卵母细胞

Techniques in embryo technology (such as in vitro production of embryos and animal cloning) need large quantities of high quality oocytes. But the quality of in vitro matured oocytes from slaughtered animals is generally lower than that of the in vivo matured oocytes. It is usually thought that the reason for this poor quality in in vitro matured oocytes is the lack of capacitation during the dominancy of follicular development in vivo.

目前胚胎工程技术研究和开发(如体外生产胚胎和体细胞克隆等)需要大量高质量的成熟卵母细胞，但利用屠宰动物卵巢卵母细胞经过体外成熟培养而获取的卵母细胞质量还远不如体内成熟卵母细胞，其原因一般认为是由于缺乏体内主卵泡阶段的获能作用。

The results showed that mercury chloride (0.5 and 1.5mg/kg BW) had obviously toxicity to the liver and the kidney of mouse, and the dose of 1.5mg/kg BW could damage the ovary function and reduce the number of superovulation oocytes.The results also showed that mercury chloride could inhibit the extruding of the first polar body and affect the quality and the viability of mouse oocyte and reduce the rate of IVF,but little impact on germinal vesicle breakdown of mouse oocyte in vivo.

结果表明，0.5和1.5mg/kg BW汞对小鼠的肝脏、肾脏有明显的毒性作用，1.5mg/kg BW组还对卵巢有明显的毒性作用，并能显著降低超排卵的卵母细胞数；汞对体内卵母细胞生发泡破裂没有影响，但可以抑制卵母细胞第一极体的释放，影响卵母细胞的存活率并降低体外受精率。

The porine is the polyembryony animal, many obtains the ovocyte from the porine ovary although but the maturity quite to be low; Because the ROSI technology is does not have to grow the mature sole round sperm cell to pour into directly completely in the ovicell nature, jumped over the spermatozoon in to pass through the physiology and the biochemistry, if the ovicell nature mature or the activation degree were insufficient, added the round immature sperm cell maturity quite inferior reason, very possibly Causes the ROSI micro fertilization defeat.

猪是多胎动物，从猪卵巢上获得的卵母细胞数量虽然较多，但成熟度比较低；由于ROSI技术是将没有完全发育成熟的单一圆形精细胞直接注入卵胞质内，跳越了精子在穿过透明带和卵质膜等过程中所发生的生理和生化反应；如果卵胞质成熟或活化程度不够，加之圆形未成熟精细胞成熟度比较差等原因，很可能造成ROSI受精的卵母细胞内部激活因子蓄积减少和生成不足，引起精核解聚困难、精圆核不能形成、第二极体不能排出、卵母细胞孤雌发育率增加等，使ROSI显微受精失败。

The new GV preparation method we developed allowed the best visualization of chromatin distribution after Hoechst or CMA3 staining. Our results showed that GV chromatin configurations are different among different species, and thus, while chromatin condensed into a ring around the nucleolus in porcine and bovine oocytes, no such a ring was observed in caprine oocytes; Porcine oocytes were synchronized at GV1, goat oocytes at GV3n and bovine oocytes at GVf configurations before GVBD occurred, indicating that these configurations are progressive towards final maturation rather than atresia; Changes of chromatin configurations were associated with gene activity of transcription; GV chromatin configurations were affected by the environmental changes such as serum starvation and elevated temperature.

应用该技术对猪、山羊和牛不同大小、不同健康程度卵泡及体内和体外成熟过程中卵母细胞染色质构型的变化发现：1）不同动物卵母细胞染色质构型不同；猪和牛卵母细胞染色质围绕核仁凝集形成环，但山羊卵则没有此环。2）卵母细胞在GVBD之前都同步在某一染色质构型；猪同步在GV1，山羊GV3n，牛GVf，说明这些染色质构型代表着进行性变化。3）染色质构型的变化反映基因转录活动的变化。4）染色质构型受环境变化的影响。

Hormone level in maturation medium affected cumulus expansion of COC,but cumulusexpansion became less dependent on hormone level with time of culture.Porcine immaturedoocytes,granulosa cells from small antral follicles and cumulus cells of immatured oocytessecreted cumulus-expansion enabling factors.Granulosa cells from 3-6mm follicles did notproduce CEEF,but there was some CEEF in the follicular fluid of this sized follicles.Sun xingshen, Animal Histology and Embryology

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。

The results showed that : 1 During nonbreeding season, the giant panda′s ovary was still rich with large and small follicular oocytes, and the large follicular oocytes could be matured in vitro; 2 The mean diameter of the oocytes was (141±6.7)μm and lots of cortical granules contained in the cytoplasm of oo...

结果表明∶（1）在非繁殖季节，大熊猫卵巢中小腔及大腔卵泡卵母细胞仍较丰富，其大腔卵泡卵母细胞可在体外培养成熟；（2）大熊猫的卵母细胞平均直径为（141±6.7）μm ，其胞质富含卵黄颗粒；（3）自配 PandaI I号获能液（含丙酮酸钠、肾上腺素等）可使大熊猫精子有效获能；（4）大熊猫卵母细胞体外受精后，经培养可观察到第2 极体，有的受精卵开始发生卵裂。

The results showed that the rates of mature oocyte were 75.2％, 73.1％, 69.8％, 63.5％and oocyte at telophase of MI were 16.3％, 15.9％, 16.9％, 27.0％, respectively. The rates of maturation of oocytes cultured in TCM199 with serum, EGF and TGFαwere significantly higher than that of with BSA team (P＜0.05) and the number of oocyte stayed at telophase of MI in TCM199 with serum, EGF and TGFαteam were significantly less than that with BSA team(P＜0.05); Significant higher rates (66.6％, 66.6％, 73.6％) of normalα-tubulin distribution in oocytes cultured in TCM199+ serum, EGF and TGFαwere compared to that of TCM199+BSA(43.3％)(P＜0.05). The rates of oocyte with cortical granules in cortex were 58.8％, 33.9％, 54.7％and 47.9％respectively, there was significant difference between oocytes cultured with serum, EGF and BSA(P＜0.05). In conclusion, TGFαand EGF can promote the oocyte nuclear transition from telophaseⅠto metaphase of meiosisⅡ, and improved the expression and distribution ofα-tubulin during the ovine oocytes maturation; EGF and serum could promote oocyte cytoplasm maturation. The results suggested that EGF and TGFαmight substitute some substance in serum to improve the quality of oocyte nucleus maturation in vitro, but EGF might be more functional than TGFαto promote the maturation of ovine oocyte ooplasm.

三、EGF和TGFα对卵母细胞成熟的影响未成熟卵母细胞分别在TCM199基础培养液+血清、TCM199基础培养液+BSA+EGF、TCM199基础培养液+BSA+TGFα和TCM199基础培养液+BSA四组成熟培养系统中成熟培养，22小时后上述各组卵母细胞的核成熟率分别为75.2％、73.1％、69.8％、63.5％，处于第一次减数分裂末期的比率分别为16.3％、15.9％、16.9％、27.0％，在添加血清、EGF、TGFα各组中核成熟率显著高于其它各组(P＜0.05)，处于末期的比例明显低于其它各组(P＜0.05)：添加血清、EGF、TGFα各组在a-Tubulin蛋白的分布与表达上(66.6％、66.6％、73.6％)明显高于BSA组的正常率43.3％(P＜0.05)；各组CG发生迁移较好的卵母细胞的比率分别为58.8％、33.9％、54.7％、47.9％，血清组和添加EGF组CG迁移至皮质区的比例明显高于仅添加BSA处理组(P＜0.05)。

The results were as follows: nanos 1 mRNA uniformly dispersed throughout the cytoplasm of oocytes at all stages. Strong expression of nanos1 mRNA was observed in oogonia and stageⅠ、Ⅱ oocytes, but the signal became weaker in later stage oocytes.

结果表明：在斑马鱼卵子发生中，nanos1 mRNA均匀分布于卵原细胞和各时期卵母细胞的胞质中；在卵原细胞和Ⅰ、Ⅱ期卵母细胞中，nanos1 mRNA的杂交信号十分强烈，而较晚期卵母细胞中信号明显减弱。

Stage I of both male and female stone flounder appears only once in all its life, and the germ cells are comprised by spermatogonia or oogonia. The gonad from Mar to Aug keeps at stage II. The gonad index of testis at this stage is 0.037%, and the amount of spermatogonia is increased quickly. There is some linear germ plasm in the cytoplasm of spermatogina. In ovary at this stage it is mostly composed by oocyte of phase 2 which the character is the appearance of yolk nucleus, and no zona radiate in membrane. The mean GI of ovary is 1.95%. From Sep to Oct gonad is at stage III which testis is composed by lots of spermatogina and few spermatocytes, and the mean GI of testis at this stage is 0.086%. In ovary the ooctyes at phase 3 are in dominate position, the yolk nucleus disappear. And the GI of this stage is 3.35%. Both testis and ovary are at stage IV in Nov. hi testis the germ cells are in spermiogenesis, and the mean GI is 0.93%. hi ovary the oocytes are mostly at phase 4, which are filled in the cytoplasm with vitellin granule, and the zona radiate in membrane begins to formation. Nucleus moves to one side of the oocyte gradually. The mean GI of ovary at this stage is 9.37%.

在每年的3月-8月期间性腺处于Ⅱ期，此期精巢中精原细胞明显增多，胞质局部可见有线状的生殖质存在，平均成熟系数为0.037％；卵巢中以2时相卵母细胞为主，可见细胞质中出现强嗜碱性的卵黄核，细胞外由一层滤泡细胞包围，但尚无放射带，平均成熟系数为1.95％。9月-10月期间性腺处于Ⅲ期，此期精巢中仍有大量精原细胞，同时可见部分精母细胞，平均成熟系数为0.086％；卵巢中以3时石鲜孟加限加$玩印面n洲匆s性腺发生、分化及发育的周年变化相卵母细胞为主，细胞质中的卵黄核己消失，平均成熟系数为3.35%。11月性腺处于IV期，此期精巢内精细胞正处于不同的形成过程中，平均成熟系数为0.93%；卵巢中以4时相卵母细胞为主，胞质中充满染成桔红色的卵黄颗粒。

The results showed that after 2 h of culture in MEM the chromosomes of oocyte were seperated to form telophase I while a small spindle was observed around chromosomes of primary spermatocyte. However, two clear spindles were observed in the oocytes cultured in CB containing MEM. After further culture, the chromosomes of both primary spermatocyte and oocyte intermingled and formed one large spindle.

在无CB的培养液中培养的卵母细胞培养2小时后，卵母细胞已经进入第一次减数分裂的后期，染色体开始被拉向两极，而精母细胞的MI纺锤体才刚刚形成，虽然继续培养两者染色体可以合二为一并形成一个纺锤体，但是有些染色体发生滞后；当卵母细胞在含有CB的培养液中培养2小时后，在卵母细胞内形成两个相似大小的MI纺锤体，进一步培养形成一个大的纺锤体，染色体正常。

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Yang yinshu、Wang xiangsheng、Li decang,The first discovery of haemaphysalis conicinna.

1〕 杨银书，王祥生，李德昌。安徽省首次发现嗜群血蜱。