic sperm injection using round-headed sperm. Fertil Steril 1997;68: 118–22.
11. Rybouchkin AV, Van der Straeten F, Quatacker J. Fertilization and pregnancy after assisted oocyte activation and intracytoplasmic sperm injection in a case of round-headed sperm associated with deficient oocyte activation capacity. Fertil Steril 1997;68:1144–7.
12. Yanagida K, Morozumi H, Katayose S et al. Successful pregnancy after ICSI with strontium oocyte activation in low fertilization case. RBM Online 2006;13:801-6
13. Yanagida K, Katayose H, Yazawa H, Kimura Y, Sato A, Yanagimachi H, et al. Successful fertilization and pregnancy following ICSI and electrical oocyte activation. Hum Reprod 1999; 14:1307–11.
14. Zhang J,Wang CW, Blaszcyzk BA, Grifo JM, Ozil J, Haberman E, et al. Electrical activation and in vitro development of human oocytes that fail to fertilize after intracytoplasmic sperm injection. Fertil Steril 1999;72:509–12.
15. Kim ST, Cha YB, Park JM, Gye MC. Successful pregnancy and delivery from frozen-thawed embryos after intracytoplasmic sperm injection using patient with mosaic Down syndrome. Fertil Steril 2001;75:445–7.
16. Nakagawa K. Yamano S. Moride N et al. Effect of activation with Ca ionophore ,A23187 and puromycin on the development of human oocytes that failed to fertilize after intracytoplasmic sperm injection. Fertil Steril 2001;76:148-52
17. Eldar-Geva T, Brooks B, Margalioth EJ, Zylber-Haran E, Gal M, Silver SJ. Successful pregnancy and delivery after calcium ionophore oocyte activation in a normozoospermic patient with previous repeated failed fertilization after intracytoplasmic sperm injection. Fertil Steril 2003;79:1656–8.
18. Moaz M. Chemical activation of oocytes in different types of sperm abnormalities in cases of low or failed fertilization after ICSI; a prospective pilot study. RBM Online 2006;13:791-794.
19. Murase Y, Araki Y, Mizuno S, Kawaguchi C, Naito M, Yoshizawa M, et al. Pregnancy following chemical activation of oocytes in a couple with repeated failure of fertilization using ICSI: case report. Hum Reprod 2004;19:1604–7..
20. Kawahara M, Mori T, Tanaka H, Shimizu H. The suppression of fragmentation by stabilization of actin filament in porcine enucleated oocytes. Theriogenology 2002;58:1081-95
21. Wang WH, Abeydeera LR, Han YM, Prather RS, Day BN. Morphologic evaluationand actin filament distribution in porcine embryos produced in vitro and in vivo. Biol Reprod 1999;60:1020-28
22. Hao Y, Lai L, Mao J, Im GS, Bonk A, Prather RS. Apoptosis and in vitro development of preimplantation porcine embryos derived in vitro or by nuclear transfer. Biol Reprod 2003;69:502-7
23. Yamanka KI, Sugimra S, Wakai T, et al. Effect of activation treatments on actin filament distribution and in vitro development of miniature pig somatic cell nuclear transfer embryos. J. Reprod. Dev 2007;53:791-800
24. Munne S, Lee A, Rosenwaks Z, Grifo J, Cohen J: Diagnosis of major chromosome aneuploidies in human preimplantation embryos. Hum Reprod 1993; 8: 2185–2191 .
25. Miyazaki S and Ito M. Calcium signals for egg activation in mammals. J Pharmacol Sci 2006;100:545-52
26. Tesarik J and Mendoza C. In vitro fertilization by intracytoplasmic sperm injection. Bioessays 1999;21:791–801.
27. Tesarik J, Rienzi L, Ubaldi F, Mendoza C and Greco E. Use of a modified intracytoplasmic sperm injection technique to overcome spermborne oocyte activation failures. Fertil Steril 2002;78:619–624.
28. Mitalipov SM, Nusser KD, Wolf DP. Parthenogenetic activation of rhesus monkey oocytes and reconstructed embryos. Biol Reprod 2001; 65:253–9.
29. Yanahida K. Complete fertilization failure in ICSI. Hum Cell 2004;17:187–93.
30. Tejera A, Molla M, De Pablo JL. Successful pregnancy and childbirth after intracytoplasmic sperm injection with calcium ionophore oocyte activation in a globozoospermic patient. Fertil Steril 2008;90:1202e1-4
31. Hoshi K, Yanagida K, Yazawa H, Katayose H, Sato A. Intracytoplasmic sperm injection using immobilized or motile spermatozoon. Fertil Steril 1995;63:1241–5.
32. Lu Q, Zhao Y, Gao X, Li Y, Ma S, Mullen S, et al. Combination of calcium ionophore A23187 with puromycin salvages human unfertilized oocytes after ICSI. Eur J Obstet Gynecol Reprod Biol 2006;126:72–6.
33. Loi P, Ledda S, Fulka JJ, Cappai P, Moor RM. Development of parthenogenetic and cloned ovine embryos: Effect of activation protocols. Biol. Reprod 1998; 58:1177-1187.
34. Grupen CG, Mau JC, Mcilfatrick SM, Maddocks S, Nottle MB. Effect of 6-dimethylaminopurine on electrically activated in vitro matured porcine oocytes. Mol Reprod Dev 2002; 62:387-396.
35. Motlik J, Pavlok A, Kubelka M, Kalous J, Kalab P. Interplay between cdc2 kinase and MAP kinase pathway during maturation of mammalian oocytes. Theriogenology 1998; 49:461-469.
36. Kikuchi K, Naito K, Noguchi J, Shimada A, Kaneko H, Yamashita M, Aoki F, Tojo H, Toyoda Y. Maturation/M-phase promoting factor: a regulator of aging in porcine oocytes. Biol Reprod 2000; 63, 715–722.
37. Nanassy L, Lee k, Javor A, et al. Effects of activation methods and culture conditions on development of parthenogenetic porcine embryos. A Reprod Sci 2008;104:264-274
[1] Nagy, Z.P, Joris, H, Liu, J, Staessen, C, Devroey, P. and Van Steirteghem, A.C. Intracytoplasmic single sperm injection of 1 day old unfertilized human oocytes [J]. Hum. Reprod, 1993, 8, 2180-2184.
[2] Kuczynski W, Dhont M, Grygoruk C, et al. Rescue ICSI of unfertilized oocytes after IVF [J] . Hum Reprod , 2002 , 17(9) :2423-2427.
[3]冯婷,钱云,刘嘉茵,等.体外受精完全失败行卵细胞胞质内单精子注射补救的应用价值探讨[J] .中华男科学杂志, 2004 ,10 (3) :175-177 , 181.
[4] Park KS , Song HB , Chun SS. Late fertilization of unfertilized human oocytes in in vit ro fertilization and int racytoplasmic sperm injection cycles : conventional insemination versus ICSI[J] . J Assist Reprod Genet , 2000 ,17 (8) :4192424
[5] Amarin ZO , Obeidat BR , Rouzi AA , et al . Intracytoplasmic sperm injection after total conventional in-vitro fertilization failure [J] . Saudi Med J , 2005 ,26 (3) : 411-415.
[6]张文香,曹云霞,周平等.不同来源的精子显微受精治疗男性不育[J].安徽医科大学学报,2004,39(6):468-470.
[7] Kably Ambe A ,Baptista Perez Avella A ,Ruiz Anguas J ,et al . Unexpected failed fertilization in vitro:ICSI reinsemination or ICSI indication for subsequent cycles[J] . Ginecol Obstet Mex ,2002 ,70 (4) :373 - 378
[8] Lombardi E, TiveronM, Inza R, et al. Live birth and normal 1– year follow - up of a baby born after transfer of cryop reserved embryos from rescue intracytop lasmic sperm injection of 1 - day - old oocytes[J]. Fertil Steril, 2003, 80 (3) : 646 - 648.
[9] Tournaye H, et al. Intracytop lasmic sperm injection versus in vitro fertilization: a randomized controlled trial and a meta - analysis of the literature[ J ]. Fertil Steril, 2002,78 (5) : 1030 - 1037
[10] Pellestor F , Andreo B , Amal F , et al . Matemal aging and chromosomal abnomalities New data drawn from in vitro unfertilized hum an oocytes[J]. Hum Genet , 2003 ,112 (2) :195 - 203.
[11] Rinaudo P, Pepperell JR,Buradgunta S, et al1 Dissociation between intracellular calcium elevation and development of human oocytes treated with calcium ionophore1 Fertil Steril, 1997, 68: 1086-1092.
[12] Nakagawa K, Yamano S, Nakagawa H, et al1 A combination of calcium ionophore and puromycin effectively p roduces human parthenogenones with one haploid pronucleus. Zygote, 2001, 9: 832-881
[1] Voronina E, Wessel GM. The regulation of oocyte maturation [ J] .Curr Top Dev Biol, 2003, 58(2): 53-110.
[2] Tosti E, Boni R. Electrical events during gamete maturation and fertilization in animals and humans [J]. Hum Reprod Update,2004,10 (1):53一65.
[3] Tosti E. Calciumion currents mediating oocyte maturation events[ J] . Reprod Biol Endocrinol, 2006, 4(9): 26.
[4] Cuomo A, Di Cristo C, Di Cosmo A, et al. Calcium currents correlatewithoocyte maturation during the reproductive cycle in Octopus vulgaris[ J] . J Exp Zool A, 2005, 303(3): 193-202.
[5] Machaca K. Increased sensitivity and clustering of elementary Ca2+ release events during oocyte maturation[ J] . Dev Biol, 2004, 275(1):170-182
[6] Deguchi R, Morisawa M. External Ca2+ is predominantly used for cytoplasmic and nuclear Ca2+ increases in fertilized oocytes of the marine bivalve Mactra chinensis[ J] . J Cell Sci, 2003, 116(1): 367-376.
[7] .Frank-Vaillant M, Jessus C, Ozon R, at al. Two distinct mechanisms control the accumulation of cyclin B 1 and Mos in Xenopus oocytes in response in progesterone. Mol Biol Cell, 1999, 10(10): 3279-3288
[8] Levesque JT, Sirard MA. Resumption of meiosis in initiated by the accumulation of cyclin B in bovine oocytes. Biol Reprod, 1996, 55(6):1427-1436
[9] De Moor CH, Richter JD. Cytoplasmic polyadenylation elements mediate masking and unmasking of cyclin B 1 mRNA.EMBO J. 1999,18:2294-2303
[10] Fan H Y, Sun Q Y.Involvement of mitogen-activated protein kinase cascade during oocyte maturation and fertilization in mammals[J].Biol Reprod,2004,70(3):535-547
[11] Verlhac MH, Lefebvre C, Kubiak JZ, et al. Mos activates MAP kinase in mouse oocytes through two opposite pathways. EMBO J. 2000, 19(22):6065-6074
[12] Gross SD, Lewellyn AL, Maller JL. A constitutively active form of the protein kinase p90Rsk 1 is sufficient to trigger the G2/M transition in Xenopus oocytes. J Biol Chem, 2001,276(49):46099-46103
[13] Araki K, Natio K, Haraguchi S, et al.Meiotic abnormalities of cmos knockout mouse oocytes: activation after first meiosis or entrance into third meiotic metaphase. Biol Reprod, 1996,93(14):7032-7035
[14] Choi T, Fukasawa K,Zhou R, et al. The Mos/mitogen-avtivated protein kinase(MAPK) pathway regulates the size and degradation of the first polar body in maturation mouse oocytes. Proc Natl Acad Sci USA,1996,93(14):7032-7035
[15] Gordo AC, He Cl, Smith S, et al. Mitogen activated protein kinase plays a significant role in metaphase II arrest, spindle morphology and maintenance of maturation of maturation promoting factor activity in bovine oocytes. Mol Reprod Dev, 2001,59(1):106-114
[16] Downs SM, Cotton J, Hunzicker-Dum M. Protein kinase C and meiotic regulation in isolated mouse oocytes. Mol Reprod Dev ,2001,58(1):101-115
[17] Sun YQ, Xia GL, Byskov AG, at al. Protein kinase C and intracellular calcium are involved in follicle-stimulating hormone-mediated meiotic resumption of cumulus cell-enclosed porcine oocytes in hypoxanthine-supplemented medium.Mol Reprod Dev,1999,53(1):51-58
[18] Viveros MM, Hirao Y, Eppig JJ, et al. Evidence that protein kinase C(PKC) participates in the meiosis I to meiosis II transition in mouse oocytes.Dev Biol,2001,235(2):330-342
[19] Tunquist BJ,Maller JI. Under arrest: cytostatic factor(CSF)-mediated metaphase arrest in vertebrate eggs[J].Genes Dev,2003,17(6):683-710.
[20] Kure-bayashi S, Miyake M, Okada K, et al. Successful implantation of in vitro-matured electro-activated oocytes in the pig [J]. Theriogenology, 2000, 53: 1105- 1119
[21] Lee S J ,Pursel V G, Chung K S. Effects of fusion media, voltage and micromanipulation on the activation of pig oocytes and blastomere development [J]. Theriogenology, 1993,39:257
[22] Yamanka KI, Sugimra S, Wakai T, et al. Effect of activation treatments on actin filament distribution and in vitro development of miniature pig somatic cell nuclear transfer embryos. J. Reprod. Dev 2007;53:791-800
[23] Liu J L, Sung L Y,Du F, et al. Differential development of rabbit embryos derived from parthenogenesis and nuclear transfer[J]. Mol ReProd Dev, 2004, 68(1):58-64.
[24] Loi P, Ledda S, Fulka JJ, Cappai P, Moor RM. Development of parthenogeneticand cloned ovine embryos: Effect of activation protocols. Biol. Reprod 1998; 58:1177-1187
[25] Shiina Y, Kameda M, Matsuyama K, et al. Role of extracellular Ca2+ changes in fertilized and activated mouse oocytes. J Riprod Fertil, 1993, 97(1):143
[26] Wang WH, Machaty Z, Abeydeera LR, et al. Parthenogenetic activation of pig oocytes with calcium ionophore and the block to sperm penetration after activation.Biol Reprod, 1998,58(6):1357
[27] Meo SC,Leal C L, Garcia J M. Activation and early parthenogenesis of bovine oocytes treated with ethanol and strontium[J]. Anim Reprod Sci,2004,81:35-46.
[28]李光鹏,孟庆刚,魏鹏等.蛋白质合成抑制剂亚胺环己酮(CHX)对猪卵母细胞体外成熟的影响.动物学报,2001,47(6):684-690.
[29] Grupen CG, Mau JC, Mcilfatrick SM, Maddocks S, Nottle MB. Effect of 6-dimethylaminopurine on electrically activated in vitro matured porcine oocytes. Mol Reprod Dev 2002; 62:387-396
[30] Nanassy L, Lee k, Javor A, et al. Effects of activation methods and culture conditions on development of parthenogenetic porcine embryos. A Reprod Sci 2008;104:264-274
[31] Zhang J,Wang CW, Blaszcyzk BA, Grifo JM, Ozil J, Haberman E, et al. Electrical activation and in vitro development of human oocytes that fail to fertilize after intracytoplasmic sperm injection. Fertil Steril 1999;72:509–12.
[32] Lu Q, Zhao Y, Gao X, Li Y, Ma S, Mullen S, et al. Combination of calcium ionophore A23187 with puromycin salvages human unfertilized oocytes after ICSI. Eur J Obstet Gynecol Reprod Biol 2006;126:72–6.
[33] Lu Q, Chen ZJ, Gao X, et al. Influences on sex chromosomes and IGF-Ⅱexpression of oocyte activation with calcium ionophore A23187 and puromycin on human unfertilized oocytes after ICSI. Fertil Steril, 2005, 84: S418.
[34] Yanagida K, Katayose H, Yazawa H, Kimura Y, Sato A, Yanagimachi H, et al. Successful fertilization and pregnancy following ICSI and electrical oocyte activation.Hum Reprod 1999; 14:1307–11.
[35] Eldar-Geva T, Brooks B, Margalioth EJ, Zylber-Haran E, Gal M, Silver SJ. Successful pregnancy and delivery after calcium ionophore oocyte activation in a normozoospermic patient with previous repeated failed fertilization after intracytoplasmic sperm injection. Fertil Steril 2003;79:1656–8.
[36] Murase Y, Araki Y, Mizuno S, Kawaguchi C, Naito M, Yoshizawa M, et al. Pregnancy following chemical activation of oocytes in a couple with repeated failure of fertilization using ICSI: case report. Hum Reprod 2004;19:1604–7
[37] Kim ST, Cha YB, Park JM, Gye MC. Successful pregnancy and delivery from frozen-thawed embryos after intracytoplasmic sperm injection using patient with mosaic Down syndrome. Fertil Steril 2001;75:445–7.
[38] Rybouchkin AV, Van der Straeten F, Quatacker J. Fertilization and pregnancy after assisted oocyte activation and intracytoplasmic sperm injection in a case of round-headed sperm associated with deficient oocyte activation capacity. Fertil Steril 1997;68:1144–7.
[39] Cibelli JB,Cunniff K,Vrana K E.Embryonic Stem cells from parthenotes[J]. Methods Enzymol,2006,418:117-135.
[2] Yamano S,Nakagawa K,Nakasaka H,et al.Fertilization failure and oocyte activation[J].J Med Invest,2000,47(1):1-8.
[3]张文香,曹云霞,周平等.不同来源的精子显微受精治疗男性不育[J].安徽医科大学学报,2004,39(6):468-470.
[4] Lanzedorf SM, Slusser J, MaloneyM K, et al. A preclinical evaluation of pronuclear formation by microinjection of human spermatozoa into human oocytes[J].Fertil Steril,1988,49:835-42.
[5] Palermo G, Joris H, Devroey P, Van Steirteghem A C. Pregnancies after intracytoplasmic injection of single spermatozoon into an oocyte [J]. Lancet,1992, 340:17 - 8.
[6] Hoshi K, Yanagida K, Yazawa H, Katayose H, Sato A. Intracytoplasmic sperm injection using immobilized or motile spermatozoon[J]. Fertil Steril,1995,63:1241–5
[7] Zhang J,Wang CW, Blaszcyzk BA, Grifo JM, Ozil J, Haberman E, et al. Electrical activation and in vitro development of human oocytes that fail to fertilize after intracytoplasmic sperm injection[J]. Fertil Steril,1999,72:509–12.
[8] Nakagawa K. Yamano S. Moride N et al. Effect of activation with Ca ionophore ,A23187 and puromycin on the development of human oocytes that failed to fertilize after intracytoplasmic sperm injection[J]. Fertil Steril,2001,76:148-52.
[9] Eldar-Geva T, Brooks B, Margalioth EJ, Zylber-Haran E, Gal M, Silver SJ. Successful pregnancy and delivery after calcium ionophore oocyte activation in a normozoospermic patient with previous repeated failed fertilization after intracytoplasmic sperm injection[J]. Fertil Steril,2003,79:1656–8.
[10] Moaz M. Chemical activation of oocytes in different types of sperm abnormalities in cases of low or failed fertilization after ICSI; a prospective pilot study[J]. RBM Online,2006,13:791-794.
[11] Murase Y, Araki Y, Mizuno S, Kawaguchi C, Naito M, Yoshizawa M, et al. Pregnancy following chemical activation of oocytes in a couple with repeated failure of fertilization using ICSI: case report[J]. Hum Reprod,2004,19:1604–7.
[12] Loi P, Ledda S, Fulka JJ, Cappai P, Moor RM. Development of parthenogenetic and cloned ovine embryos: Effect of activation protocols[J]. Biol. Reprod,1998,58: 1177-1187.
[13] Grupen CG, Mau JC, Mcilfatrick SM, Maddocks S, Nottle MB. Effect of 6-dimethylaminopurine on electrically activated in vitro matured porcine oocytes[J]. Mol Reprod Dev,2002,62:387-396.
[14] Motlik J, Pavlok A, Kubelka M, Kalous J, Kalab P. Interplay between cdc2 kinase and MAP kinase pathway during maturation of mammalian oocytes[J]. Theriogenology,199,49:461-469.
[15] Tesarik J, SousaM. Comparison of Ca2 + responses in human oocytes fertilized by subzonal insemination and by intracytoplasmic sperm injection. Fertil Steril,1994,62:1197-1204
[16] Rinaudo P, Pepperell JR, Buradgunta S, et al. Dissociation between intracellular calcium elevation and development of human oocytes treated with calcium ionophore[J]. Fertil Steril,1997,68:1086-1092.
[17] Ben-Yosef D, Shalgi R Oocyte activation: lessons from human infertility[J]. Trends Mol Med,2001,7:163-169.
1. Flaherty SP, Payne D, Matthews CD. Fertilization failures and abnormal fertilization after intracytoplasmic sperm injection. Hum Reprod 1998;11:155–64.
2. Ben-Yosef D and Shalgi R. Oocyte activation: lessons from human infertility.Trends Mol Med 2001;7:163-9
3. Mahutte NG and Arici A. Failed fertilization: is it predictable? Curr Opin Obstet Gynecol 2003;15: 211–8.
4. Liu J, Nagy Z, Joris H, Tournaye H, Devroey P, Van Steirteghem A. Successful fertilization and establishment of pregnancies after intracytoplasmic sperm injection in patients with globozoospermia. Hum Reprod 1995;10:626–9.
5. Esfendiari N, Javed MH, Gotlieb L, Casper RF. Completed failed fertilization after intracytoplasmic sperm injection: analysis of 10 years’data. Int J Fertil Womens Med 2005;50:187–92.
6. Sakkas D, Urner F, Bianchi PG, Bizzaro D,Wagner I, Jaquenoud N, et al. Sperm chromatin anomalies can influence decondensation after intracytoplasmic sperm injection. Hum Reprod 1996;11:837–43.
7. Schmiady H, Tandler-Schneider A, Kentenich H. Premature chromosome condensation of the sperm nucleus after intracytoplasmic sperm injection. Hum Reprod 1996;11:2239–45.
8. Nasr-Esfahani MH, Razavi S, Mardani M, Shirazi R, Javanmardi S. The effects of failed oocyte activation and sperm protamine deficiency on fertilization post ICSI. RBM Online 2007;14:422–9.
9. Tesarik J, Sousa M. More than 90% fertilization rates after intracytoplasmic sperm injection and artitificial oocyte activation with calcium ionophore. Fertil Steril 1995;63:343-9.
10. Battaglia DE, Koehler JK, Klein NA, Tucker MJ. Failure of oocyte activation after intracytoplasmic sperm injection using round-headed sperm. Fertil Steril 1997;68: 118–22.
11. Rybouchkin AV, Van der Straeten F, Quatacker J. Fertilization and pregnancy after assisted oocyte activation and intracytoplasmic sperm injection in a case of round-headed sperm associated with deficient oocyte activation capacity. Fertil Steril 1997;68:1144–7.
12. Yanagida K, Morozumi H, Katayose S et al. Successful pregnancy after ICSI with strontium oocyte activation in low fertilization case. RBM Online 2006;13:801-6
13. Yanagida K, Katayose H, Yazawa H, Kimura Y, Sato A, Yanagimachi H, et al. Successful fertilization and pregnancy following ICSI and electrical oocyte activation. Hum Reprod 1999; 14:1307–11.
14. Zhang J,Wang CW, Blaszcyzk BA, Grifo JM, Ozil J, Haberman E, et al. Electrical activation and in vitro development of human oocytes that fail to fertilize after intracytoplasmic sperm injection. Fertil Steril 1999;72:509–12.
15. Kim ST, Cha YB, Park JM, Gye MC. Successful pregnancy and delivery from frozen-thawed embryos after intracytoplasmic sperm injection using patient with mosaic Down syndrome. Fertil Steril 2001;75:445–7.
16. Nakagawa K. Yamano S. Moride N et al. Effect of activation with Ca ionophore ,A23187 and puromycin on the development of human oocytes that failed to fertilize after intracytoplasmic sperm injection. Fertil Steril 2001;76:148-52
17. Eldar-Geva T, Brooks B, Margalioth EJ, Zylber-Haran E, Gal M, Silver SJ. Successful pregnancy and delivery after calcium ionophore oocyte activation in a normozoospermic patient with previous repeated failed fertilization after intracytoplasmic sperm injection. Fertil Steril 2003;79:1656–8.
18. Moaz M. Chemical activation of oocytes in different types of sperm abnormalities in cases of low or failed fertilization after ICSI; a prospective pilot study. RBM Online 2006;13:791-794.
19. Murase Y, Araki Y, Mizuno S, Kawaguchi C, Naito M, Yoshizawa M, et al. Pregnancy following chemical activation of oocytes in a couple with repeated failure of fertilization using ICSI: case report. Hum Reprod 2004;19:1604–7..
20. Kawahara M, Mori T, Tanaka H, Shimizu H. The suppression of fragmentation by stabilization of actin filament in porcine enucleated oocytes. Theriogenology 2002;58:1081-95
21. Wang WH, Abeydeera LR, Han YM, Prather RS, Day BN. Morphologic evaluationand actin filament distribution in porcine embryos produced in vitro and in vivo. Biol Reprod 1999;60:1020-28
22. Hao Y, Lai L, Mao J, Im GS, Bonk A, Prather RS. Apoptosis and in vitro development of preimplantation porcine embryos derived in vitro or by nuclear transfer. Biol Reprod 2003;69:502-7
23. Yamanka KI, Sugimra S, Wakai T, et al. Effect of activation treatments on actin filament distribution and in vitro development of miniature pig somatic cell nuclear transfer embryos. J. Reprod. Dev 2007;53:791-800
24. Munne S, Lee A, Rosenwaks Z, Grifo J, Cohen J: Diagnosis of major chromosome aneuploidies in human preimplantation embryos. Hum Reprod 1993; 8: 2185–2191 .
25. Miyazaki S and Ito M. Calcium signals for egg activation in mammals. J Pharmacol Sci 2006;100:545-52
26. Tesarik J and Mendoza C. In vitro fertilization by intracytoplasmic sperm injection. Bioessays 1999;21:791–801.
27. Tesarik J, Rienzi L, Ubaldi F, Mendoza C and Greco E. Use of a modified intracytoplasmic sperm injection technique to overcome spermborne oocyte activation failures. Fertil Steril 2002;78:619–624.
28. Mitalipov SM, Nusser KD, Wolf DP. Parthenogenetic activation of rhesus monkey oocytes and reconstructed embryos. Biol Reprod 2001; 65:253–9.
29. Yanahida K. Complete fertilization failure in ICSI. Hum Cell 2004;17:187–93.
30. Tejera A, Molla M, De Pablo JL. Successful pregnancy and childbirth after intracytoplasmic sperm injection with calcium ionophore oocyte activation in a globozoospermic patient. Fertil Steril 2008;90:1202e1-4
31. Hoshi K, Yanagida K, Yazawa H, Katayose H, Sato A. Intracytoplasmic sperm injection using immobilized or motile spermatozoon. Fertil Steril 1995;63:1241–5.
32. Lu Q, Zhao Y, Gao X, Li Y, Ma S, Mullen S, et al. Combination of calcium ionophore A23187 with puromycin salvages human unfertilized oocytes after ICSI. Eur J Obstet Gynecol Reprod Biol 2006;126:72–6.
33. Loi P, Ledda S, Fulka JJ, Cappai P, Moor RM. Development of parthenogenetic and cloned ovine embryos: Effect of activation protocols. Biol. Reprod 1998; 58:1177-1187.
34. Grupen CG, Mau JC, Mcilfatrick SM, Maddocks S, Nottle MB. Effect of 6-dimethylaminopurine on electrically activated in vitro matured porcine oocytes. Mol Reprod Dev 2002; 62:387-396.
35. Motlik J, Pavlok A, Kubelka M, Kalous J, Kalab P. Interplay between cdc2 kinase and MAP kinase pathway during maturation of mammalian oocytes. Theriogenology 1998; 49:461-469.
36. Kikuchi K, Naito K, Noguchi J, Shimada A, Kaneko H, Yamashita M, Aoki F, Tojo H, Toyoda Y. Maturation/M-phase promoting factor: a regulator of aging in porcine oocytes. Biol Reprod 2000; 63, 715–722.
37. Nanassy L, Lee k, Javor A, et al. Effects of activation methods and culture conditions on development of parthenogenetic porcine embryos. A Reprod Sci 2008;104:264-274
[1] Nagy, Z.P, Joris, H, Liu, J, Staessen, C, Devroey, P. and Van Steirteghem, A.C. Intracytoplasmic single sperm injection of 1 day old unfertilized human oocytes [J]. Hum. Reprod, 1993, 8, 2180-2184.
[2] Kuczynski W, Dhont M, Grygoruk C, et al. Rescue ICSI of unfertilized oocytes after IVF [J] . Hum Reprod , 2002 , 17(9) :2423-2427.
[3]冯婷,钱云,刘嘉茵,等.体外受精完全失败行卵细胞胞质内单精子注射补救的应用价值探讨[J] .中华男科学杂志, 2004 ,10 (3) :175-177 , 181.
[4] Park KS , Song HB , Chun SS. Late fertilization of unfertilized human oocytes in in vit ro fertilization and int racytoplasmic sperm injection cycles : conventional insemination versus ICSI[J] . J Assist Reprod Genet , 2000 ,17 (8) :4192424
[5] Amarin ZO , Obeidat BR , Rouzi AA , et al . Intracytoplasmic sperm injection after total conventional in-vitro fertilization failure [J] . Saudi Med J , 2005 ,26 (3) : 411-415.
[6]张文香,曹云霞,周平等.不同来源的精子显微受精治疗男性不育[J].安徽医科大学学报,2004,39(6):468-470.
[7] Kably Ambe A ,Baptista Perez Avella A ,Ruiz Anguas J ,et al . Unexpected failed fertilization in vitro:ICSI reinsemination or ICSI indication for subsequent cycles[J] . Ginecol Obstet Mex ,2002 ,70 (4) :373 - 378
[8] Lombardi E, TiveronM, Inza R, et al. Live birth and normal 1– year follow - up of a baby born after transfer of cryop reserved embryos from rescue intracytop lasmic sperm injection of 1 - day - old oocytes[J]. Fertil Steril, 2003, 80 (3) : 646 - 648.
[9] Tournaye H, et al. Intracytop lasmic sperm injection versus in vitro fertilization: a randomized controlled trial and a meta - analysis of the literature[ J ]. Fertil Steril, 2002,78 (5) : 1030 - 1037
[10] Pellestor F , Andreo B , Amal F , et al . Matemal aging and chromosomal abnomalities New data drawn from in vitro unfertilized hum an oocytes[J]. Hum Genet , 2003 ,112 (2) :195 - 203.
[11] Rinaudo P, Pepperell JR,Buradgunta S, et al1 Dissociation between intracellular calcium elevation and development of human oocytes treated with calcium ionophore1 Fertil Steril, 1997, 68: 1086-1092.
[12] Nakagawa K, Yamano S, Nakagawa H, et al1 A combination of calcium ionophore and puromycin effectively p roduces human parthenogenones with one haploid pronucleus. Zygote, 2001, 9: 832-881
[1] Voronina E, Wessel GM. The regulation of oocyte maturation [ J] .Curr Top Dev Biol, 2003, 58(2): 53-110.
[2] Tosti E, Boni R. Electrical events during gamete maturation and fertilization in animals and humans [J]. Hum Reprod Update,2004,10 (1):53一65.
[3] Tosti E. Calciumion currents mediating oocyte maturation events[ J] . Reprod Biol Endocrinol, 2006, 4(9): 26.
[4] Cuomo A, Di Cristo C, Di Cosmo A, et al. Calcium currents correlatewithoocyte maturation during the reproductive cycle in Octopus vulgaris[ J] . J Exp Zool A, 2005, 303(3): 193-202.
[5] Machaca K. Increased sensitivity and clustering of elementary Ca2+ release events during oocyte maturation[ J] . Dev Biol, 2004, 275(1):170-182
[6] Deguchi R, Morisawa M. External Ca2+ is predominantly used for cytoplasmic and nuclear Ca2+ increases in fertilized oocytes of the marine bivalve Mactra chinensis[ J] . J Cell Sci, 2003, 116(1): 367-376.
[7] .Frank-Vaillant M, Jessus C, Ozon R, at al. Two distinct mechanisms control the accumulation of cyclin B 1 and Mos in Xenopus oocytes in response in progesterone. Mol Biol Cell, 1999, 10(10): 3279-3288
[8] Levesque JT, Sirard MA. Resumption of meiosis in initiated by the accumulation of cyclin B in bovine oocytes. Biol Reprod, 1996, 55(6):1427-1436
[9] De Moor CH, Richter JD. Cytoplasmic polyadenylation elements mediate masking and unmasking of cyclin B 1 mRNA.EMBO J. 1999,18:2294-2303
[10] Fan H Y, Sun Q Y.Involvement of mitogen-activated protein kinase cascade during oocyte maturation and fertilization in mammals[J].Biol Reprod,2004,70(3):535-547
[11] Verlhac MH, Lefebvre C, Kubiak JZ, et al. Mos activates MAP kinase in mouse oocytes through two opposite pathways. EMBO J. 2000, 19(22):6065-6074
[12] Gross SD, Lewellyn AL, Maller JL. A constitutively active form of the protein kinase p90Rsk 1 is sufficient to trigger the G2/M transition in Xenopus oocytes. J Biol Chem, 2001,276(49):46099-46103
[13] Araki K, Natio K, Haraguchi S, et al.Meiotic abnormalities of cmos knockout mouse oocytes: activation after first meiosis or entrance into third meiotic metaphase. Biol Reprod, 1996,93(14):7032-7035
[14] Choi T, Fukasawa K,Zhou R, et al. The Mos/mitogen-avtivated protein kinase(MAPK) pathway regulates the size and degradation of the first polar body in maturation mouse oocytes. Proc Natl Acad Sci USA,1996,93(14):7032-7035
[15] Gordo AC, He Cl, Smith S, et al. Mitogen activated protein kinase plays a significant role in metaphase II arrest, spindle morphology and maintenance of maturation of maturation promoting factor activity in bovine oocytes. Mol Reprod Dev, 2001,59(1):106-114
[16] Downs SM, Cotton J, Hunzicker-Dum M. Protein kinase C and meiotic regulation in isolated mouse oocytes. Mol Reprod Dev ,2001,58(1):101-115
[17] Sun YQ, Xia GL, Byskov AG, at al. Protein kinase C and intracellular calcium are involved in follicle-stimulating hormone-mediated meiotic resumption of cumulus cell-enclosed porcine oocytes in hypoxanthine-supplemented medium.Mol Reprod Dev,1999,53(1):51-58
[18] Viveros MM, Hirao Y, Eppig JJ, et al. Evidence that protein kinase C(PKC) participates in the meiosis I to meiosis II transition in mouse oocytes.Dev Biol,2001,235(2):330-342
[19] Tunquist BJ,Maller JI. Under arrest: cytostatic factor(CSF)-mediated metaphase arrest in vertebrate eggs[J].Genes Dev,2003,17(6):683-710.
[20] Kure-bayashi S, Miyake M, Okada K, et al. Successful implantation of in vitro-matured electro-activated oocytes in the pig [J]. Theriogenology, 2000, 53: 1105- 1119
[21] Lee S J ,Pursel V G, Chung K S. Effects of fusion media, voltage and micromanipulation on the activation of pig oocytes and blastomere development [J]. Theriogenology, 1993,39:257
[22] Yamanka KI, Sugimra S, Wakai T, et al. Effect of activation treatments on actin filament distribution and in vitro development of miniature pig somatic cell nuclear transfer embryos. J. Reprod. Dev 2007;53:791-800
[23] Liu J L, Sung L Y,Du F, et al. Differential development of rabbit embryos derived from parthenogenesis and nuclear transfer[J]. Mol ReProd Dev, 2004, 68(1):58-64.
[24] Loi P, Ledda S, Fulka JJ, Cappai P, Moor RM. Development of parthenogeneticand cloned ovine embryos: Effect of activation protocols. Biol. Reprod 1998; 58:1177-1187
[25] Shiina Y, Kameda M, Matsuyama K, et al. Role of extracellular Ca2+ changes in fertilized and activated mouse oocytes. J Riprod Fertil, 1993, 97(1):143
[26] Wang WH, Machaty Z, Abeydeera LR, et al. Parthenogenetic activation of pig oocytes with calcium ionophore and the block to sperm penetration after activation.Biol Reprod, 1998,58(6):1357
[27] Meo SC,Leal C L, Garcia J M. Activation and early parthenogenesis of bovine oocytes treated with ethanol and strontium[J]. Anim Reprod Sci,2004,81:35-46.
[28]李光鹏,孟庆刚,魏鹏等.蛋白质合成抑制剂亚胺环己酮(CHX)对猪卵母细胞体外成熟的影响.动物学报,2001,47(6):684-690.
[29] Grupen CG, Mau JC, Mcilfatrick SM, Maddocks S, Nottle MB. Effect of 6-dimethylaminopurine on electrically activated in vitro matured porcine oocytes. Mol Reprod Dev 2002; 62:387-396
[30] Nanassy L, Lee k, Javor A, et al. Effects of activation methods and culture conditions on development of parthenogenetic porcine embryos. A Reprod Sci 2008;104:264-274
[31] Zhang J,Wang CW, Blaszcyzk BA, Grifo JM, Ozil J, Haberman E, et al. Electrical activation and in vitro development of human oocytes that fail to fertilize after intracytoplasmic sperm injection. Fertil Steril 1999;72:509–12.
[32] Lu Q, Zhao Y, Gao X, Li Y, Ma S, Mullen S, et al. Combination of calcium ionophore A23187 with puromycin salvages human unfertilized oocytes after ICSI. Eur J Obstet Gynecol Reprod Biol 2006;126:72–6.
[33] Lu Q, Chen ZJ, Gao X, et al. Influences on sex chromosomes and IGF-Ⅱexpression of oocyte activation with calcium ionophore A23187 and puromycin on human unfertilized oocytes after ICSI. Fertil Steril, 2005, 84: S418.
[34] Yanagida K, Katayose H, Yazawa H, Kimura Y, Sato A, Yanagimachi H, et al. Successful fertilization and pregnancy following ICSI and electrical oocyte activation.Hum Reprod 1999; 14:1307–11.
[35] Eldar-Geva T, Brooks B, Margalioth EJ, Zylber-Haran E, Gal M, Silver SJ. Successful pregnancy and delivery after calcium ionophore oocyte activation in a normozoospermic patient with previous repeated failed fertilization after intracytoplasmic sperm injection. Fertil Steril 2003;79:1656–8.
[36] Murase Y, Araki Y, Mizuno S, Kawaguchi C, Naito M, Yoshizawa M, et al. Pregnancy following chemical activation of oocytes in a couple with repeated failure of fertilization using ICSI: case report. Hum Reprod 2004;19:1604–7
[37] Kim ST, Cha YB, Park JM, Gye MC. Successful pregnancy and delivery from frozen-thawed embryos after intracytoplasmic sperm injection using patient with mosaic Down syndrome. Fertil Steril 2001;75:445–7.
[38] Rybouchkin AV, Van der Straeten F, Quatacker J. Fertilization and pregnancy after assisted oocyte activation and intracytoplasmic sperm injection in a case of round-headed sperm associated with deficient oocyte activation capacity. Fertil Steril 1997;68:1144–7.
[39] Cibelli JB,Cunniff K,Vrana K E.Embryonic Stem cells from parthenotes[J]. Methods Enzymol,2006,418:117-135.