Evaluation of freeze-dried bull spermatozoa by intra-or inter-species micro insemination

通过种内或种间显微授精评价冻干公牛精子

• 批准号: 20580305
• 负责人: HOCHI Shinichi
• 金额: $ 2.91万
• 依托单位: Shinshu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2008
• 资助国家: 日本
• 起止时间: 2008 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20580305/
• 关键词: ウシ精子; 顕微授精; 凍結乾燥; カルシウムオシレーション; DNA脱メチル化; ROCK阻害剤; ガラス化保存; 微小管形成中心; 精子中心体; コメットアッセイ; 胚盤胞; 共焦点レーザー顕微鏡; 超急速ガラス化保存; 卵子活性化

The purpose of this study was to investigate whether freeze-drying process of bull spermatozoa disturbed early post-fertilization events essential to gain the developmental competence of intracytoplasmically sperm-injected(ICSI) oocytes. In interspecies assay using mouse ovulated oocytes, the ability of freeze-dried(FD) bull spermatozoa to induce normal intracellular calcium oscillation pattern was comparable despite of temperatures stored for 1 year. DNA fragmentation in FD spermatozoa stored at+ 4℃was not detected by alkaline comet assay. With intraspecies(bovine-bovine) ICSI, the FD process was found to be innocent in performance of epigenetic remodeling(active DNA demethylation) and function of microtubule-organizing center(sperm aster formation and network assembly) in the pronuclear-stage zygotes. Factors responsible for low blastocyst yield from the FD-ICSI bovine oocytes remain to be determined.

本研究的目的是探讨公牛精子的冷冻干燥过程是否干扰了受精后的早期事件，这是获得卵母细胞胞浆内单精子注射(ICSI)卵母细胞发育能力所必需的。在小鼠排出卵母细胞的种间试验中，冷冻干燥(FD)公牛精子诱导正常的细胞内钙振荡模式的能力是相当的，尽管温度为1年。碱性彗星试验未检测到+4℃保存的FD精子的DNA片段化。对于种内(牛-牛)ICSI，FD过程在原核期受精卵中的表观遗传重塑(活跃的DNA去甲基化)和微管组织中心的功能(精浆形成和网络组装)中被发现是无害的。导致FD-ICSI牛卵母细胞囊胚率低的因素仍有待确定。

项目成果

Stimulatory effect of Rho-associated coiled-coil kinase(ROCK)inhibitor on revivabilitv of in vitro-produced bovine blastocysts after vitrification

Rho相关卷曲螺旋激酶(ROCK)抑制剂对体外牛囊胚玻璃化冷冻后复苏的促进作用

• 发表时间: 2010
• 期刊: Theriogenology
• 影响因子: 2.8
• 作者: S.Hochi;H.Abdalla;H;Hara;M。Shimoda;H.Morita;M.Kuwayama;M.Hirabayashi.;Shinichi Hochi
• 通讯作者: Shinichi Hochi

Vitrification of ICSI- and IVF-derived bovine blastocysts by minimum volume cooling procedure: effect of developmental stage and age

• DOI: 10.1016/j.theriogenology.2010.04.033
• 发表时间: 2010-10-01
• 期刊: THERIOGENOLOGY
• 影响因子: 2.8
• 作者: Abdalla, H.;Shimoda, M.;Hochi, S.
• 通讯作者: Hochi, S.

ウシ卵細胞質内精子注入法の効率改善と凍結乾燥精子を用いた胚盤胞作製の試み

提高牛胞浆内单精子注射效率并尝试利用冻干精子产生囊胚

• 发表时间: 2009
• 作者: Hara;K.;下田美怜;原弘真;下田美怜
• 通讯作者: 下田美怜

Demethylation dynamics of paternal genomes in pronuclear bovine zygotes produced by IVF and ICSI with non-dried or freeze-dried spermatozoa

使用非干燥或冻干精子通过 IVF 和 ICSI 产生的原核牛受精卵中父本基因组的去甲基化动态

• 发表时间: 2009
• 作者: Hara;K.;下田美怜;原弘真;下田美怜;保地眞一;Abdalla H;Abdalla H
• 通讯作者: Abdalla H

In Vitro Fertilization/Intracytoplasmic sperm injection in cattle

牛体外受精/胞浆内单精子注射

• 发表时间: 2011
• 作者: S. Hochi;H. Abdalla;M. Hirabayashi;Shinichi Hochi
• 通讯作者: Shinichi Hochi