共查询到19条相似文献，搜索用时 218 毫秒
1.
《畜牧兽医学报》2015,(4)
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),P0.01),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),P0.01);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(P0.01),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(P0.01)。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%,P0.05),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(P0.05);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(P0.05)。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。 相似文献
2.
Ghrelin对水牛体外受精和孤雌激活胚胎体外发育的影响 总被引：1，自引：0，他引：1
本研究的目的是探讨Ghrelin对水牛体外受精和孤雌激活胚胎体外发育的影响.体外成熟的水牛卵母细胞经体外受精或离子霉素孤雌激活后.分别在舍0,0.5,5,50和500 μg/L Ghrelin的培养液中进行体外培养,观察各组胚胎的卵裂率和囊胚率.结果显示,在培养液中添加不同浓度的Ghrelin对体外受精和孤雌激活胚胎的卵裂率均无显著影响(P>0.05),但添加500 μg/L的Ghrelin显著提高体外受精胚胎的囊胚发育率(33.5% vs 13.7%,P<0.05),50 μg/L或500 μg/L的Ghrelin均显著提高孤雌激活胚胎的囊胚发育率(32.4%和34.6% vs 14.5%,P<0.05).结果表明,培养液中添加Ghrelin对胚胎的早期卵裂没有影响,但可促进水牛体外受精和孤雌激活胚胎囊胚的形成. 相似文献
3.
《畜牧兽医学报》2020,(4)
旨在探讨初次卵裂时间对猪孤雌胚胎发育潜能及其基因相对表达水平的影响。本试验从健康母猪卵巢上抽取卵母细胞进行体外成熟培养,将猪孤雌激活胚胎分为早期卵裂组(16～22 h)与晚期卵裂组(26～32 h)统计比较卵裂率和囊胚率,并对囊胚的多能性相关基因Oct4、Sox2、Klf4等和凋亡相关基因Bcl-xl、Bax、Caspase-3的相对表达水平进行分析检测。结果表明,猪孤雌激活胚胎在16～22 h发生卵裂的为50%～60%,而26 h之后发生卵裂的不到20%,在18 h前完成第一次卵裂的胚胎囊胚发育率为79%,42 h后发生初次卵裂的胚胎无法发育至囊胚期。猪孤雌激活胚胎早期卵裂组的囊胚发育率显著高于晚期卵裂组(P0.05)。早期卵裂组囊胚的Oct4、Nanog、Sox2、Klf4基因的表达量显著高于晚期卵裂组(P0.05),Oct4、Sox2、Klf4基因的相对表达量极显著高于晚期卵裂组(P0.01),Bax和Caspase-3基因的相对表达水平极显著低于晚期卵裂组(P0.01),而Bcl-xl作为保护因子其表达量相对于晚期卵裂胚胎(26～32 h)显著上调(P0.05)。结果显示,初次卵裂时间较早的猪孤雌激活胚胎发育潜能显著高于较晚卵裂胚胎,其囊胚多能性相关基因表达上调,凋亡相关基因表达下调,卵裂时间可作为鉴定猪孤雌激活胚胎发育潜力的重要参数。 相似文献
4.
本研究旨在探究表儿茶素(epicatechin,EC)对小鼠体外成熟培养卵母细胞线粒体DNA(mtDNA)拷贝数及其随后孤雌激活胚胎发育能力的影响。小鼠卵丘-卵母细胞复合体(COCs)在添加不同浓度EC(0、5、10、15、20μmol/L)的成熟液中体外成熟培养16h后,采用实时荧光定量PCR的方法检测卵母细胞mtDNA拷贝数;同时,通过对卵母细胞进行孤雌激活处理,探讨其后续胚胎的体外发育能力。实时荧光定量PCR分析结果显示,添加EC各处理组的卵母细胞mtDNA拷贝数均有所增加,其中,10、15μmol/L组的mtDNA拷贝数均显著高于0μmol/L对照组(P0.05);但10μmol/L组mtDNA拷贝数更接近自然排卵周期合子的mtDNA含量(P0.05)。体外培养观察结果发现,成熟液中添加10μmol/L EC能提高卵母细胞第一极体排出率,与对照组相比差异不显著(P0.05),但能显著提高卵母细胞孤雌激活后胚胎的囊胚发育率(P0.05)。综上表明,小鼠卵母细胞体外成熟液中添加10μmol/L EC可提高卵母细胞的mtDNA拷贝数,有利于促进卵母细胞后续的发育能力。 相似文献
5.
为了研究曲古抑菌素A(TSA)和5-氮杂-2’-脱氧胞苷(5-Aza-CdR)对猪孤雌胚胎发育及胚胎质量的影响,试验采用猪卵母细胞孤雌激活的方法,孤雌激活后在胚胎培养液中分别添加不同浓度TSA和5-Aza-CdR,比较其对猪孤雌胚胎发育的影响。结果表明:40 nmol/L TSA处理24 h能显著提高孤雌胚胎的囊胚率及囊胚细胞个数(P<0.05),卵裂率无明显变化(P>0.05);30 nmol/L5-Aza-CdR处理48 h能显著提高孤雌胚胎的囊胚率(P<0.05),卵裂率与囊胚细胞个数均无明显变化(P>0.05)。说明在一定浓度条件下,TSA和5-Aza-CdR对猪孤雌胚胎发育的囊胚率有显著促进作用,5-Aza-CdR处理对卵裂率、囊胚细胞个数的影响不大,但TSA处理可以明显提高囊胚细胞个数,从而提高胚胎质量。 相似文献
6.
《中国畜牧兽医》2020,(2)
为探讨细胞松弛素B(cytochalasin B,CB)对猪孤雌胚胎和克隆胚胎发育能力的影响,本研究通过在猪体外胚胎培养基中添加不同浓度CB以及不同孵育时间的处理,筛选出CB对猪早期胚胎发育的最适浓度和最佳孵育时间,同时通过Hoechst33342染色检测猪体外囊胚孵化期的细胞数差异,进一步研究CB对孤雌胚胎和克隆胚胎发育的影响。结果显示,培养基中添加CB浓度为7.5μg/mL时孤雌胚胎和克隆胚胎的卵裂率分别为85.00%和90.23%,囊胚率为35.68%和42.58%,均显著高于其他各组(P0.05);采用7.5μg/mL CB处理电激活后的孤雌胚胎和克隆胚胎,孤雌胚胎孵育4 h组的卵裂率(83.80%)和囊胚率最高(35.39%),与其他各组差异显著(P0.05),而克隆胚胎孵育6 h组的卵裂率(83.98%)和囊胚率最高(55.62%),与其他各组差异显著(P0.05)。此外,Hoechst33342染色结果显示,未添加CB处理的孤雌胚胎在囊胚孵化期的细胞平均数为28个,CB处理组的孤雌胚胎和克隆胚胎细胞平均数分别为36和52个,处理组和未处理组细胞数差异显著(P0.05)。结果表明,猪体外孤雌胚胎用7.5μg/mL CB处理4 h可获得较高的卵裂率和囊胚率;体外克隆胚胎用7.5μg/mL CB处理6 h卵裂率及囊胚率最高,且囊胚期内细胞团细胞总数最多。CB处理有利于体外胚胎早期发育,提高克隆胚胎移植受孕率。 相似文献
7.
旨在探讨初次卵裂时间对猪孤雌胚胎发育潜能及其基因相对表达水平的影响。本试验从健康母猪卵巢上抽取卵母细胞进行体外成熟培养，将猪孤雌激活胚胎分为早期卵裂组（16~22 h）与晚期卵裂组（26~32 h）统计比较卵裂率和囊胚率，并对囊胚的多能性相关基因Oct4、Sox2、Klf4等和凋亡相关基因Bcl-xl、Bax、Caspase-3的相对表达水平进行分析检测。结果表明，猪孤雌激活胚胎在16~22 h发生卵裂的为50%~60%，而26 h之后发生卵裂的不到20%，在18 h前完成第一次卵裂的胚胎囊胚发育率为79%，42 h后发生初次卵裂的胚胎无法发育至囊胚期。猪孤雌激活胚胎早期卵裂组的囊胚发育率显著高于晚期卵裂组（P<0.05）。早期卵裂组囊胚的Oct4、Nanog、Sox2、Klf4基因的表达量显著高于晚期卵裂组（P<0.05），Oct4、Sox2、Klf4基因的相对表达量极显著高于晚期卵裂组（P<0.01），Bax和Caspase-3基因的相对表达水平极显著低于晚期卵裂组（P<0.01），而Bcl-xl作为保护因子其表达量相对于晚期卵裂胚胎（26~32 h）显著上调（P<0.05）。结果显示，初次卵裂时间较早的猪孤雌激活胚胎发育潜能显著高于较晚卵裂胚胎，其囊胚多能性相关基因表达上调，凋亡相关基因表达下调，卵裂时间可作为鉴定猪孤雌激活胚胎发育潜力的重要参数。 相似文献
8.
为探讨细胞松弛素B（cytochalasin B,CB）对猪孤雌胚胎和克隆胚胎发育能力的影响,本研究通过在猪体外胚胎培养基中添加不同浓度CB以及不同孵育时间的处理,筛选出CB对猪早期胚胎发育的最适浓度和最佳孵育时间,同时通过Hoechst33342染色检测猪体外囊胚孵化期的细胞数差异,进一步研究CB对孤雌胚胎和克隆胚胎发育的影响。结果显示,培养基中添加CB浓度为7.5 μg/mL时孤雌胚胎和克隆胚胎的卵裂率分别为85.00%和90.23%,囊胚率为35.68%和42.58%,均显著高于其他各组（P < 0.05）;采用7.5 μg/mL CB处理电激活后的孤雌胚胎和克隆胚胎,孤雌胚胎孵育4 h组的卵裂率（83.80%）和囊胚率最高（35.39%）,与其他各组差异显著（P < 0.05）,而克隆胚胎孵育6 h组的卵裂率（83.98%）和囊胚率最高（55.62%）,与其他各组差异显著（P < 0.05）。此外,Hoechst33342染色结果显示,未添加CB处理的孤雌胚胎在囊胚孵化期的细胞平均数为28个,CB处理组的孤雌胚胎和克隆胚胎细胞平均数分别为36和52个,处理组和未处理组细胞数差异显著（P < 0.05）。结果表明,猪体外孤雌胚胎用7.5 μg/mL CB 处理4 h可获得较高的卵裂率和囊胚率;体外克隆胚胎用7.5 μg/mL CB 处理6 h卵裂率及囊胚率最高,且囊胚期内细胞团细胞总数最多。CB处理有利于体外胚胎早期发育,提高克隆胚胎移植受孕率。 相似文献
9.
《动物医学进展》2021,42(8)
为探索哺乳动物孤雌激活胚胎早期发育过程中胞吞作用相关蛋白的动态表达,通过体外生产小鼠孤雌激活胚胎,采用实时荧光定量PCR(RT-qPCR)和蛋白质免疫印迹(Western blot)技术检测不同发育时期小鼠孤雌激活胚胎胞吞作用相关蛋白Cav1、Cav2基因和蛋白相对表达水平,并采用免疫荧光技术进行不同发育时期小鼠孤雌激活胚胎中Cav1、Cav2表达定位分析。结果表明,在小鼠孤雌激活胚胎不同时期均可检测到Cav1、Cav2基因及其蛋白的表达,其中囊胚和桑椹胚中该2个基因的表达水平最高,4-8细胞时期次之,2细胞最低;各发育时期孤雌激活胚胎中Cav1、Cav2蛋白主要定位在胚胎细胞胞质内;囊胚中内细胞团(ICM)细胞中Cav1的荧光强度高于滋养层细胞。说明胞吞作用相关蛋白Cav1、Cav2可能参与小鼠早期胚胎发育的生理调控,桑椹胚和囊胚时期可能是其发挥生理功能的重要时期,并且在囊胚时期对内细胞团发育的调控作用更为显著。研究结果可为探索胞吞作用参与哺乳动物胚胎早期发育的调控机制提供重要参考。 相似文献
10.
试验旨在摸索猪卵母细胞孤雌激活的电场强度和脉冲时间,并探索渗透压阶段培养法对孤雌胚胎后期发育的影响。猪卵母细胞成熟培养42~44 h后,分别在电场强度2.1、2.3、2.5 kV/mm和脉冲时间30、60、90 μs的9组电激活参数下进行孤雌激活试验;卵母细胞在2.1 kV/mm和30 μs的参数下进行孤雌激活后,分别培养于渗透压为271、280、290、302 mOsm的PZM-3中,48 h后移入渗透压280 mOsm的PZM-3中继续培养96 h;孤雌胚胎于电激活后先在含2 mmol/L 6-DMAP的PZM-3中培养4~6 h,然后移入不含6-DMAP的PZM-3中继续培养。试验结果表明,电场强度和脉冲时间两个参数间无显著的交互作用(P>0.05),脉冲时间相同条件下,卵裂率在不同电场强度条件下均无显著差异(P>0.05),2.1和2.5 kV/mm的电场强度条件下,脉冲时间为30 μs时的卵裂率显著高于60和90 μs(P<0.05),而2.3 kV/mm电场强度下3个脉冲时间试验组的卵裂率无显著差异(P>0.05),各试验组的囊胚率无显著差异(P>0.05);孤雌胚胎在渗透压为290~310 mOsm的PZM-3中培养48 h,卵裂率得到显著提高(P<0.05),渗透压对囊胚率无显著影响(P>0.05);6-DMAP对孤雌胚胎卵裂率无显著影响(P>0.05),但可以显著提高囊胚率(P<0.05)。结果提示,猪卵母细胞孤雌激活需要较高的电场强度(2.1~2.3 kV/mm)而脉冲时间不宜过长(30 SymbolmA@s);48 h的高渗培养和6-DMAP的辅助激活有助于孤雌胚胎的后期发育。 相似文献
11.
Takeda K Srirattana K Matsukawa K Akagi S Kaneda M Tasai M Nirasawa K Pinkert CA Parnpai R Nagai T 《The Journal of reproduction and development》2012,58(3):323-329
Interspecies/intergeneric mitochondrial heteroplasmy can occur in interspecies/intergeneric hybrid embryos or following nuclear transfer. In the present study, intergeneric buffalo (Bubalus bubalis) mitochondria (WB-mt) or interspecies murine (Mus spretus) mitochondria (M-mt) were injected into bovine (Bos taurus) oocytes, and the subsequent embryonic development was characterized. Fibroblast mitochondria (WB-mt or M-mt) were microinjected into in vitro matured bovine oocytes followed by oocyte activation by a combination of electrical stimulation and 6-dimethylaminopurine treatment. After seven days of culture, embryo development was evaluated. The copy number of specific mtDNA populations (introduced and native mtDNA) from heteroplasmic oocytes was estimated using real-time PCR. The results illustrated that oocytes injected with either WB-mt or M-mt can develop to the blastocyst stage (20.6% and 19.6%). Cleavage division rates and development to the morula stage in oocytes injected with WB-mt were lower (76.2% and 45.9%, respectively) in comparison with uninjected oocytes (89.2% and 59.1%, respectively) (P<0.05). However, no differences were found in comparing M-mt injected oocytes and controls (P>0.05). An increase in bovine mtDNA copy number was observed at the expanded blastocyst stage of injected embryos (P<0.01), while the number of injected mtDNA was stable throughout development. This study demonstrates that interspecies/intergeneric mitochondrial injected bovine oocytes have the ability to develop to the blastocyst stage after parthenogenetic activation and that injected mtDNA was neither selectively destroyed nor enhanced through development. Moreover, injected intergeneric mitochondria had a demonstrated influence on bovine parthenogenetic development and mtDNA replication. 相似文献
12.
以CZB为基础培养液,培养小鼠4、8-细胞胚胎单卵裂球,研究葡萄糖、牛磺酸和猪输卵管上皮细胞共培养在其体外发育中的作用。结果表明:牛磺酸添加与否对4、8-细胞胚胎单卵裂球的囊胚发育率分别为29%、30%和14%、14%,无显著性差异(P>0.05)。添加葡萄糖后,4、8-细胞胚胎单卵裂球的囊胚发育率分别为36%和19%,有显著提高(P<0.05)。含有葡萄糖而牛磺酸的添加与否对4、8-细胞胚胎单卵裂球的囊胚发育率分别为36%、38%和19%、20%,无显著性差异(P>0.05)。各组内4、8-细胞胚胎单卵裂球形成的囊胚细胞数分别为(10.44±1.24～(12.43±1.18)和(7.57±0.97)～(8.48±1.16),均无显著性差异(P>0.05)。4、8-细胞胚胎单卵裂球与猪输卵管上皮细胞共培养,其囊胚发育率分别为47%和26%,囊胚细胞数分别为(17.57±1.13)和(11.43±0.92),均高于单一培养(P<0.05)。 相似文献
13.
Thongphakdee A Kobayashi S Imai K Inaba Y Tasai M Tagami T Nirasawa K Nagai T Saito N Techakumphu M Takeda K 《The Journal of reproduction and development》2008,54(2):142-147
This study was conducted to investigate the developmental capacity of domestic cat-bovine reconstructed embryos via interspecies somatic cell nuclear transfer (iSCNT) and to observe the mitochondrial DNA (mtDNA) content of the iSCNT embryos. The iSCNT embryos were generated using mixed-breed domestic cat fibroblasts as donor cells and enucleated bovine oocytes as the recipient cytoplasm. When the developmental capacities of iSCNT embryos and parthenogenic bovine embryos were compared, there was no difference (P>0.05) in the rates of cleavage and development to the 8-cell stage (86.6 vs. 84.0% and 32.2 vs. 36.2%, respectively). However, in contrast to development of parthenogenic embryos to the morula and blastocyst stages, no iSCNT embryos (0/202) developed beyond the 8-cell stage. For mtDNA analysis, iSCNT embryos at the 1-cell, 2-cell, 4-cell and 8-cell stages were randomly selected. Both cat and bovine mtDNA quantification analysis were performed using quantitative PCR. The levels of both cat and bovine mtDNA in cat-bovine iSCNT embryos varied at each stage of development. The cat mtDNA concentration in the iSCNT embryos was stable from the 1-cell to 8-cell stages. The bovine mtDNA in the iSCNT embryos at the 8-cell stage was significantly lower than that at the 4-cell stage (P<0.05). No difference in the proportions of cat mtDNA in the iSCNT embryos was found in any of the observed developmental stages (1- through 8-cell stages). In conclusion, bovine cytoplasm supports domestic cat nucleus development through the 8-cell stage. The mtDNA genotype of domestic cat-bovine iSCNT embryos illustrates persistence of heteroplasmy, and the reduction in mtDNA content might reflect a developmental block at the 8-cell stage. 相似文献
14.
Ulloa Ulloa CM Yoshizawa M Yamashita A Hama S Mitsui A Hashi C Abe H Hoshi H Fukui E Matsumoto H 《The Journal of reproduction and development》2008,54(6):465-472
The present study was conducted to determine the criteria for selecting good quality embryos on Day-2 post-insemination and at the blastocyst stage. Bovine oocytes were matured, fertilized and cultured in vitro. First, Day-2 embryos were classified based on the number of blastomeres into 2-cell, 3- to 4-cell, 5- to 8-cell and >8-cell stage embryos; chromosome samples were then prepared. In the second experiment, the Day-2 embryos classified according to the number of blastomeres were cultured separately for an additional 6 days (Day 8). The resultant Day-8 blastocysts from each group of Day-2 embryos were classified into the following 3 grades based on morphology and diameter: Grade A, hatched and hatching blastocysts; Grade B, expanded blastocysts; and Grade C, unexpanded blastocysts. Chromosome samples were then prepared. The 5- to 8-cell stage Day-2 embryos had the lowest incidence of chromosomal abnormalities (13.5%, P<0.05) and the highest development rate to blastocysts (59.2%, P<0.05). Furthermore, the blastocysts derived from the 5- to 8-cell stage embryos had the largest mean number of cells (102.8+/-42.4, P<0.05), largest number of metaphases per blastocyst (9.5+/-4.8, P<0.05) and lowest incidence of chromosomal abnormalities (24.6%, P<0.05). The Grade A blastocysts had the largest mean number of cells (136.6+/-33.4, P<0.05), a large number of metaphases per blastocyst (11.9+/-5.5, P<0.05) and a low incidence of severe chromosomal abnormalities (17.3%). The results showed that, at Day 2, the 5- to 8-cell stage embryos were of better quality since they had the lowest incidence of chromosomal abnormalities and the highest blastocyst rate and the resultant blastocysts had the largest number of cells and lowest incidence of chromosomal abnormalities. In particular, selection of Grade A blastocysts can improve the development rate to term. 相似文献
15.
Zhang K Wei HX Zhang YH Wang SH Li Y Dai YP Li N 《The Journal of reproduction and development》2007,53(3):647-653
The objective of the present study was to investigate the effect of addition of ghrelin to in vitro culture medium on preimplantation development of porcine in vitro fertilized and parthenogenetic embryos. In Experiment 1, we sought to compare the in vitro developmental competence of IVF and parthenogenetic embryos. No significant (P<0.05) differences were detected for cleavage rate or blastocyst rate between the in vitro fertilization (IVF)- and parthenogenetic activation-derived embryos. In Experiment 2, parthenogenetic embryos were cultured in Porcine Zygote Medium-3 containing various concentrations of ghrelin. The blastocyst rate was remarkably (P<0.05) increased when 5 ng/ml (PA-5) and 500 ng/ml (PA-500) of ghrelin was added to in vitro culture medium compared with the other groups. Total cell number per blastocyst was slightly promoted in the ghrelin treatment groups compared with the controls. However, the ratio of inner cell mass (ICM) cell number/total cell number was significantly reduced in the PA-50 group compared with the controls (P<0.05). In Experiment 3, we cultured in vitro fertilized embryos in Porcine Zygote Medium-3 supplemented with ghrelin at different dosages. The rate of blastocyst formation was markedly (P<0.05) elevated when 500 ng/ml ghrelin was added to culture medium (IVF-500) compared with the controls. Increased total cell numbers (P<0.05) were observed when in vitro fertilized embryos were cultured in IVF-50 and IVF-500 compared with the controls. However, the ratio of ICM cell number/total cell number was decreased in the ghrelin treatment groups compared with the controls (P<0.05). Taken together, the results suggest that ghrelin can enhance blastocyst formation of porcine in vitro fertilized and parthenogenetic embryos while exerting a negative effect on the structural integrity of the blastocysts. 相似文献
16.
This study was conducted to improve the developmental ability of nuclear transfer (NT) embryos by using blastomeres from in vitro fertilized (IVF) embryos with high quality as donor cells. The IVF embryos selected at the 2-cell stage at 24-h postinsemination (hpi) and again at the ≥8-cell stage at 48 hpi (Selected-IVF-embryos) showed the highest blastocyst formation rate among embryos. When blastomeres from the Selected-IVF-embryos (Selected-NT group) or Nonselected-IVF-embryos (Non-selected-NT group) were used as donor cells for NT, the blastocyst formation rate in the Selected-NT group (25.6%) was significantly higher than that in the Non-selected-NT group (13.5%). When blastomeres from the Selected-IVF-embryos at 108 (contained many cells before cell division) and 126 hpi (contained many cells immediately after cell division) were used as donor cells for NT (108- and 126-NT groups, respectively), the 126-NT group showed a significantly higher blastocyst formation rate (32.1%) than the 108-NT group (16.8%). Embryo transfer of blastocysts in the 126-NT group showed that 11 of 23 recipients became pregnant; nine calves were obtained. For the NT embryos reconstructed using in vivo derived embryos, 9 of 20 recipients became pregnant; seven calves were obtained. These results indicate that the blastocyst formation rate of NT embryos can be improved by using blastomeres from IVF embryos selected at the early developmental stage, especially immediately after cell division, and that the resultant NT embryos have a high developmental ability to progress to term that is comparable to NT embryos reconstructed using in vivo derived embryos. 相似文献
17.
Thanh Quang Dang‐Nguyen Nguyen Viet‐Linh Tamas Somfai Seiki Haraguchi Bui Xuan Nguyen 《Animal Science Journal》2014,85(5):517-523
In the present study, we examined the development to blastocysts of large and small blastomeres from unevenly cleaved 2‐cell embryos (uneven 2‐cell embryos) in pigs. Proportion of blastocysts derived from large blastomeres (52.8 ± 6.4%) was significantly higher (P < 0.05) compared with small ones (32.1 ± 4.6%). However, there were no differences in total cell number, inner cell mass (ICM) cell number and ICM/total cells ratio between them. Of 53 sister blastomere pairs in the same embryos examined there were 12 pairs (22.6%) in which both blastomeres developed to blastocysts, 16 pairs (30.2%) in which only large blastomeres developed to blastocysts, and five pairs (9.4%) in which only small blastomeres developed to blastocysts. Relative total amount of active mitochondria in small blastomeres were lower (P < 0.05) than that of large blastomeres and blastomeres from evenly cleaved 2‐cell embryos. However, there was no difference in relative density of active mitochondria in these three types of blastomeres. In conclusion, blastocysts derived from small and large blastomeres in uneven 2‐cell embryos had comparable quality in terms of cell number, ICM number, ICM/total cell ratio and distribution of active mitochondria. The results suggest that these blastomeres may contribute multiple offspring production in pigs. 相似文献
18.
为研究持续不同时间的冷、热应激对猪孤雌胚胎体外发育的影响,本研究以猪孤雌胚胎为材料,采用免疫荧光染色、实时荧光定量PCR技术检测不同时间的冷（31℃）、热应激（41℃）处理对猪孤雌胚胎发育后囊胚发育率、细胞数、细胞凋亡率、自噬相关基因及细胞凋亡相关基因mRNA转录水平的影响。结果显示,热应激12 h后囊胚发育率显著低于对照组（P<0.05）,冷应激18 h后囊胚发育率显著低于对照组（P<0.05）,而冷应激组囊胚发育率高于热应激组。热应激12 h和冷应激18 h后均导致囊胚内细胞数显著低于对照组（P<0.05）,细胞凋亡率显著高于对照组（P<0.05）,且冷应激组的细胞凋亡率低于热应激组。冷、热应激组自噬相关蛋白LC3的表达均高于对照组;冷、热应激中自噬相关基因Atg6和Atg8的表达均极显著高于对照组（P<0.01）,Lamp2基因的表达均显著高于对照组（P<0.05）,热应激组中的Atg6和Atg8基因的表达高于冷应激组。通过检测细胞凋亡相关基因mRNA的转录水平发现,冷、热应激组中细胞凋亡相关基因Bak、Casp-3、Fas的表达均极显著高于对照组（P<0.01）,Bcl-xl基因的表达均显著低于对照组（P<0.05）。综上,猪孤雌胚胎对冷应激（31℃）的耐受性比对热应激（41℃）强,且热应激可诱导体外培养的猪孤雌胚胎自噬及凋亡相关基因的表达,从而降低孤雌胚胎发育的能力。 相似文献
19.
试验旨在探究高浓度葡萄糖对猪卵母细胞体外成熟及早期胚胎发育能力的影响。取体外分离处于生发泡期的猪卵丘卵母细胞复合体(COCs),分为3个处理组。分别用含葡萄糖浓度为5.6 mmol/L(C组)、10 mmol/L(G-1组)、15 mmol/L(G-2组)的培养液,进行体外成熟(IVM)处理,42 h后观察,并统计卵丘细胞扩散情况和第一极体排出率;对体外成熟42 h后的卵母细胞孤雌激活,统计2-细胞、4-细胞和第7天囊胚发育。结果发现,G-1组和G-2组卵丘细胞扩散度显著低于C组(P<0.05);G-1组和G-2组的MII期卵母细胞死亡率和存活率与C组相比无显著差异(P>0.05),但G-1组极体率显著降低(P<0.05),G-2组极体率极显著低于C组(P<0.01)。孤雌激活后,与C组相比,G-1组和G-2组的2-细胞分裂率显著降低(P<0.05),4-细胞分裂率以及囊胚发育率均极显著降低(P<0.01),但G-1、G-2组囊胚细胞数量与C组相比无显著性差异(P>0.05)。进一步线粒体染色发现,G-1组和G-2组的线粒体与C组相比分布不均。与C组相比,荧光结果显示G-1组和G-2组不仅活性氧(ROS)水平极显著升高(P<0.01),而且G-1组与G-2组谷胱甘肽(GSH)含量显著降低(P<0.05),虽然G-1组丙二醛(MDA)无显著性差异(P>0.05),但G-2组丙二醛(MDA)含量显著升高(P<0.05)。研究结果表明,葡萄糖浓度高会影响猪卵母细胞线粒体分布,氧化应激水平升高,成熟效率降低,损害早期胚胎的发育潜能。 相似文献