共查询到19条相似文献,搜索用时 93 毫秒
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以小鼠为动物模型,研究延长小鼠卵母细胞在含有冷冻保护剂的液体中的平衡时间,并且降低或提高平衡时的温度是否能影响卵母细胞完成减数分裂的能力。结果表明:1)无论是否有DMSO,0℃平衡15min或60min后卵母细胞的激活率没有明显差异;2)1.5mol/L DMSO液中,0℃平衡15min和60min后卵母细胞激活率低于对照组;3)0℃平衡15min和60min后卵母细胞的激活率与对照组没有差异;4)1.5mol/L的DMSO液体中24℃平衡15min和60min后卵母细胞的激活率显著低于对照组;5)无论是否添加DMSO,无论是24℃平衡还是0℃平衡均没有影响激活卵母细胞的第二极体排出,说明卵母细胞低温平衡时添加DMSO对卵母细胞的激活有一定的影响。 相似文献
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本研究探讨了磷酸二酯酶抑制剂米力农(milrinone)对水牛卵母细胞体外自发成熟和促性腺激素诱导成熟的影响,以便提高卵母细胞体外成熟质量。试验对水牛卵丘卵母细胞复合体(COCs)培养不同时间或采取不同处理后,取出卵母细胞剥光后固定,然后用间苯二酚蓝染色,观察卵母细胞核成熟情况。结果表明:(1)Milrinone对水牛COCs的自发成熟具有抑制作用,且具有剂量依赖关系;(2)Milrinone对水牛卵母细胞体外自发成熟的抑制作用随培养时间的延长没有减弱,可作为水牛卵母细胞成熟过程中的核成熟抑制剂;(3)Milrinone能显著抑制促卵泡激素(FSH)诱导的水牛COCs体外成熟,但是随着时间的延长,这种抑制作用可以部分被FSH克服。 相似文献
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探讨了生殖老化对小鼠卵母细胞后期发育及基因表达水平的影响。结果表明,与青年小鼠相比,老龄小鼠MII期卵母细胞经体外受精后早期胚胎原核形成的时间发生延迟(1 h),囊胚发育率有显著差异(61.98%vs.70.95%,P0.05)。提取老龄和青年CD1小鼠MII期卵母细胞总RNA,使用基因表达谱芯片对其进行分析。结果表明,生殖老化导致1 737个基因表达表现出显著差异,其中64.9%的基因表达上调,35.1%的基因表达下调,生殖老化导致Hdac10等基因产生极显著的表达差异,小鼠卵母细胞质量下降,最终影响其后期发育能力。 相似文献
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小鼠卵母细胞冷冻保存技术研究进展 总被引:1,自引:0,他引:1
毛寿林 《养殖与饲料.饲料世界》2008,(2):5-7
卵母细胞的冷冻保存是动物繁殖技术领域生殖细胞冷冻的研究热点。在此介绍了小鼠卵母细胞冷冻保存的研究进展,阐述了小鼠卵母细胞冷冻保存的原理,探讨了冷冻保护剂的作用机理。最后,对小鼠卵母细胞的冷冻保存方法及其原理进行了概括。 相似文献
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试验选用5-6周龄雌性昆明小鼠的成熟卵母细胞,在不同前处理液(10%EG或10%EG+10%DMSO)中平衡5min,然后在冷冻溶液(EFS30、EFS40、EDFS30或EDFS40)中平衡30s后进行OPS法和SSV法玻璃化冷冻保存。试验结果表明:(1)小鼠成熟卵母细胞的OPS法冷冻保存,用EFS冷冻的卵母细胞解冻后形态正常率为75.8%,激活后卵裂率为47.8%;在EDFS30中冷冻保存,解冻后形态正常率为87.2%,卵裂率为62.0%;在EDFS40中冷冻保存,解冻后形态正常率为86.6%,卵裂率为57.4%,与对照组(99.0%和80.5%)相比,差异极显著(P〈0.01)。(2)小鼠成熟卵母细胞的SSV法冷冻保存,用EFS冷冻的卵母细胞解冻后形态正常率为82.7%,卵裂率为47.1%;在EDFS30中冷冻保存,解冻后形态正常率为91.5%,卵裂率为57.5%;在EDFS40中冷冻保存,解冻后形态正常率最高为85.1%,卵裂率为55.9%,与对照组(99.0%和80.5%)相比,差异极显著(P〈0.01)。(3)OPS法和SSV法冷冻小鼠成熟卵母细胞解冻后形态正常率为86.5%和91.6%,卵裂率为53.8%和55.6%.与对照绢相比。差异极显著(P〈0.01)。 相似文献
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采用PBS作为冷冻基础液,分别用甘油和二甲基亚砜(DMSO)作为冷冻保护液,在程序化冷冻保存和玻璃化冷冻保存条件下,研究小鼠生发泡期(GV期)卵母细胞的抗冻能力。结果表明,2种冷冻方法对小鼠GV期卵母细胞解冻后形态正常率和存活率无显著影响(P>0.05)。冷冻保护剂种类对小鼠GV期卵母细胞解冻后形态正常率无显著影响(P>0.05);但对存活率有显著影响,玻璃化冷冻采用二甲基亚砜作为冷冻保护液效果极显著优于甘油(P<0.01)。以冷冻效果较好的二甲基亚砜作为冷冻保护液,采用玻璃化冷冻不同发育阶段(GV期和MⅡ期)的小鼠卵母细胞,解冻后形态正常率无显著差异(P>0.05),但存活率GV期要显著优于MⅡ期卵母细胞(P<0.05)。 相似文献
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为探讨不同激活剂的卵母细胞孤雌激活效果及激活胚体外发育情况,用乙醇、CaA23187、SrCl2、6-DMAP及CB分别对小鼠卵母细胞进行了激活处理.结果显示,70 mL/L乙醇刺激小鼠卵母细胞时,以激活5~7 min的激活效果及孤雌胚发育较好,桑囊胚发育率可达29.11%;用SrCl2激活小鼠卵母细胞时,6~10 mmol/L为最佳处理浓度,3~6 h为最佳激活时间,桑囊胚发育率可达16.67%;以70 mL/L乙醇激活5 min,再用2 mmol/L 6-DMAP+5 μg/mL CB激活3 h效果最佳,桑囊胚发育率高达35.77%.研究证实,小鼠卵母细胞经乙醇、6-DMAP和CB等复合激活后能较好地发育. 相似文献
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The present study examines the contribution of the nucleus to meiotic competence in mouse oocytes that were reconstructed using nuclear transfer. Three types of reconstructed oocytes were produced: MP‐GV, by transplanting the male pronucleus (MP) into germinal vesicle (GV) stage oocytes; 3T3‐GV, by transplanting the nucleus of a National Institute of Health (NIH) 3T3 cell into a GV stage oocyte; and 3T3‐MII, by transplanting the nucleus of an NIH 3T3 cell into a metaphase II (MII) stage oocyte. The fusion rates differed, but not significantly, in the MP‐GV, 3T3‐GV, and 3T3‐MII groups (77, 63, 56%, respectively). Then, meiotic competence was compared in MP‐GV, 3T3‐GV and non‐manipulated GV stage oocytes as a control. Nuclear envelope breakdown occurred in all the reconstructed oocytes, as well as the control ones. The percentage of first polar body extrusion differed between the MP‐GV (100%), 3T3‐GV (72%), and control (67%) groups. DNA staining with Hoechst 33342 revealed that in the MP‐GV‐group oocytes that had reached MII stage, the chromosomes were condensed and aligned in a regular array similar to the normal metaphase plate. By contrast, in 3T3‐GV group oocytes, the condensed chromosomes were irregularly scattered in the cytoplasm. These results suggest that the donor nucleus affects meiotic competence in reconstructed oocytes. 相似文献
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Changes in the cytoplasmic inclusions during meiotic maturation were histochemically examined in cultured porcine oocytes. The oocytes contained a small amount of protein and glycogen granules throughout the maturation culture, as well as Sudanophilic lipids composed of small, medium and large droplets. Soon after collection, the amount of Sudanophilic lipid droplets of small and medium size was small and there were 167 ± 11.2 large droplets. After being cultured for 22 h, the number of large lipid droplets decreased remarkably, while the number of small and medium ones increased. There were no differences in the number of Sudanophilic lipid droplets of different sizes between ovulated oocytes and the oocytes cultured for 44 h. The oocytes always contained a large amount of neutral fats and lipoids, but not cholesterols. In the oocytes cultured for 22 h with olomoucine, both the resumption of nuclear maturation and the decrease in the size of the Sudanophilic lipid droplets were inhibited. From the present findings, it appears that the change in the size of the Sudanophilic lipid droplets in the cytoplasm of porcine oocytes is closely related to nuclear maturation. 相似文献
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Senyang Cao Shaoping Huang Ying Guo Lin Zhou Ying Lu Shanshan Lai 《Reproduction in domestic animals》2020,55(11):1607-1618
Oocyte proteins play an important role in oocyte maturation, fertilization and embryonic development. However, the protein composition of mouse germinal vesicle (GV) oocytes is still unclear. Using one-dimensional Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (1D SDS-PAGE) and Reverse-phase liquid chromatography tandem mass spectrometry (RP-LC-MS/MS), we constructed a protein profile of mouse GV oocytes. First, our proteomics profile identified 1,405 different proteins from 11,000 mouse GV oocytes lacking zona pellucida. Second, with detailed bioinformatics analysis, a group of proteins that play an essential role in oocyte maturation was screened. In addition, the expression and localization of suppressor of G2 allele of skp1(SUGT1, also called SGT1), heterogeneous nuclear ribonucleoprotein K (Hnrpk), Seruin, Cullin1(Clu1) and nuclear distribution protein C (Nudc) in mouse ovaries and early embryos were also captured and investigated in this study. Moreover, the protein profile was submitted to the Proteomics Identifications Database (PRIDE) and is available via ProteomeXchange with the identifier PXD014314. Our research provides valuable resources for the study of oocyte proteins and oocyte maturation and helps to clarify the mechanisms of oocyte maturation. 相似文献
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Yuanyuan He Yile Wang Hengde Zhang Yong Zhang Fusheng Quan 《Reproduction in domestic animals》2021,56(4):545-554
Oxidative stress inevitably occurs during oocyte maturation in vitro. α-lipoic acid (α-LA) has a strong antioxidant capacity, but the effect of α-LA on parthenogenetic activation of oocytes was rarely reported. This study aims to investigate the effect of supplementing α-LA to in vitro maturation medium on the subsequent developmental ability of goat parthenogenetic embryos during oocytes maturation. In the study, the goat cumulus-oocyte complex was divided into the experimental (with 25 μmol/L α-LA) and the control (without α-LA) groups. Oxidase expression was measured using RT-qPCR. After 18–22 hr of maturation, the oocytes were then parthenogenetic activated. The total antioxidant capacity of embryos was measured after 0, 24, 48, 72 and 96 hr of culture. Rates of oocyte maturation and the rates of development for parthenogenetic embryos in the α-LA group were significantly improved by 7.88% (p < .05) and 5.41% (p < .05) compared with those in the control group, respectively. After 24 hr, the difference in total antioxidant capacity was extremely significant in both groups. An evident decrease in the control group and a minor decrease in the α-LA group were observed (p < .01). The ratio of inner cell mass cells to the total cell number of blastocysts in the α-LA group increased compared with that in the control group (p < .05) on day 8. α-LA significantly promoted the expression of SOD and GPX4 of parthenogenetic blastocysts and maturated oocytes. α-LA (25 μmol/L) improved the maturation rate and the developmental competence of the parthenogenetic activation of oocytes, which might be mediated by maintaining the total antioxidant ability of oocytes during the culture period. 相似文献
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Jung-Taek Kang Dae-Kee Kwon Sol-Ji Park Su-Jin Kim Joon-Ho Moon Ok-Jae Koo Goo Jang Byeong-Chun Lee 《Journal of veterinary science (Suw?n-si, Korea)》2013,14(1):15-20
Quercetin is a plant-derived flavonoid found in fruits or vegetables that has antioxidant properties and acts as a free radical scavenger. We investigated the effects of quercetin on porcine oocyte nuclear maturation and embryonic development after parthenogenetic activation. We then evaluated the antioxidant activities of quercetin by measuring reactive oxygen species (ROS) levels in matured oocytes. Immature oocytes were untreated or treated with 1, 10, and 50 µg/mL quercetin during in vitro maturation (IVM). Quercetin treatment did not improve oocyte nuclear maturation, but significantly higher blastocyst rates (p < 0.05) of parthenogenetically activated oocytes were achieved when the IVM medium was supplemented with an adequate concentration of quercetin (1 µg/mL). However, cleavage rates and blastocyst cell numbers were not affected. Oocytes treated with 1 or 10 µg/mL quercetin had significantly lower (p < 0.05) levels of ROS than the control and group treated with the highest concentration of quercetin (50 µg/mL). Moreover, this highest concentration was detrimental to oocyte nuclear maturation and blastocyst formation. Based on our findings, we concluded that exogenous quercetin reduces ROS levels during oocyte maturation and is beneficial for subsequent embryo development. 相似文献