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试验采用不同浓度的DTT或GSH预处理猪精子,统计ICSI后精核解聚率、雄原核形成率及胚胎发育情况.结果表明,①精核解聚率随DTT浓度的增加而提高,6 mmol/L DTT处理组的2PN 2Pb形成率(P＜0.01)和卵裂率(P＜0.05)均高于其余各组.随着DTT浓度的增加,囊胚总细胞数呈现下降趋势.表明6 mmol/LDTT预处理精子的ICSI效果较好,但DTT在促进精核解聚的同时所产生的副作用在一定程度上影响了ICSI胚胎的后期发育;②随着GSH浓度的升高,注射卵母细胞的2PN 2Pb形成率、MPN形成率和精核解聚率均呈上升趋势.0.5 mmol/L GSH处理组的卵裂率高于其余处理组(P＜0.05)和对照组(P＜0.01),囊胚发育率显著高于对照组(P＜0.05).说明0.50 mmol/L GSH处理组的ICSI效果较好;③0.5 mmol/L GSH处理组的2PN 2Pb形成率、卵裂率和囊胚总细胞数极显著高于6 mmol/LDTT处理组(P＜0.01),两者MPN形成率差异显著(P＜0.05).表明0.5 mmol/L GSH预处理精子的ICSI效果要好于6 mmol/L DTT. 相似文献
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精子不同处理和胰岛素不同浓度对猪卵母细胞胞质内单精子显微受精的影响 总被引：1，自引：0，他引：1
探讨了分别使用新鲜、冷冻和超声波断尾精子以及在胚胎培养液中分别添加不同浓度胰岛素对猪卵母细胞胞质内单精子显微受精(ICSI)胚胎早期发育的影响.结果:(1)使用冷冻解冻精子与新鲜精子相比对猪卵母细胞ICSI后的卵裂率和囊胚率均无显著影响(P＞0.05);2)精子断尾与否对猪卵母细胞ICSI后的分裂率和囊胚率没有显著影响(P＞0.05);3)在胚胎培养液中添加5 mg/L胰岛素与对照组相比可显著提高猪ICSI胚胎的囊胚发育率(18.22% vs 3.60%,P＜0.05). 相似文献
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实验探讨了精卵因素对母牛卵母细胞质内精子显微注射效果的影响.结果表明:新鲜精液组和性控冻精组ICSI对卵母细胞存活率、卵裂率和囊胚率均无显著差异(P>0.05);采用A、B、C级精子穿刺Ⅰ、Ⅱ、Ⅲ级卵母细胞时,穿刺卵受精率、卵裂率和囊胚率在三者之间无显著差异(P>0.05),总体效果A>B>C、Ⅰ>Ⅱ>Ⅲ;D级精子穿刺Ⅱ级卵母细胞后的穿刺卵受精率显著低于Ⅰ级卵母细胞(P<0.05),卵裂率和囊胚率与Ⅰ级卵母细胞相比无显著差异(P>0.05),穿刺Ⅲ级卵母细胞后的穿刺受精率和囊胚率均显著低于Ⅰ级卵母细胞(P<0.05),受精卵的卵裂率与Ⅰ级卵母细胞相比则无显著差异(P>0.05).由此可见:采用流式细胞仪分离的性控冻精与新鲜精液相比,对ICSI后的受精率和囊胚率无显著影响;精子活动力和卵子质量对ICSI的效果均有一定程度影响,且随精子活动力的降低和卵母细胞质量的下降而逐渐显著. 相似文献
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本研究旨在探讨不同的联合激活处理、精子预处理方法和单精子注射方式对奶牛分离精子ICSI效率的影响。借助显微操作仪将经不同预处理的奶牛分离X精子直接注入体外成熟22～24h的牛卵母细胞胞质内,注射过程采用回吸胞质和不回吸胞质2种处理,最后分别用3种激活方案对注射卵进行激活。结果表明:用CR1aa+Ionomycin+6-DMAP对ICSI注射卵进行激活后,卵裂率和囊胚率都高于A23187+CHX和7%ET+CHX两种联合激活组(63.74%vs61.09%、53.75%;28.54%vs17.68%、22.00%,P0.05);采用percoll密度梯度离心法处理精子,卵裂率(77.10%vs62.19%,P0.05)与囊胚率(26.95%vs22.82%,P0.05)均高于上游法处理组;ICSI时回吸胞质获得的注射卵,其卵裂率和囊胚率均显著高于不回吸胞质处理组(63.10%,25.35%vs41.56%,19.40%,P0.05)。结果表明,用percoll密度梯度离心法处理精子、注射时回吸胞质、用CR1aa+Iono-mycin+6-DMAP激活ICSI注射卵,可显著提高奶牛分离精子ICSI的效率。 相似文献
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本试验探讨了不同辅助激活方法(Calciumionophore A23187激活、Calciumionophore A23187+6-DMAP联合激活和电激活)、不同精子预处理方法(液氮冻融处理和0.1%Triton X-100处理)和在添加半胱氨酸的胚胎培养液中培养不同时间(0 h、4 h、12 h和168 h)对猪卵母细胞内单精子注射(ICSI)胚胎体外发育的影响。结果显示:与无辅助激活相比,A23187+6-DMAP联合激活和电激活均能显著提高ICSI卵母细胞的激活率、卵裂率和囊胚率(P0.05),A23187+6-DMAP联合激活能显著提高ICSI卵母细胞的受精率(P0.05)。液氮冻融精子组ICSI卵母细胞的雄原核形成率显著高于活精子组(P0.05)。在添加半胱氨酸的胚胎培养液中培养4 h的ICSI卵母细胞受精率、雄原核形成率和囊胚率显著高于0 h组(P0.05)。以上结果表明,猪卵母细胞在ICSI后需要辅助激活来启动胚胎顺利发育,A23187+6-DMAP激活效果较好。液氮冻融精子可以促进ICSI后雄原核的形成。半胱氨酸处理4 h对猪ICSI卵母细胞受精和发育均有促进作用。 相似文献
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本文对影响绵羊精子胞质内显微受精的因素进行分析。结果如下:1、操作液中PVP浓度分别为8%和5%时,卵裂率为(83.6±2.98)%和(72.0±2.53)%,囊胚发育率为(30.5±1.95)%和(16.0±1.33)%,均差异显著(P0.05)。2、操作液中添加CB和不添加CB,卵裂率为(77.5±3.17)%和(74.7±2.09)%,没有显著影响(P0.05),但添加CB的囊胚发育率(29.2±2.81)%却显著高于对照组(14.3±1.99)%(P0.05)。3、在培养液(IVC)中添加VEGF和不添加VEGF,卵裂率为(78.7±3.34)%和(83.9±1.95)%;囊胚发育率为(27.7±2.76)%和(23.6±1.29)%,差异不显著(P0.05)。4、在ICSI过程中恒温和室温条件下,卵裂率为(80.1±3.18)%和(90.6±3.02)%,差异显著(P0.05),囊胚发育率为(31.3±3.87)%和(28.2±3.34)%,没有显著影响(P0.05)。5、在4 h以前注射卵裂率分别为(84.7±3.62)%、(86.0±2.89)%、(80.6±3.04)%,没有显著差异(P0.05)。0～1 h注射的囊胚发育率(33.6±3.37)%,要显著高于1～2 h注射的(P0.05),并极显著高于3～4 h的(P0.01)。1～2 h与3～4 h注射的囊胚发育率分别为(18.3±2.52)%和(13.0±2.04)%,没有明显差异(P0.05)。 相似文献
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奶牛体外受精效果几种影响因素的研究 总被引：1，自引：0，他引：1
文章研究了卵巢的保存温度、保存时间、不同的培养液成分、培养方法等对奶牛体外受精后的卵裂率、囊胚发育率的影响.结果表明①采集的卵巢在37℃保存2h时,卵巢卵母细胞体外受精后的卵裂率(71.3%)和囊胚率(31.4%),与25℃保存组的卵裂率(75%)和囊胚率(30.7%)相比,差异不显著(P＞0.05);但与40℃保存组(55%和15%)和4℃保存组(47.8%和12%)有显著差异(P＜0.05);当卵巢保存6h时,25℃保存组的卵裂率(70.2%)和囊胚率(29.2%)均显著高于37℃保存组(50.9%和13%)(P＜0.05);与40℃保存组(30%和7%)和4℃组(30.1%和8.2%)差异极显著(P＜0.01);但与保存2 h组差异不显著.在25℃温度下,屠宰牛卵巢保存时间达到8h时,卵母细胞的卵裂率和囊胚率显著下降,分别为55%和14%.②作为早期胚胎的培养液,CRlaa的效果好于TCM-199.③50μl微滴组的卵裂率(71%)与微滴100μl组(75%)和500μl组(72%)无显著差异(P＞0.05),但囊胚率(15.8%)显著低于100μl组(32%)(P＜0.05);大体积培养组(2 ml)的卵裂率和囊胚率(55%和14.8%)显著低于100μl组和500μl组(P＜0.05). 相似文献
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猪体外受精(in vitro fertilization,IVF)技术效率的不稳定性限制了该技术在畜牧生产中的应用。本研究比较了精子获能前后的状态和活精子数之间的关系;精子获能后的浓度和状态对IVF后胚胎卵裂率、囊胚率的影响;不同个体来源的精子混合后对IVF卵裂率、囊胚率的影响。结果表明:精子获能前后,活精子数无显著性差异(P>0.05);检测精子的浓度与状态与卵裂率、囊胚率无直接关联;利用混合精子进行IVF,单一精子所得到的IVF囊胚率分别是A17.00%、B 12.24%、C 33.00%、D 4.81%、E 10.00%、F 22.00%,混合精子的囊胚率分别是A+B 77.00%、C+D 67.01%、E+F 35.50%,混合精子和单一精子的囊胚率差异不显著。 相似文献
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牦牛卵母细胞的体外成熟、种间受精与胚胎培养 总被引：1，自引：0，他引：1
探讨了卵母细胞体外成熟时间、卵母细胞质量、精子准备和受精卵培养体系对牦牛卵母细胞种间体外受精效果的影响。结果表明:牦牛卵母细胞随体外成熟培养时间延长,第一极体排出率增加,囊胚发育率以成熟培养24 h最高;A级卵母细胞种间受精后囊胚的发育率(48.77±3.76)%显著高于B级(32.05±5.24)%和C级(7.54±7.18)%(P<0.05);BO液洗涤离心处理的精子受精后卵裂率(82.53±6.54)%显著高于Percoll液分离精子受精后的卵裂率(67.39±4.50)%(P<0.05),而两者囊胚率差异不显著((42.32±4.13)%vs(35.59±5.62)%,P>0.05);荷斯坦奶牛精子浓度在1×106～5×106/mL范围与牦牛卵母细胞受精效果差异不显著。卵丘细胞、输卵管上皮细胞共培养和SOF液培养种间受精卵,其卵裂率差异不显著,但共培养组的桑葚胚、囊胚和孵化胚发育率均显著高于SOF液培养组。 相似文献
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Sperm binding and sperm penetration of the zona pellucida (zp) are regarded as species‐specific. In this investigation, the interactions between bovine oocytes and porcine, respectively, equine spermatozoa have been studied under in vitro conditions and compared with the normal in vitro fertilization of bovine oocytes by bovine sperm. Surprisingly, many of the heterologous spermatozoa adhered firmly to the bovine oocytes and could not be removed by intense washing. On average, more than 100 boar or equine spermatozoa were bound to the zp of bovine oocytes. Electron microscopic studies clearly demonstrated that porcine sperm attached to the zona and underwent the acrosome reaction. Equine spermatozoa displayed a similar binding affinity, but unlike the porcine spermatozoa even penetrated the zp and were taken up into the oocyte after a longer period of co‐incubation. Considering these new results the dogma of a strict species specificity of sperm zona interactions under in vitro conditions has to be reconsidered. 相似文献
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JF Asturiano F Marco-Jiménez DS Peñaranda DL Garzón L Pérez JS Vicente M Jover 《Reproduction in domestic animals》2007,42(2):162-166
The main objective of the present work was to study the effect of cryopreservation of European eel sperm both on the sperm viability and the spermatozoa head morphology. Spermatozoa morphology was evaluated with computer-assisted morphology analysis after collection in fresh samples, after adding the freezing medium containing dimethyl sulfoxide as cryoprotectant and, finally, after the cryopreservation process and thawing. Cell viability was assessed, in both fresh and thawed samples, by Hoechst 33258 staining. Computer-assisted sperm analysis (CASA) was used to determine the percentage of motile cells and to measure motility parameters in sperm samples. A significant decrease of head perimeter (12.56%) and area (17.90%) was detected from spermatozoa in fresh to thawed samples, indicating that cells do not recover the original size after the cryopreservation process. CASA was used to measure the percentage of motile cells (51.9%) and spermatozoa motility parameters such as curvilinear, straight line and angular path velocities, as well as beating cross frequency. This technique was employed in the fresh sperm samples but proteins present at the freezing medium (L-alpha-phosphatidylcholine) made impossible to use this last technique in thawed samples. When sperm viability was assessed by Hoechst staining, a significant decrease of approximately 15% (73.10 vs 58.26%) of alive spermatozoa was registered from fresh to thawed samples. The percentage of motile cells measured by CASA in fresh samples (51.9%) was lower than the percentage of alive cells determined by Hoechst stainning, suggesting the existence of different batches of spermatozoa in different stages of development, even during the eight to tenth weeks of treatment, when the highest sperm quality was found. 相似文献
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The present study investigated daily sperm output and sperm morphology of fresh semen in eight Swedish Warmblood stallions aged 5–8 years. They were used for artificial insemination, and their fertility during the breeding season of semen collection exceeded 60% per cycle. One ejaculate of semen was collected daily for 10 consecutive days from each stallion. The gel-free volume was measured, and the sperm concentration was assessed with a Bürker chamber. The volume of gel-free fraction was multiplied by the sperm concentration to give the total number of spermatozoa (TSN). Sperm morphology was examined in ejaculates collected on days 2, 5 and 10. An aliquot from each ejaculate was fixed in 1 ml formol–saline immediately after collection and examined under a phase-contrast microscope (magnification 1000×) to assess morphological abnormalities. Furthermore smears were prepared and stained according to Williams (carbolfuchsin–eosin) for a more detailed examination of the sperm heads under a light microscope (magnification 1000×). Analysis of variance was applied to data. Total spermatozoa number decreased progressively during the first 8 days of collection, and daily sperm output (DSO) was calculated as mean TSN of collections on days 8–10, being 6.4 × 109 spermatozoa. The overall percentages of morphologically normal spermatozoa in ejaculates collected on days 2, 5 and 10 were above 70%, being significantly lower in ejaculate 2 (68.6%) compared with ejaculates 5 and 10 (72.9% respectively 75.3%). 相似文献
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利用液体闪烁计数仪测定绵羊精子的超弱化学发光,以研究它的机理和畜牧中的应用价值。结果表明,精子的发光与活力、呼吸、果糖分解、~(32)Pi摄入量和磷酸肌酸呈正相关(r=0.9806,P<0.01;0.9684,p<0.01;0.9882,p<0.01;0.9793,p<0.01;0.9962,p<0.01)说明了精子的发光与代谢存在有内在的联系,反映了精子的能量转化过程和精子细胞物理化学反应的信息,是评定精子质量很有价值的一个指标。 相似文献
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精子分离技术的研究进展 总被引：2，自引：0，他引：2
控制家畜性别一直是人们所感兴趣的话题 ,通过性别控制可以大幅度提高生产力 ,为人类提供更多更好的畜产品。分离Ｘ精子和Ｙ精子是最简单、最经济的性别控制途径 ,这种方法主要依据Ｘ精子和Ｙ精子不同的物理特性 (体积、密度、电荷、运动性 )和化学特性 (ＤＮＡ含量、特异性抗原 )进行分离。1 性别控制技术简介1 1 性别控制的意义动物性别控制技术是通过对动物的正常生殖过程进行人为干预 ,使雌性动物产出人们期望性别后代的一门生物技术。家畜性别控制在畜牧生产中有着重要的意义。首先 ,可以充分发挥受性别限制的生产现状 (如泌乳 )和受… 相似文献
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大熊猫是我国特产的濒危珍稀动物,由于在野外的种群数量的大量减少,人们正试图通过圈养大熊猫的途径来挽救大熊猫。但圈养大熊猫的雄兽只有10%能进行自然交配,不能与公兽进行自然交配的雌兽只能进行人工授精,人工授精的成功率往往由于冻精的活力(Motility)、活率(Percentalive) 相似文献
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《Journal of Equine Veterinary Science》1996,16(9):390-392
A group of fertile mares was bred on the day before ovulation. Uterine flushes were collected from mares 48 h after breeding. Examination of stained cytocentrifuge preparations of the flushes revealed the presence of sperm in 4 of 10 flushes. The sperm were seen both intact and fragmented, outside and within neutrophils. Three of these mares became pregnant and therefore we conclude that sperm can still normally be present in the uteri of fertile mares 48h after breeding. 相似文献