线粒体移植对水牛卵母细胞发育潜能的影响

doi:10.11843/j.issn.0366-6964.2015.04.011

畜牧兽医学报 ›› 2015, Vol. 46 ›› Issue (4): 583-591.doi: 10.11843/j.issn.0366-6964.2015.04.011

线粒体移植对水牛卵母细胞发育潜能的影响

高亚可，陆凤花，吴柱连，马帆，刘晓华，杜姗姗，石德顺^*

（广西大学亚热带生物资源保护利用国家重点实验室，南宁 530005）

收稿日期:2014-10-30 出版日期:2015-04-23 发布日期:2015-04-23
通讯作者: 石德顺，博导，研究员，主要从事动物胚胎工程与转基因动物研究，E-mail：ardsshi@gxu.edu.cn
作者简介:高亚可（1987-），男，河南许昌人，硕士生，主要从事动物胚胎工程研究，E-mail：gaoyake_2008@163.com
基金资助:
国家“863”项目（2011AA100607）；教育部高等学校博士学科点博导基金项目（20114501110001）；教育部回国基金项目（AC150070）；广西科学基金项目（2012GXNSFFA060004）；广西研究生教育创新计划资助项目（T3340098603）

Effects of Mitochondrial Transplant on the Developmental Potential of Buffalo Oocytes

GAO Ya-ke，LU Feng-hua，WU Zhu-lian，MA Fan，LIU Xiao-hua，DU Shan-shan，SHI De-shun^*

(State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources，Guangxi University，Nanning 530005，China)

Received:2014-10-30 Online:2015-04-23 Published:2015-04-23

摘要/Abstract

摘要：

本研究以水牛卵泡颗粒细胞作为线粒体的来源细胞，初步探讨水牛卵母细胞进行线粒体移植（MIT）后对其发育潜能的影响。试验比较了不同级别水牛卵母细胞mtDNA的拷贝数，并研究了水牛卵母细胞进行MIT后，其后续早期胚胎发育及胚胎线粒体膜电位（ΔΨm）的变化情况。结果显示：一级卵母细胞组的平均mtDNA拷贝数极显著高于二、三级卵母细胞组（（202 101±74 432） vs （118 483±17 028），（39 177±7 938），P＜0.01），二级卵母细胞组的平均mtDNA拷贝数极显著高于三级卵母细胞组（（118 483±17 028） vs （39 177±7 938），P＜0.01）；孤雌激活处理后发现：一级卵母细胞组的卵裂率和囊胚率也都极显著高于二、三级卵母细胞组（P＜0.01），而二级卵母细胞组激活后胚胎的卵裂率和囊胚率亦极显著高于三级卵母细胞组（P＜0.01）。用Mito-Tracker探针标记的外源线粒体经移植后，会随着胚胎的发育而发生移动，分布到各个卵裂球中；且发现二级卵母细胞MIT组孤雌激活后的囊胚率显著高于对照组和空注组（27.3% vs 17.4%，7.84%，P＜0.05），而三级卵母细胞MIT组激活后的囊胚率与对照组和空注组差异不显著（P>0.05）；对胚胎ΔΨm检测发现，水牛植入前各时期胚胎ΔΨm总体呈上升趋势，线粒体移植后胚胎各时期的ΔΨm均显著高于对照组（P＜0.05）。综上表明：不同质量的水牛卵母细胞其mtDNA拷贝数存在显著差异，且mtDNA拷贝数与卵母细胞质量和发育能力成正相关；通过移植外源线粒体可以提高二级水牛卵母细胞的发育潜能。

Abstract:

The aim of present study was to investigate the effects of mitochondrial transplant (MIT) on the developmental potential of buffalo oocytes.The mitochondrial DNA (mtDNA) copy number in the oocyte，and the developmental competence，exogenous mitochondria distribution，as well as mitochondrial membrane potential (ΔΨm) in the preimplantation embryos after the MIT were examined.The result showed that the average mtDNA copy number of buffalo oocytes in the first level of oocytes was evidently higher than the second and the third level of oocytes (（202 101±74 432） vs （118 483±17 028），（39 177±7 938），P＜0.01），and the average mtDNA copy number in the second level of oocytes was significantly higher than that of the third level of oocytes (（118 483±17 028） vs （39 177±7 938），P<0.01).Furthermore，the cleavage rates and blastocyst rates of the first level of oocytes were significantly higher than those of the second and the third level of oocytes after parthenogenetic activation (P<0.01)，and the cleavage rates and blastocyst rates of the second level of oocytes were remarkably higher than those of the third level of oocytes (P<0.01).The exogenetic mitochondria labeled with Mito-Tracker fluorescent probe distributed into each of blastomeres with the embryonic development after MIT.Moreover，more the second level of oocytes that received MIT developed to the blastocyst stage after parthenogenetic activation in comparison with oocytes that did not received MIT or were injected with solution (27.3% vs 17.4%，7.84%，P<0.05)，however，there were no significant difference in the blastocyst rates of the third level of oocytes than the control group and the nothing injection group after MIT.The ΔΨm of buffalo preimplantation embryo presented an increased trend during embryonic development，and the ΔΨm of embryo derived from MIT group was significantly higher than that of the control group (P<0.05).These results showed that the mtDNA copy numbers in different quality buffalo oocytes have significant differences，and the mtDNA copy numbers of buffalo oocytes are positively related with the quality of oocytes and their developmental potential，and the developmental ability of the second level of oocytes can be improve by MIT.

中图分类号:

高亚可，陆凤花，吴柱连，马帆，刘晓华，杜姗姗，石德顺. 线粒体移植对水牛卵母细胞发育潜能的影响[J]. 畜牧兽医学报, 2015, 46(4): 583-591.

GAO Ya-ke，LU Feng-hua，WU Zhu-lian，MA Fan，LIU Xiao-hua，DU Shan-shan，SHI De-shun. Effects of Mitochondrial Transplant on the Developmental Potential of Buffalo Oocytes[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(4): 583-591.

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参考文献

［1］VAN BLERKOM J，DAVIS P，ALEXANDER S.Differential mitochondrial distribution in human pronuclear embryos leads to disproportionate inheritance between blastomeres：relationship to microtubular organization，ATP content and competence［J］.Hum Reprod，2000，15(12)：2621-2633.
［2］VAN BLERKOM J，SINCLAIR J，DAVIS P，et al..Mitochondrial transfer between oocytes：potential applications of mitochondrial donation and the issue of heteroplasmy［J］.Hum Reprod，1998，13(10)：2857-2868.
［3］BRENNER C A，WOLNY Y M，BARRITT J A，et al.Mitochondrial DNA deletion in human oocytes and embryos［J］.Mol Hum Reprod，1998，4(9)：887-892.
［4］ZENG C T，HSIEH R，CHANG S，et al.Pregnancy derived from mitochondria transfer (MIT) into oocyte from patient’s own cumulus granulosa cells (cGCs)［J］.Fertil Steril，2001，76(3):180.
［5］孔令红，刘忠，李红，等.首例经自体颗粒细胞线粒体移植的双胞胎儿出生［J］.第一军医大学学报，2003，(09)：990-991.
KONG L H，LIU Z，LI H，et al.First twins born in Mainland China by autologous granular cell mitochondria transfer［J］.Journal of First Malitary Medical University， 2003，(9)：990-991.(in Chinese)
［6］YI Y C，CHEN M J，HO J Y，et al.Mitochondria transfer can enhance the murine embryo development［J］.J Assist Reprod Genet，2007，24(10)：445-449.
［7］王晓丽，杨娟，陈玉，等.水牛卵丘颗粒细胞凋亡与卵母细胞体外发育潜能的研究［J］.畜牧兽医学报，2009，40(9):1400-1404.
WANG X L，YANG J，CHEN Y，et al.Study on the relationship between cumulus granulose cells apoptosis and oocyte development in buffalo［J］.Acta Veterinaria et Zootechnica Sinica，2009，40(9)：1400-1404.(in Chinese)
［8］华松.体细胞核移植及线粒体命运的研究［D］.杨凌：西北农林科技大学，2007:1400-1404.
HUA S.Studies of somatic cell nuclear transfer and mitochondrial fate［D］.Yangling:Northwest Agriculture and Forestry University，2007:1400-1404.(in Chinese)
［9］高亚可，陆凤花，马帆，等.水牛卵巢颗粒细胞的分离培养及凋亡测定［J］.中国畜牧兽医，2012，39(10)：140-144.
GAO Y K，LU F H，MA F，et al.Isolation，culture and apoptosis determination of buffalo ovarian granulosa cells［J］.China Animal Husbandry & Veterinary Medicine，2012，39(10)：140-144.(in Chinese)
［10］SANTOS T A，SHOURBAGY S E，JOHN J C.Mitochondrial content reflects oocyte variability and fertilization outcome［J］.Fertil Steril，2006，85(3)：584-591.
［11］HANSEN J，CORYDON T J，PALMFELDT J，et al.Decreased expression of the mitochondrial matrix proteases Lon and ClpP in cells from a patient with hereditary spastic paraplegia (SPG13)［J］.Neuroscience，2008，153(2)：474-482.
［12］KRAKAUER D C，MIRA A.Mitochondria and the death of oocytes［J］.Nature，2000，403(6769)：501.
［13］THOUAS G A，TROUNSON A O，WOLVETANG E J，et al.Mitochondrial dysfunction in mouse oocytes results in preimplantation embryo arrest in vitro［J］.Biol Reprod，2004，71(6)：1936-1942.
［14］王晓磊，雷安民，窦忠英.异体颗粒细胞线粒体移植对牛孤雌激活胚胎发育的影响［J］.西北农林科技大学学报，2007，(03)：25-28.
WANG X L，LEI A M，DOU Z Y.Effect of allogranular cells’ mitochondria transfer on bovine parthenogenetic embryos， developmental potencial［J］.Journal of Northwest A&F University，2007，(03)：25-28.(in Chinese)
［15］TAKEDA K，TAKAHASHI S.Dominant distribution of mitochondrial DNA from recipient oocytes in bovine embryos and offspring after nuclear transfer［J］.J Reprod Fertil，2000，116(2):253-259.
［16］HUANG S H，RATLIFF K S，MATOUSCHEK A，et al.Protein unfolding by the mitochondrial membrane potential.［J］.Nat Struct Biol，2002，9(4):301-307.
［17］BLERKOM J V，DAVIS P.High-polarized (ΔΨm HIGH ) mitochondria are spatially polarized in human oocytes and early embryos in stable subplasmalemmal domains：developmental significance and the concept of vanguard mitochondria ［J］.Reprod Biomed Online，2006，13(2):246-254.
［18］BLERKOM J V.Mitochondria as regulatory forces in oocytes，preimplantation embryos and stem cells ［J］. Reprod Biomed Online，2008，16(4):553-569.
［19］YOON S Y，CHA S K，YANG N J，et al.Alteration of high-polarized mitochondria distribution by vitrification of mouse immature oocyte may affect embryonic development［J］.Fertil Steril，2012，98(3):S128-S129.
［20］刘延荷，刘凯，连方，等.卵母细胞线粒体移植对年老小鼠植入前胚胎线粒体膜电位的影响［J］.山东大学学报(医学版)，2009，(01)：6-9.
LIU Y H，LIU K，LIAN F，et al.Effect of mitochondrial transfer on the mitochondrial membrane potential of aging mouse pre-implantation embryos［J］.Journal of Shandong University (Health Sciences)，2009，(01)：6-9.(in Chinese)

线粒体移植对水牛卵母细胞发育潜能的影响

Effects of Mitochondrial Transplant on the Developmental Potential of Buffalo Oocytes

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