早期胚胎体外培养微流控芯片的制备及初步应用

利用微流控芯片模拟输卵管微环境,探讨物理性刺激对早期胚胎发育的影响,为解决早期胚胎体外囊胚发育率较低、胚胎质量差的问题提供帮助。本实验采用一次铸造成型制备微流控芯片培养装置,并应用培养装置对小鼠1-细胞期胚胎进行培养,观察胚胎体外发育率。结果表明,利用微流控芯片培养组与对照组2相比,2-细胞胚胎发育率差异不显著(P0.05),但是,在8-细胞胚胎发育率、桑葚胚发育率和囊胚发育率都差异显著(P0.05)。此外,囊胚内总细胞数显著提高(P0.05)。结论:一次铸造成型微流控芯片制作方法简便易行且培养液不易外漏,这种培养装置能显著提高小鼠早期胚胎体外囊胚发育率和胚胎的质量。     

貉卵母细胞成熟发育的细胞学研究

【目的】探究貉卵母细胞成熟发育特征,为貉卵母细胞的体外成熟培养及优质成熟卵母细胞的挑选提供依据,以提高貉的体外受精效率。【方法】采集1～3岁(性成熟)繁殖期母貉的新鲜卵巢,制备光、电镜样品,观察貉卵母细胞成熟发育过程中细胞器变化;利用X射线微分析技术探索卵母细胞质膜化学元素变化与成熟发育的关系。【结果】根据颗粒细胞层数、卵泡直径、卵母细胞大小以及透明带的变化,将貉卵母细胞成熟发育划分为8个时期。貉卵母细胞在早期发育阶段(第Ⅰ～Ⅳ期)由颗粒细胞包围,逐渐出现透明带、卵泡腔,生发泡内的染色质越来越致密;随着卵母细胞的成熟发育(第Ⅴ～Ⅶ期),微绒毛、皮质颗粒出现;高尔基复合体、线粒体等细胞器数量明显增加,且逐渐迁移至皮质区,同时线粒体、脂滴形态发生变化。卵母细胞成熟时(第Ⅷ期),高尔基复合体及粗面内质网消失,皮质颗粒在质膜下排列;排卵后,核仁发生致密化。随着卵泡腔的增大,卵母细胞质膜表面的钠、氯和硫含量呈下降趋势,第Ⅴ期卵母细胞质膜中钠、氯和硫含量显著高于第Ⅶ期(P<0.05),钙含量显著低于第Ⅵ期和第Ⅶ期卵母细胞(P<0.05);钾含量呈现低-高-低的变化规律,第Ⅵ期卵母细胞质...     

WY14643对小鼠早期胚胎抗氧化损伤的影响

旨在研究WY14643对小鼠早期胚胎发育的影响,探讨WY14643对早期胚胎抗氧化损伤的机理。通过H2O2诱导建立胚胎氧化损伤模型,添加WY14643对胚胎进行体外培养,DCHFDA测定胚胎内部H2O2含量,分别用超氧化物歧化酶(SOD)和微量丙二醛试剂盒测定胚胎不同发育阶段SOD活性和丙二醛MDA含量,并观察后续胚胎发育情况。结果表明:(1)不同浓度WY14643处理2h后,0.1μmol·L-1 WY14643处理组4-细胞率、囊胚率显著高于其他各组(P<0.05);(2)经0.1μmol·L-1 WY14643和H2O2联合处理后,其胚胎发育率显著高于单独H2O2处理组和对照组(P<0.05);(3)DCHFD荧光检测发现,WY14643添加组胚胎内部H2O2水平显著低于其他各组(P<0.05);(4)经SOD、MDA检测,发现添加WY14643胚胎内部各时期SOD水平显著高于其他各组(P<0.05),MDA水平显著低于其他各组(P<0.05)。综上表明,氧化应激对早期胚胎发育是不利的,WY14643通过增加抗氧化酶SOD活性、降低脂质损伤等途径减少胚胎内部ROS,增加抗氧化能力,促进胚胎在体外更好地发育。     

精卵共孵育时间等因素对绵羊卵母细胞体外发育的影响

本试验通过研究精卵作用时间、培养密度、气相环境对卵裂率和囊胚发育率的影响及早期发育快慢与胚胎发育能力的关系,得出:精卵共作用12 h和18 h,不影响囊胚率(P>0.05),但受精18 h的卵裂率明显高于受精12 h的卵裂率(P<0.05);对于四孔板而言,培养密度在50～80枚/孔之间,不影响胚胎后期发育率(P>0.05);早期用5%CO2,空气为平衡气,后期用5%CO2、5%O2,N2为平衡气的气相条件的卵裂率、囊胚率分别极显著高于单独使用这两种气相环境的囊胚率和卵裂率(P<0.01);早期发育较快的胚胎后期发育的能力更强(P<0.01)。     

太湖猪子宫和输卵管冲洗液总蛋白含量的性周期变化

梅山猪(11头),二花脸猪(13头)和梅山×大白杂一代(5头),分成三组,分别测定在性周期不同阶段输卵管和子宫角冲洗液的总蛋白质(OFP和UFP)含量。结果表明:梅山猪二侧OFP平均含量高于二花脸猪和杂种猪,与后者比较差异显著(P<0.05);二侧UFP平均含量,三组比较差异不明显(P>0.05),性周期不同阶段OFP含量,以黄体早期为最高,与黄体后期比较差异显著(P<0.05或0.01);UFP含量亦以黄体早期为最高,与黄体后期比较,二花脸猪差异显著,梅山猪差异不明显。除二花脸猪黄体后期UFP明显低于梅山猪外(P<0.05),OFP和UFP平均含量,三组同期比较差异均不显著。     

猪孤雌激活胚胎的体外培养

试验研究了不同培养体系对猪体外成熟卵母细胞电激活后的体外发育的影响。试验1比较了猪孤雌发育胚胎在NCSU 23、G 2.2添加氨基酸(G 2.2-aa)和G 2.2培养体系中的体外发育效率,结果表明,3个组卵裂率差异不显著(P>0.05),NCSU 23体系的囊胚率显著高于其他2组(14.37±3.77 vs.2.67±2.68和3.13±0.45,P<0.01),但3组间囊胚细胞数差异不显著(21.29±5.27 vs.19.33±2.54和20.27±4.72,P>0.05)。试验2探讨了胚胎培养72h后更换培养液对孤雌发育胚胎的影响,结果显示,培养液更换对卵裂率和囊胚细胞数的影响差异不显著(P>0.05),但更换组囊胚率显著下降(12.50±1.49 vs.5.56±1.89、6.25±1.13、4.64±1.56,P<0.05)。试验3研究了在G 2.2-aa添加胰岛素、亚牛磺酸和半胱氨酸对孤雌激活胚胎发育的影响,结果显示,以上添加剂对卵裂率和囊胚细胞数无显著影响(P>0.05);在G 2.2-aa添加胰岛素、亚牛磺酸和半胱氨酸能明显提高囊胚发育率(6.38±1.00、6.20±2.08、8.73±1.03 vs.2.99±2.21,P<0.05),但以上各组的囊胚率明显低于NCSU 23体系(P<0.05)。结论:NCSU 23是猪孤雌激活发育胚胎较为理想的培养液。     

诱导青年母猪卵泡发育及排卵效果

应用孕马血清促性腺激素(PMSG)制剂对43头青年母猪进行诱导排卵,研究了其卵泡发育及排卵规律,并探讨了诱导处于卵巢机能静止期的母猪发情的最佳途径.结果表明,母猪接受交配时间越长,处理后的排卵效果越好,发情母猪初次交配时间集中在注药后69～126.5 h;卵泡发育总数为(11.90±4.41)个/头,与发情微弱及不发情母猪存在极显著差异(P<0.01),说明卵泡发育总数与母猪发情表现之间存在密切关系;进一步观察青年母猪早期胚胎发育的渐进性,发现进入输卵管内的胚胎处于2～4细胞期,进入子宫时处于4～8细胞期,时间在初配后51～59 h;胚胎全部进入子宫的时间在母猪初配后71～83.5 h,且诱导排卵不影响胚胎的质量.     

交感神经阻断小鼠妊娠早期子宫内肥大细胞的分布

为了研究交感神经对哺乳动物早期胚胎发育的影响机制,通过腹腔注射6-羟多巴胺使交感神经阻断后,观察了小鼠妊娠前期胚胎早期发育和肥大细胞数量和型别的变化。结果显示,交感神经阻断后,不仅对胚胎早期发育有影响,使胚胎着床数降低64.4%,而且影响子宫内肥大细胞的数量及其型别分布。妊娠4d(E4)时肥大细胞数量明显升高(P<0.01),同时肥大细胞分型也有所变化,即黏膜型肥大细胞主要存在于胚胎着床前和着床后,结缔组织型肥大细胞没有明显差异,混合型肥大细胞主要存在于着床期间。这一结果表明,妊娠早期子宫肥大细胞数量与型别的变化可能是交感神经影响早期胚胎发育的途径之一。     

影响黄牛小腔卵泡卵母细胞体外受精及早期胚胎发育的因素

从屠宰场收集黄牛卵巢,取皮质深层卵母细胞进行体外成熟、体外受精和早期胚胎体外培养,分析了影响其效果的因素。结果表明,在成熟培养液中添加FSH(10IU/mL)、HCG(20IU/mL)和17β-E2(1mg/L)对卵母细胞受精后早期胚胎发育能力有极显著促进作用;等量牛卵泡液(BFF)与新生牛血清(NCS)对体外受精胚胎发育效果影响不显著,以15?F为宜;颗粒细胞与输卵管上皮细胞均能显著提高卵母细胞体外成熟受精后早期胚胎的发育率,颗粒细胞 输卵管上皮细胞对克服胚胎阻滞现象效果显著。     

饲粮锌和钙水平对产蛋鸡生产性能、血液生化指标和组织中锌含量的影响

试验在玉米-豆饼型基础饲粮中(含锌29.2ppm)添加不同水平锌和钙研究了饲粮锌和钙水平对蛋用型鸡产蛋期生产性能、血液生化指标和组织中锌含量的影响。结果表明:1.仅喂未加锌的基础饲粮的试验鸡开产晚(P<0.05),产蛋率低(P<0.05)、蛋重小(P<0.05)、产蛋量少(P<0.01)、饲料利用效率差(P<0.01)、破壳率高(P<0.01)。饲粮加锌1000ppm与加锌40ppm对照相比,生产性能无显著差异。饲粮高钙对产蛋鸡生产性能无不良影响。2.血清碱性磷酸酶活性随饲粮加锌水平的提高而显著提高(P<0.01),但不受饲粮钙水平显著影响(P>0.05)。饲粮加锌1000ppm使血清总蛋白含量(P<0.01)、血清白蛋白和球蛋白含量(P<0.05)显著降低。饲粮高钙使血清总蛋白含量(P<0.01)和白蛋白含量(P<0.05)显著降低,而球蛋白含量不受影响(P>0.05)。3.随饲粮加锌水平的提高,肝脏(P<0.01)、胰脏(P<0.01)和胫骨(P<0.01)中锌含量大幅度升高;血液(P<0.01)、肾脏(P<0.01)和肺脏(P<0.05)中锌含量升高幅度较小;而输卵管(P>0.05)、心脏(P>0.05)和肌肉(P>0.05)中锌含量无显著变化。饲粮高钙使肝脏(P<0.05)和肌肉(P<0.01)中锌含量显著降低,对其它器官无显著影响(P>0.05)。     

不同铜源对育成期乌苏里貉生长性能、营养物质消化率及血清生化指标的影响

本试验旨在研究不同铜源对育成期乌苏里貉生长性能、营养物质消化率及血清生化指标的影响。随机选取(60±5)日龄、健康的雄性乌苏里貉60只,随机分为4组,每组15个重复,每个重复1只。在基础饲粮中分别添加硫酸铜(Ⅰ组)、碱式氯化铜(Ⅱ组)、混合铜(Ⅲ组,碱式氯化铜和蛋氨酸铜,添加比例为1∶1,以铜元素计)和蛋氨酸铜(Ⅳ组),添加水平均为40 mg/kg。预试期7 d,正试期55 d。每组选取8只动物进行消化代谢试验。试验结束后,每只动物后肢静脉采血。结果表明:1)Ⅲ组的末重和平均日增重显著高于Ⅰ组(P<0.05),各组间料重比和平均日采食量差异不显著(P>0.05)。2)Ⅱ组的粗脂肪消化率显著高于Ⅰ、Ⅲ、Ⅳ组(P<0.05)。Ⅲ组的铜消化率显著高于Ⅰ、Ⅱ和Ⅲ组(P<0.05),干物质消化率、粗蛋白质消化率各组间差异不显著(P>0.05)。3)粪氮、尿氮含量及氮沉积各组间差异不显著(P>0.05)。Ⅰ、Ⅱ、Ⅳ组的粪铜含量显著高于Ⅲ组(P<0.05)。Ⅳ组尿铜含量显著高于Ⅰ、Ⅱ、Ⅲ组(P<0.05)。Ⅲ组的铜沉积显著高于Ⅰ、Ⅱ、Ⅳ组(P<0.05)。4)Ⅱ组血清中总蛋白含量显著高于Ⅲ、Ⅳ组(P<0.05)。Ⅲ、Ⅳ组血清尿素氮含量显著低于Ⅰ组(P<0.05),其余血清生化指标各组间差异不显著(P>0.05)。在本试验条件下,饲粮中混合添加适宜水平的碱式氯化铜和蛋氨酸铜,可提高育成期乌苏里貉生长性能、营养物质消化率、氮沉积和铜沉积,更有利于其生长发育。     

Preimplantation development of embryos in labrador retrievers

Preimplantation development of canine embryos is not well understood. To understand the timing of preattachment embryogenesis relative to the luteinizing hormone (LH) surge, early embryonic development was examined in Labrador Retrievers after artificial insemination. The embryos migrated from the oviduct to the uterus beginning on day 11 after the LH surge. This transport must be completed within 24 h. By day 13 after the LH surge, all of the embryos had moved and were localized in the uterus. The embryos developed to the morula stage within 11-13 days and to the blastocyst stage within 14 days after the LH surge, respectively. These findings add to the current understanding concerning the physiology of preimplantation development and should help further develop assisted reproductive techniques in canine species, such as cryopreservation and subsequent embryo transfer.     

Effects of purple sweet potato anthocyanins on development and intracellular redox status of bovine preimplantation embryos exposed to heat shock

The development of cleavage stage preimplantation embryos is disrupted by exposure to heat shock, such as high temperatures in the summer season. In this study, we investigated whether addition of anthocyanins, which are strong scavengers of reactive oxygen species (ROS), improves development and intracellular redox status of heat-exposed bovine preimplantation embryos by reduction of heat shock-derived oxidative stress. After in vitro fertilization (IVF), embryos were cultured at 38.5 C through Day 8 (Day 0=day of IVF) with 0, 0.1, 1 and 10 microg/ml anthocyanins (non-heat-shocked group). On Day 2, embryos were cultured at 41.5 C for 6 h with 0, 0.1, 1 and 10 microg/ml anthocyanins followed by culture at 38.5 C until Day 8 (HS group). After exposure to heat shock, the intracellular ROS and glutathione (GSH) contents of individual embryos were measured in the non-heat-shocked and HS groups using fluorescent probes. On Day 8, the blastocysts formation rates of the embryos and total cell numbers of blastocysts were evaluated. Embryos exposed to heat shock without anthocyanins showed a significant decrease in blastocyst formation rate and GSH content (P<0.05) and an increase in intracellular ROS (P<0.05) compared with non-heat-shocked embryos. In contrast, addition of 0.1 microg/ml anthocyanins significantly (P<0.05) improved the blastocyst formation rate of the heat-shocked embryos. Addition of any dose of anthocyanins produced a significant decrease in the ROS levels (P<0.05) and tended to increase the GSH levels under heat-shock conditions. However, addition of higher concentrations (1 and 10 microg/ml) of anthocyanins to the culture media under heat shock did not improve the development of embryos. These results indicate that anthocyanins maintain the intracellular redox balance of heat-shocked bovine embryos by reducing intracellular oxidative stress and increasing the GSH levels. Thus, alterations of the redox state using natural antioxidative polyphenols is a useful approach for reducing heat shock-derived oxidative stress.     

The use of embryonic stem cell derived bioactive material as a new protein supplement for the in vitro culture of bovine embryos

Embryonic stem (ES) cells are expanded versions of the inner cell mass cells that compose the early mammalian blastocyst. Components derived from ES cells may contain various bioactive materials (BM) helpful for early preimplantation embryo growth. In this study, we examined the effect of human ES cell derived BM (hES-BM) on in vitro culture of bovine embryos. When bovine parthenogenetic day 2 embryos were cultured in 10% hES-BM, a significantly higher embryo development rate (44.3%) and increased cell numbers were observed relative to control medium containing 3 mg/ml BSA (19.5%; P<0.01). Among the various concentrations (5, 10 and 15%) and days of treatment (2 or 4 days) tested, 10% hES-BM treatment for 4 days provided the best culture environment to support the growth of bovine embryos in vitro (P<0.05). Little difference was observed between 10% hES-BM and 10% FBS treatment in the examined parthenogenetic or in vitro fertilized embryos, although the hES-BM group developed at a slightly better rate. However, the ICM cell numbers were significantly higher in the hES-BM group in irrespective of embryo origin (P<0.05). In addition, the relative levels of pluripotency (Oct4, × 1.8 fold; Nanog. × 3.3 fold), embryogenesis (Stat3, × 2.8 fold; FGF4, × 18.8 fold; E-cad, × 2.0 fold) and growth (Glut5, × 2.6 fold) genes were significantly higher in the 10% hES-BM group than in the 10% FBS group (P<0.05), while the levels of other genes (Bax, Bcl2, MnSOD and Connexin43) were not different. This is the first report examining the positive effects of hES-BM on bovine embryo development in vitro. Based on our results, we conclude that hES-BM can be used as a new protein supplement for bovine preimplantation embryo development.     

Concentrations of energy substrates in oviduct fluid in unilaterally ovariectomised pigs

Nichol et al (1992, Journal of Reproduction and Fertility, 96, 699–707) identified a pre- to post-ovulatory decrease (approx 1mM) in the amount of glucose in pig oviduct fluid. The present studies investigated whether the decrease was due to metabolism by embroyos and/or oviduct tissues, and also whether there was a local influence of the ovary on the oviduct fluid content of energy substrates. Unilaterally ovariectomised pigs were used, in which, through compensation, oviducts that contained twice the normal number of embryos could be compared with oviducts which contained no embryos. Following unilateral ovariectomy and after two oestrous cycles of normal duration, surgery was performed 88 hours after the beginning of standing heat to obtain oviduct fluid samples, just before embryonic entry into the uterus. Luminal fluid samples from the ampulla and ampullary-isthmic junction from oviducts with and without an adjacent ovary were assayed for glucose, pyruvate and lactate concentrations. No significant differences were found between the glucose, pyruvate and lactate concentrations in fluids from the ampulla or ampullaryisthmic junction from oviducts containing embryos compared with absence of embryos (P> 0·05). Therefore, the post-ovulatory decrease was not due to the presence of embryos or to a local effect of the ipsilateral ovary. Consequently, pig oviduct fluid concentrations of glucose, lactate and pyruvate are seemingly regulated by systemic mechanisms.     

Effects of Exogenous ACTH during Oestrus on Early Embryo Development and Oviductal Transport in the Sow

This study was conducted to assess the effects of ACTH injections on the early development of embryos and their transportation to the uterus. Fifteen sows were monitored for ovulation using transrectal ultrasonography during the first two oestrous periods after weaning. The sows were randomly divided into a control group (C group, n = 8) and an ACTH-treated group (ACTH group, n = 7), and were all surgically fitted with intra-jugular catheters. From the onset of the second standing oestrus after weaning, the sows were injected (NaCl/synthetic ACTH) every 4 h. Blood samples were collected immediately before and 45 min after each injection. All sows were inseminated once 10-33 h before ovulation in their second oestrus after weaning. At 48 (n = 4) or 60 (n = 11) h after ovulation during their second oestrus, the sows were killed and the embryos retrieved from the oviduct and uterus. The embryos were counted and compared with the number of corpora lutea, cleavage rate was noted and, finally, the embryos were prepared for confocal laser scanning microscopy and transmission electron microscopy. There was no difference between the groups regarding cleavage rate, the cytoskeleton, or the number of active nucleoli. However, the ACTH group had significantly (p < 0.05) fewer ova/embryos retrieved (51%) than the C group (81%), and there was a tendency towards faster transportation to the uterus in the ACTH group, possibly because of high progesterone concentrations during treatment. To conclude, administration of ACTH every 4 h from onset of oestrus to 48 h caused significant loss of oocytes or embryos, and possibly faster transportation through the oviduct.     

青年母猪不同生殖器官神经营养因子-4/5的表达研究

本研究旨在探讨神经营养因子-4/5(neuroteophin-4/5, NT-4/5)在青年母猪卵泡期与黄体期卵巢、输卵管及子宫3种生殖器官中表达情况。采用实时荧光定量RT-PCR方法与Western blotting技术对青年母猪卵泡期与黄体期卵巢、输卵管及子宫中的NT-4/5 mRNA与蛋白质表达进行检测。结果表明,NT-4/5 mRNA与蛋白质在卵泡期与黄体期卵巢、输卵管及子宫中均有表达,且卵泡期卵巢、子宫NT-4/5 mRNA表达水平均显著高于黄体期(P＜0.05),而卵泡期输卵管NT-4/5 mRNA表达水平显著低于黄体期(P＜0.05)。试验结果为进一步研究NT-4/5参与青年母猪生殖道生理功能的调节奠定基础。     

玉米赤霉烯酮对青年母猪子宫发育、生长激素分泌及其受体分布与表达的影响

本试验旨在研究玉米赤霉烯酮(ZEA)对青年母猪子宫发育、生长激素(GH)分泌及其受体(GHR)分布与表达的影响。选择胎次和体重[(23.20±0.68)kg]相近的长×大二元青年母猪48头,将其随机分为4组,每组12个重复,每个重复1头。对照组(CON组)饲喂基础饲粮,试验组(T1、T2、T3组)饲喂在基础饲粮中分别添加200、800、1600μg/kg ZEA的试验饲粮。预试期7 d,正试期40 d。结果表明:1)各组间血清及子宫组织中GH含量均无显著差异(P>0.05)。2)各组间子宫组织中ZEA含量均无显著差异(P>0.05)。3)各组间始重、末重、平均日增重(ADG)、平均日采食量(ADFI)及料重比(F/G)均无显著差异(P>0.05);但与CON组相比,T2组子宫指数显著升高(P<0.05),T3组极显著升高(P<0.01)。4)与CON组相比,试验组子宫肌层和内膜增厚,子宫腺数量增多,毛细血管增多,T2和T3组还出现炎性反应及上皮细胞坏死。5)与CON组相比,T2组GHR和Janus蛋白酪氨酸激酶2(JAK2)mRNA相对表达量及T3组GHR mRNA相对表达量显著上调(P<0.05),信号转导与转录激活因子3(STAT3)mRNA相对表达量各组间无显著差异(P>0.05)。6)ZEA对GHR在青年母猪子宫中的分布与定位无明显影响,但与CON组相比,试验组GHR免疫阳性物质分布增多,免疫阳性强度增大。7)各组间子宫GHR蛋白表达量没有显著差异(P>0.05);但与CON组相比,T2组灰度值有升高的趋势(P=0.09)。综上所述,ZEA对青年母猪的生长性能以及血液和子宫中GH和ZEA的含量均无显著影响,但可通过升高GHR、JAK2和STAT3的mRNA相对表达量、增强GHR免疫阳性及升高蛋白表达量来增强GH的生物效应,使子宫发育异常,导致子宫指数出现极显著升高,子宫肌层和内膜增厚,子宫腺和毛细血管数量增多。     

Study on the Key Techniques of Rapid Propagation in Purebred Wagyu

In order to rapid propagation of purebred Wagyu, the experiment were conducted to study the effects of high intensity superovulation, embryo sex identification and different breeds recipient cattle on calving, gestation period and calf birth weight. Wagyu were superovulated repeatedly for nine times with a 30 days interval. The results indicated that the average number of total embryos collected at the second time was 22.78, which was significantly higher than those of the fifth to ninth times (P< 0.05),and there was no significant difference between the other groups (P> 0.05).There was no significant difference between the number of transferable embryos in the first to third times (P> 0.05). The available embryos number of the second times superovulation were significantly higher than those of the fourth to ninth times (P< 0.05),and were remarkably significantly higher than those of the seventh and ninth times (P< 0.01). There were no observably differences of pregnant rate and calving rate between sex identified embryos, and regular embryos respectively (P> 0.05).The female calf percentage of sexed embryos was 95.56%. The calving rate of the Simmental-catalo receptors were higher than those of Wagyu-catalo and Holstein receptors (P> 0.05). The gestation period of Wagyu-catalo were remarkably significantly greater than those of Simmental-catalo and Holstein receptors (P< 0.01), however, there was no significant difference between Simmental-catalo and Holstein receptors (P> 0.05). The birth weight of female calves calved by Simmental-catalo receptors were higher than those of Wagyu-catalo (P< 0.05), which was extremely significantly higher than those of Holstein receptor (P< 0.01). The male calf birth weight was no significant difference in the three breeds receptors (P> 0.05). There were obvious differences between male and female calves in gestation and birth weight with in the same recipient breed.     

Effect of Psammaplin A on in vitro Development of Bovine Aging Oocytes

To investigate the effect of histone deacetylation inhibitor Psammaplin A (PsA) on the development of bovine aging oocytes in vitro,oocytes were randomly divided into control group,aging group and 50 mmol/L PsA treated aging group (PsA group).Immunofluorescence staining and JC-1 were used to detect the blastocyst rate of bovine oocytes after parthenogenetic activation,the number of cells in blastocysts,apoptosis,reactive oxygen species (ROS),glutathione (GSH) and mitochondrial membrane potential intensity of embryos.The results showed that the blastocyst rate of the aging group was significantly lower than that of PsA and control groups (P<0.05).The blastocyst rate of PsA group was not significantly different from that of control group (P>0.05).The number of cells in the blastocysts of control group and PsA group were significantly higher than that of aging group (P<0.05).The number of cells in the blastocysts of PsA group was not significantly different from that of control group (P>0.05).The apoptosis rate in aging group was significantly higher than that of control and PsA groups (P<0.05),the apoptosis rate of PsA group was significantly higher than that of control group (P<0.05).The GSH level of MⅡ oocytes in aging group was significantly lower than that of control and PsA groups (P<0.05).There was no significant difference in GSH level between control and PsA groups (P>0.05).The ROS level of the embryos in aging group was significantly higher than that of control and PsA groups (P<0.05).The ROS level in PsA group was significantly higher than that of control group (P<0.05).The mitochondrial membrane potential of early embryos of aging group 4-8 cells was significantly lower than that of control and PsA groups (P<0.05).The mitochondrial membrane potential intensity of control group was significantly higher than that of PsA group (P<0.05).In summary,PsA could effectively delay the aging of bovine oocytes and improve the quality of oocytes.