卵母细胞
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Japonicus and analyzed the role of animal process of oocyte during ovulation and oocyte maturation. What's more, we studied the role of microtubules and microtubule organizing center upon GV migration. The oocyte of A. japonicus connects to the follicle via animal pole process. When placed in normal sea water, ovulation started spontaneously.
利用DTT诱导刺参卵母细胞成熟,综合运用光镜、电子显微镜及激光共聚焦技术观察卵母细胞成熟过程,分析了卵母细胞表面动物极突起在卵母细胞脱滤泡中的作用,以及卵母细胞动物极突起和微管组织在生发泡移动过程中所起的作用。
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The results showed that 1 there were no significant differences in the rates of cytoplast protrusion and enucleation between oocytes that were incubated in colchicine (0.4 μg/mL) for 0.5 h and oocytes that were incubated in colchicine (0.4 μg/mL) for 1 h, and the rate of cytoplast protusion can be 85.4% while the rate of cytoplast enucleation is 100%. 2 There was no significant difference in oocyte enucleation between oocytes treated with medium containing 0.2 μg/mL colchicine for 0.5 h and oocytes treated with medium containing 0.4 μg/mL colchicine for 0.5 h. 3 A maturation time of 18–23 h did not affect the rates of cytoplast protusion and enucleation by chemically assisted enucleation, whereas the rate of enucleation of oocytes by blind enucleation was found to decrease with a prolonged incubation time. 4 The development rates of reconstructed embryos could not be influenced by these two enucleation methods, increased from oocytes matured for 21–23 h.
结果表明: 1 卵母细胞在0.4 mg/mL的秋水仙素溶液中分别孵育0.5 h和1 h,胞质突起率和去核率没有显著的差异,突起率可高达85.4%,去核率达到100%; 2 0.2 mg/mL或0.4 mg/mL秋水仙素溶液将卵母细胞处理0.5 h,对去核效果没有显著影响; 3 对于体外成熟18~23 h的卵母细胞,随着成熟时间的延长,盲吸法的去核率降低,但没有影响秋水仙素诱导胞质突起的比率和去核率; 4 两种去核方法对重构胚的发育没有产生显著影响,但成熟21~23 h卵母细胞重构胚囊胚的发育率显著高于成熟18~20 h卵母细胞重构胚囊胚的发育率。
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We detected that EGF mRNA was expressed sflungly lii the oocyte, and is also found hi gmnulosa cells, the cell fium smaller foflicular expressed stronger than fium bigger one. In the corpus hemonbaglcwn corpus luteurn, lean type and pseudocorpus-luteum, EGF rnRNA was detected,, no distinct difference can be seen in them. The EGF mRNA expressed strongly in fimbria end, ampulla and isthmus of oviduct, in the big follicular stage, ovulation stage, pregnancy stage and spurius pregnancy stage, we can not see any distinct change in them, but hi the medium follicuar stage,it is weaker.
结果发现:猪卵母细胞中EGF的mRNA强烈表达,且小卵泡卵母细胞→中卵泡卵母细胞→大卵泡卵母细胞中,EGF的mRNA表达量有逐渐减少的趋势;猪卵泡的颗粒细胞中有EGF的mRNA表达,小卵泡颗粒细胞→中卵泡颗粒细胞→大卵泡颗粒细胞中,EGF的mRNA表达也有逐渐减少的趋势;猪卵巢中的红体、黄体、白体和假黄体中都有EGF的mRNA表达,看不出几部分的表达量有明显的强弱变化;猪输卵管伞部、壶腹部和峡部,都有EGF的mRNA表达,在大卵泡期,排卵期,孕期和假孕期都强烈表达,各期间看不出明显的强弱变化,中卵泡期表达较弱;猪子宫中EGF的mRNA在大卵泡期,排卵期,孕期和假孕期都强烈表达,看不出表达量的明显变化,而小卵泡期表达量明显减弱。
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Hormone level in maturation medium affected cumulus expansion of COC, but cumulus expansion became less dependent on hormone level with time of culture. Porcine immatured oocytes, granulosa cells from small antral follicles and cumulus cells of immatured oocytes secreted cumulus-expansion enabling factors. Granulosa cells from 3-6mm follicles did not produce CEEF, but there was some CEEF in the follicular fluid of this sized follicles.
激素水平对卵丘扩展有影响,但随培养时间的延长,对激素的要求有逐步减少的趋势;猪卵丘扩展不依赖于卵母细胞,卵母细胞核成熟也与卵丘扩展无关;GV期卵母细胞、小腔卵泡内颗粒细胞和成熟卵的卵丘细胞都有较高的分泌CEEF能力;3-6mm卵泡的壁颗粒细胞不能分泌CEEF,但3—6mm卵泡的卵泡液中有CEEF。
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In conclusion, these results suggested that (1) the ageing-associated decline in fertility of female KM mouse was due to a decrease in both quantity and quality of oocytes;(2) the ooplasm from young mice did not correct the meiotic errors of aged mice, and the ooplasm from aged mice did not induce abnormal segregation of meiotic chromosome of young mice, indicating that meiotic anomalies found in the oocytes of aged mice might be related to nucleus or chromosomes and relevant factors;(3) a critical nucleocytoplasmic ratio was essential for normal maturation and segregation of meiotic chromosomes of oocytes and development to 2-cell embryos.
本研究结果表明:(1)昆明白小鼠与衰老相关的生育力下降是其卵母细胞数量减少和质量下降的综合结果;(2)年轻小鼠卵母细胞细胞质不能纠正老龄小鼠GV的减数分裂错误,老龄小鼠卵母细胞细胞质也不诱导年轻小鼠的GV发生减数分裂错误,老龄小鼠卵母细胞减数分裂异常很可能与细胞核或染色体及其相关因素有关;(3)关键的核质比对卵母细胞正常的成熟、减数分裂染色体分离及2-细胞期胚的发育是绝对必需的。
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The result of frozen MⅡoocytes using three methods indcated that the self-restraintspear method was better than OPS, and the effection of frozen altogether with self-restraintspear method and centrifugal was the best; the frozen effection was not ideal using fluidspear for 8- cell ovocyte.
用三种冷冻方法(OPS法;自制移液枪头法;移液枪头法+离心法)对MⅡ卵母细胞冷冻的结果表明,自制移液枪头法优于OPS法,其中以移液枪头法+离心组的冷冻效果最好;用移液枪头法对体外发育到8-细胞期卵母细胞的冷冻效果不理想。MⅡ期卵母细胞用OPS法冷冻后卵母细胞IVF和电激活的卵裂率都显著高于移液枪头法(P<0.01),电激活后2-细胞卵裂率比IVF高。
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To pronuclear formation rate after oocyte in vitro maturation 18 hours and parthenogenticactivation, oocytes derived from SM is significantly higher than oocytes from PM,oocytes derived from IM is significantly lower that of oocytes detrived from PM and SM.
对于孤雌刺激反应而言,无论是何种类型的卵母细胞成熟18小时后,自发成熟卵母细胞激活后形成原核的比率高于被动成熟的卵母细胞,诱导成熟的卵母细胞的原核形成率显著低于自发和/或被动成熟的卵母细胞。
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Under the transmission election microscope, we had observed that the state of oocytes arid the cumulus cells around them was tightness, and there were plenty of long and thin microvilli playing the role of connection in the enwrapping. The shape of cumulus cells nucleus was ovate, and the longer and shorter diameters of the cumulus cells were 4.250±0.042μm and 2.750±0.07 μm respectively. The zona pellucida was well proportioned and tightly linking with oocyte. There were great deals of multi-crista mitochondria, lipid droplet and vacuolus. The mitochondria longer and shorter diameters were 0.600±0.106μm and 0.490±0.117μm. Granular nucleoplasma distributed very well in the cell nucleus.
电镜下,卵母细胞外的卵丘细胞紧紧包裹卵母细胞,且与卵母细胞之间有大量细长的微绒毛相联系;卵丘细胞核呈卵圆形,其长径为4250±0.042μm,短径为2.750±0.071μm;透明带均匀,与卵母细胞结合紧密;在皮质区集中分布大量的多峰型线粗体、脂滴和空泡,线拉体长径为0.600±0.106μm,短径为0.490±0.117μm;细胞核中颗粒状的核质分布非常均匀。
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Cell-cycle synchronization between the donor cell and recipient oocyte determines the embryo development in nuclear transfer. In the present study, we microinjected primary spermatocyte into the perivetelline space of oocyte. 37% pairs fused after electric stimuli and the resulting oocytes were culture for 2 h in MEM with or without CB.
在本研究中,我们将小鼠的初级精母细胞显微注入MI卵母细胞的透明带下,经直流电脉冲作用后有37%的初级精母细胞融入卵母细胞,然后将融合的卵母细胞分成两组在MEM和含有CB的MEM培养液中分别培养,2小时后将在CB培养液中培养的卵母细胞转入正常培养液中。
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However, when two maturing oocytes fused and two spindle will form in the big cell, the chromosomes will not intermingle. In the present study, we removed GV from one oocyte and transfer to the perivetelline space of another GV oocyte. After fusion the resulting oocytes which contained two GVs were cultured further in MEM.
在本实验中,我们利用小鼠GV期卵母细胞将一枚卵母细胞的生发泡取出后移至另一未经去核处理的GV期卵母细胞透明带下,经三次电脉冲作用后将融合的含有两个GV的卵母细胞放入MEM成熟培养液中培养,在培养成熟的不同阶段分别收集卵母细胞进行免疫荧光染色观察微管及核的变化。
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