共查询到17条相似文献，搜索用时 359 毫秒
1.
用2种不同的配方分别对猪GV期和MⅡ期卵母细胞进行玻璃化冷冻研究,并用孤雌激活的方法检测冷冻-解冻对卵母细胞再发育能力的影响。结果如下:(1)以PBS和TCM1992种配方为冷冻基础液,分别对猪GV期和MⅡ期卵母细胞进行玻璃化冷冻,解冻后对细胞进行体外成熟观察、形态学观察、染色观察,差异不显著;(2)经冷冻-解冻,猪GV期卵母细胞比MⅡ期卵母细胞成活率高、孤雌激活效果好;(3)经冷冻-解冻,猪卵母细胞再发育能力明显低于新鲜细胞,冷冻过程对卵母细胞的发育能力造成了明显的影响。 相似文献
2.
在不同的外界因素下,使用胞质内体细胞核注射的方法,为进一步提高猪体细胞核移植的效率来寻找新途径。利用屠宰场卵巢采集的卵母细胞经过去核,注核及激活等操作后,研究猪颗粒细胞和胎儿成纤维细胞,血清饥饿处理法及不同的激活时间等因素对猪核移植重构胚体外发育的影响.结果表明,猪颗粒细胞和胎儿成纤维细胞用作核供体与猪卵母细胞构建重构胚,在卵裂率上无显著性差异。作为供核细胞的猪成纤维细胞,经过血清饥饿与不经过血清饥饿培养,核移植重构胚的卵裂率无显著性差异。在进行核注射后,使供核细胞与胞质受体作用适当的时间(1～6h)在进行激活,有利于重构胚的发育。提示颗粒细胞和猪胎儿成纤维细胞用作核供体与猪卵母细胞构建重构胚以及作为供核细胞的猪成纤维细胞,经过血清饥饿与不经过血清饥饿培养处理后,这二者对猪核移植重构胚发育的均无明显影响。但是使供核细胞与胞质受体作经过适当的时间(1～6h)的激活,却有利于重构胚的发育。 相似文献
3.
本试验探讨了不同基础培养液(mTCM-199、NCSU23)和PMSG、HCG浓度、作用时间对猪卵母细胞体外成熟的影响和乙醇及乙醇与各种化学试剂联合处理对猪体外成熟卵母细胞孤雌激活、孤雌生殖胚胎发育的影响,最终找到了最佳的猪卵母细胞体外成熟培养体系,并确定了乙醇孤雌激活的最佳浓度、作用时间和与不同化学试剂联合处理的最佳组合。试验结果得出:最佳的猪卵母细胞培养体系为:NCSU23+20IU/mlPMSG+20IU/mLHCG+10%PFF,激素作用时间为45h,成熟率可达到70.3+3.9%;乙醇的最佳激活浓度为8%,激活率为29.6+2.5%;乙醇激活的最佳处理时间为10min,激活率为32.6+4.1%;乙醇与CB+CHX联合处理的激活率最高,激活率为60.8+3.2%,卵裂率、3～4细胞率、8细胞及8细胞以上率分别为45.1+3.0%、28.6+3.7%、8.7+1.2%;乙醇与CB+6-DMAP联合处理的激活率为51.7+1.9%,卵裂率、3￣4细胞率、8细胞及8细胞以上率分别为41.7+3.6%、30.3+4.3%、9.9+1.8%,虽然激活率、卵裂率低于乙醇与CB+CHX处理组,但3￣4细胞率、8细胞及8细胞以上率均高于乙醇与CB+CHX处理组(30.3+4.3%vs28.6+3.7%,9.9+1.8%vs8.7+1.2%)。因此,乙醇激活的最佳方法为:8%乙醇处理10min,CB+6-DMAP处理7h。 相似文献
4.
本实验主要比较了离子霉素、电脉冲2种方法激活牛、羊体外成熟卵母细胞的效率。2种激活方法中,牛胚胎卵裂率无显著差异(90.61%VS94.40%,P>0.05),而离子霉素激活胚胎的囊胚发育率极显著高于电激活方法(12.3%VS2.4%,P<0.01)。2种方法对羊的胚胎的研究中,羊胚胎卵裂率无显著差异(72.4%VS77.4%,P>0.05)。但是离子霉素激活胚胎的囊胚发育率显著高于电激活方法(3.67%VS10.4%,P<0.05)。本实验中还比较了用化学激活(离子霉素)激活牛体外成熟卵母细胞后,用SoFaa体系培养,换液与不换液对孤雌激活胚胎体外发育的影响。结果表明:在第四天不换液的胚胎卵裂率和囊胚率极显著高于换液的胚胎(11.64%VS3.49%,P<0.01)。 相似文献
5.
6.
7.
介绍了以屠宰场废弃的猪卵巢为材料,通过系统技术途径得到猪早期胚胎的基本方法。新鲜的猪卵巢采集后,25～35℃条件下保存,在3h内进行卵泡卵母细胞与卵丘细胞复合体(COCs)采集与分级,对COCs中卵母细胞体外成熟(IVM)、体外受精(IVF)及其受精卵体外发育(IVD)等关键技术环节进行了系统地探索,重点探讨了不同发育阶段(直径大小)卵泡卵母细胞、不同培养系统与添加成分以及不同培养时间等关键因素对猪卵母细胞IVM的影响,并对其IVF卵裂能力进行了比较与评价。实验结果表明,采用mTCM199作为基础成熟培养系统液,并添加ф5～8mm卵泡液(pFF)和雌二醇(E2),ф2～8mm卵巢卵泡COCs经IVM40h可得到较高的成熟率(66.7%)和IVF卵裂率(29.2%)。本研究初步建立了猪2-4-细胞早期胚胎的体外批量生产技术,可望进一步改进和完善,应用于养猪业科研与生产实践。 相似文献
8.
用海星鞘神经提取液和二硫苏糖醇对刺参卵母细胞进行了体外人工促熟研究。结果显示,二硫苏糖醇对刺参卵母细胞具有明显的促熟作用。刺参卵母细胞包被在滤泡中发育,卵母细胞通过动物极的卵突起连接到单层细胞构成滤泡膜上,充分发育的卵母细胞处于生发泡期,染色质凝集在核仁周围,处于生发泡期的卵母细胞在海水中不会自发成熟。用二硫苏糖醇处理充分发育的卵母细胞20min,卵母细胞的生发泡开始移动,并锚定在卵母细胞的动物极,继而出现生发泡破裂。此后,减数分裂重新启动,第一、二极体分别排出。但海星鞘神经提取液对刺参卵母细胞没有促熟作用。实验表明,二硫苏糖醇浓度在10-5～10-1mol/L范围内,均可促使刺参卵母细胞发生生发泡破裂,10-2mol/L时诱导卵母细胞成熟率可达90%以上。 相似文献
9.
10.
11.
采用光镜和透射电镜技术观察多刺裸腹溞（Moina macrocopa）孤雌溞卵子的发生过程。根据卵母细胞细胞核的形态、卵黄的合成、积累情况及核物质的形态变化等观察结果,将卵子发生分为3个时期：卵原细胞期、卵母细胞期和成熟卵母细胞期。卵原细胞核大,胞质层薄,有少量线粒体分布,核物质浓缩为一团;卵母细胞发育的早期和中期细胞体积增大,细胞质层增厚,线粒体迅速增多,细胞核变圆,核物质分散成多块;后期卵母细胞胞质中开始出现大量卵黄颗粒和少量脂滴,细胞核边缘波浪化,线粒体空泡化,其中卵黄合成初期,核物质重新聚集在细胞核中央,在卵黄合成中期又分散开来;成熟卵母细胞质中充满卵黄颗粒,呈强嗜酸性,细胞核的形态呈不规则状,核物质分散。多刺裸腹溞生殖腺的外部形态随着生殖周期的变化而不同。在孤雌溞的整个生活史中生殖腺内卵子的发育经历了“同步一不同步一局部同步”的发育模式。发现1龄幼溞生殖腺中不存在卵原细胞,成龄溞的生殖腺有多个发育时期的生殖细胞并存。同时在亚显微水平上发现卵黄主要由线粒体演变而来,也通过吞饮作用吸收成熟的卵黄颗粒,内源与外源性共同作用形成卵黄。[中国水产科学,2006,13（4）：547—554] 相似文献
12.
Juliana C. Veit Jaqueline Piccolo Grasiela Facco Aline F. Scherer Bernardo Baldisserotto Gessi Koakoski 《Journal Of Aquatic Food Product Technology》2017,26(3):308-324
Behavioral responses, blood markers, and fillet properties were investigated after silver catfish exposure to different electric field strengths, frequency, and duration of electric current in the preslaughter stunning. All combinations of electric fields and frequencies were able to stun fish. Longer apparent stun was obtained at the intermediate electrical frequencies. High electric field strength detracted the texture of fillets. At least 5 s was required to stun fish, and longer exposure to electrical current did not prolong the apparent stun or damage fillet properties. Results indicate that silver catfish are relatively resistant to electronarcosis. 相似文献
13.
大口鲶(Silurus meridionalis)是一种古老的具有电感受器官的鱼类,其头部分布着大量微壶腹状的电感受器官,可以感受到生存环境中的微弱电信号,并在其摄食、求偶等行为中具有重要作用。本实验采用行为学的方法研究了大口鲶在不同直流类匀强电场下的反应特征。结果表明,在输出电压为0.9V、水中电场强度为(5.0±0.1)mV.cm-1时,大口鲶对铜板电极表现出最大的偏好性,在电场范围停留时间可达100 s以上,说明弱电场对大口鲶具有较强的吸引。研究结果为采用弱电诱集大口鲶提供了科学依据,并为改进传统的大口鲶捕捞作业方式及新渔具渔法的研制开发提供参考。 相似文献
14.
Fertilizing ability of gametes at different post‐activation times and the sperm–oocyte ratio in the artificial reproduction of pikeperch Sander lucioperca 
下载免费PDF全文
下载免费PDF全文 Jiri Kristan Daniel Zarski Miroslav Blecha Tomas Policar Oleksandr Malinovskyi Azin Mohagheghi Samarin Katarzyna Palinska‐Zarska Joanna Nowosad Slawomir Krejszeff Dariusz Kucharczyk 《Aquaculture Research》2018,49(4):1383-1388
The time period during which oocyte and spermatozoa retain their fertilizing ability after contacting with water was evaluated in pikeperch (Sander lucioperca). In addition, success of in vitro fertilization was examined regarding to the sperm‐to‐oocyte ratio (SOR). In the first trial, oocytes were placed in Petri dishes containing 5 ml of the hatchery water, to which freshly collected and pooled sperm were added to each sample at 0, 15, 30, 60, 90, 120, 150 and 180 s post oocyte activation. The oocytes retained their fertility for at least 30 s after contacting with water. The second trial tested the maximum time period during which spermatozoa retained fertilizability after contacting with water. Milt (50 μl) was collected from each male and added to 5 ml of water in Petri dishes. Thereafter, oocytes were added at 0, 5, 15, 30, 60 and 75 s post‐sperm activation. Delays exceeding 10 s affected negatively the fertilization success. The third trial examined the optimum SOR; in which was found that 100 × 103 spermatozoa per oocyte were the minimum ratio to ensure fertilization rates above 70%. Overall, the data clarified some biological interactions of gametes in the artificial propagation of pikeperch. 相似文献
15.
P. Mark Lokman Kataraina A.N. George Graham Young 《Fish physiology and biochemistry》2003,28(1-4):283-285
To investigate the hormonal control of previtellogenic oocyte growth in eel, Anguilla australis, ovarian explants were incubated with or without different doses of steroid (11-ketotestosterone, estradiol-17β) or peptide hormones (ovine growth hormone, human insulin-like growth factor-I, human chorionic gonadotropin, porcine insulin) for 18 days. 11-Ketotestosterone, but not estradiol-17β, significantly increased oocyte diameters at physiological doses (30–100 ng/mL). Similarly, incubation with insulin-like growth factor-I, but not any of the other peptide hormones tested, resulted in increased oocyte diameters at high physiological doses (≥30 ng/mL). These data implicate androgens and hormones from the metabolic axis in mediating growth of previtellogenic oocytes in eel. 相似文献
16.
ABSTRACT: Final oocyte maturation and ovulation of captive chub mackerel Scomber japonicus with fully yolk-accumulated oocytes were induced by a single injection of human chorionic gonadotropin. Reproductive parameters, including spawning frequency and batch fecundity, which are required to estimate spawning biomass in pelagic fish by the daily egg production method, were analyzed. Germinal vesicle migration (GVM) occurred at 18–24 h post-injection, and the hydration and ovulation of oocytes were completed at 30 and 36 h post-injection, respectively. The results of the maturation process suggest that fish with GVM-stage ovaries captured in the daytime from the field are capable of spawning on the night following their capture. The oocytes used in the oocyte size-frequency distribution method for batch fecundity estimates should be at late GVM and more advanced stages. The results of sequential artificial insemination showed that the quality of ovulated eggs held in the ovarian lumen rapidly deteriorated as time progressed after ovulation. This indicates that the fertilization window for the ovulated eggs of chub mackerel lasts only a few hours, and spawning behavior should be performed within a few hours after ovulation in the wild population. 相似文献
17.
Thermal shock can disrupt meiosis II in fish oocytes, producing embryos with an extra set of maternal chromosomes from the second polar body; such shocks have been used to induce diploid gynogenesis or triploidy with varied success in different salmonid species. We investigated the effect of thermal shocks on the success of inducing diploid gynogenesis in chinook salmon (Oncorhynchus tshawytscha) oocytes. Different times of initiation (8, 16, and 24 min post-activation) resulted in similar survival. A treatment duration of 20 min resulted in significantly higher gynogen survival to hatch compared to 5 or 15 min, and a 10-min duration resulted in the lowest mean survival. The effect of temperature on induced retention of the second polar body is unclear due to varied results dependent on the maternal egg source; however, the mean survival to hatch for 25°C treatment groups was significantly higher than the mean of 29°C groups. 相似文献