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猪精液冷冻保存技术对生猪产业发展有重要意义,有利于提高优秀种公猪的利用率以及实现优秀种质资源的长久保存,促进种猪场降本增效。目前中国猪精液冷冻保存技术取得了一系列进步,国内应用猪冻精配种的受胎率以及产活仔数基本达到了国外冻精配种水平。但是,由于冻融过程会对猪精子造成一定程度的损伤,精子的受精能力下降,受精窗口期缩短,国内、外应用冻精配种的生产成绩仍与常温保存精液存在不小差距。文章回顾了国内、外猪精液冷冻保存技术的发展历程,介绍了猪精液和精子的基本特点,简述了降温过程造成精子损伤的机理,并从冷冻稀释液成分这一角度分析介绍了减小冷冻损伤的机理与方法,如添加冷冻保护剂减少细胞内外冰晶形成、维持细胞内外渗透压以及稳定细胞膜结构;添加抗菌物质抑制解冻后细菌的快速繁殖;添加抗氧化物降低活性氧水平,缓解氧化应激,保护精子结构和受精能力等,最后对猪精液冷冻保存技术未来的发展方向进行展望,以期为相关研究以及实际应用提供参考。 相似文献
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牛精液冷冻与人工授精技术相结合，在牛品种改良、保护优良种质资源中发挥着重要作用。尽管冷冻后部分牛精子活力高达60%以上，但是冷冻后精子受损现象却普遍存在，尤其是受精能力与鲜精相比显著降低。作者详细介绍了冷冻-解冻过程对牛精子造成的主要损伤，包括精子的形态完整性、活力及遗传物质的改变等；阐述了冷冻造成精子损伤的主要原因，即细胞内冰晶形成和氧化应激反应及其产生的可能机制；并且对目前常用的提高冷冻精子质量的方法，如添加冷冻保护剂、优化冷冻程序以及添加抗氧化剂等进行了详细地综述；提出了在冷冻精液研究方面值得探索的问题，以期为家畜精液冷冻保存技术的进一步优化提供理论依据。 相似文献
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《国外畜牧学(猪与禽)》2021,41(3)
猪精液冷冻后会导致精子活力不足,从而使猪冷冻精液在生产中的应用效果不理想,未得到大范围的使用,本文对甘肃省猪冷冻精液技术进行了分析,以促进猪冷冻精液技术的研究。 相似文献
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精液冷冻技术在驴种质资源的长期保存和引进以及人工辅助繁殖技术应用等方面具有重要意义,精液冷冻保存有利于充分发挥优秀种公驴的繁殖力,降低驴养殖产业的生产成本。近年来,驴精液冷冻保护剂和冷冻方法不断优化,驴精液冷冻保存技术也取得了很大的进步。但由于驴冷冻复苏的精子活力和受胎率低,严重限制了驴冷冻精液的推广与应用。本文对驴精液冷冻保存技术的发展历程、精子冷冻损伤机制、影响驴精液冷冻保存的因素和驴冷冻精液的应用进行综述,为今后驴精液冷冻保存技术的研究提供参考。 相似文献
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为优化猪精子载体法技术参数,利用荧光定量PCR、荧光显微镜检测和精液常规检测方法,分析不同DNA转染剂、不同孵育温度和不同形态DNA对猪精子转染外源DNA的影响。结果表明,PEI和TransFast转染剂能极显著提高猪精子转染效果,且PEI优于TransFast,而NanoFect转染剂包裹DNA不能转染精子。随着转染温度的升高,精子的活力和活率均明显下降,而转染率和内化外源DNA量基本保持稳定,而吸附外源DNA量呈下降趋势。环状DNA和线性化DNA在与精子共孵育后,两者的精子活力、活率、转染率和内化外源DNA量差异均不显著,精子吸附线性化DNA量极显著高于环状DNA量。综合分析表明,外源DNA形态对猪精子转染效果无显著影响;PEI和TransFast能够显著提高猪精子转染效果;共孵育温度以17℃为最佳,本结果为开展精子载体法制备转基因猪研究提供了基础试验依据。 相似文献
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近年来，随着畜牧业的发展，家畜精液保存技术在生产实践中发挥着重要作用。在精液保存过程中，体内外环境的差异打破了精子所处环境的动态平衡，保存期间产生大量的活性氧（reactive oxygen species，ROS）。过量的ROS会与精子质膜发生氧化反应，破坏精子的功能和结构完整性，使其活力和质量降低，影响受胎率。为了降低ROS对精液的损伤，加入外源抗氧化剂可以有效抑制和清除ROS，避免精子细胞受到破坏，进而提高精液品质。文章对抗氧化剂的作用机制、抗氧化剂的种类以及在猪精液保存上的应用等方面进行综述。 相似文献
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《畜牧与生物技术杂志(英文版)》2020,(1)
Background: The application of cryopreservation and artificial insemination technology have contributed to the advancement of animal reproduction. However, a substantial proportion of spermatozoa undergoes alterations and loses their fertility during cryopreservation, rendering the frozen-thawed semen impractical for routine use.Cryopreservation is known to reduce sperm lifespan and fertility. Variation in cryosurvival of spermatozoa from different sires and even with the individual sire is common in artificial insemination(AI) centers. Our goal is to improve post-thawed semen quality by optimization of cryopreservation technique through sperm selection prior to cryopreservation process.Results: Our strategy of sperm selection based on rheotaxis and thermotaxis(SSRT) on macrosale in a rotating fluid flow demonstrated the ability to maintain the original pre-freezing structural integrity, viability and biological function related to fertilization competence. This strategy has a positive effect on the cryosurvival and fertilizing abilities of spermatozoa as supported by the improvement on pregnancy rate of Japanese Black heifers and Holstein repeat breeders. This technique protected further sublethal damage to bovine spermatozoa(higher %cryosurvival than the control) and resulted in the improvement of DNA integrity. Prefreeze selected spermatozoa demonstrated slower and controlled capacitation than unprocessed control which is thought to be related to sperm longevity and consequently to appropriate timing during in vivo fertilization.Conclusions: These results provide solid evidence that improvement of post-thawed semen quality by SSRT method is beneficial in terms of cryosurvival, longevity of post-thawed sperm, and optimization of in vivo fertilization, embryo development and calving as supported by the favorable results of field fertility study. 相似文献
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实验利用透射电镜技术和扫描电镜技术对冷冻/解冻后的塔里木马鹿精子的超微结构进行了观察。结果表明:电镜下观察,塔里木马鹿精子头部呈扁平的卵圆形,长约5.95μm。颈部很短且不明显,长约0.45μm,宽为0.62μm。尾部中段横切面近圆形,直径约为0.58μm,轴丝为9×2+2型;线粒体鞘螺旋段为64～70转。尾部末段仅见质膜包围,9束微管和1对中央微管,形成9+2微管结构。冷冻/解冻后塔里木马鹿部分精子结构发生了变化,一些精子肿胀、质膜皱褶、扭曲,部分质膜损坏或溃散;顶体轻度肿胀,顶体外膜凸凹不平,形成突起。冷冻对头部破坏程度较小。 相似文献
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Cryopreservation of boar spermatozoa offers an effective means of long‐term storage of important genetic material. Many researchers have investigated how to improve reproductive performance by artificial insemination (AI) using cryopreserved boar spermatozoa. Recently, we and other groups reported that high conception rates (70–80%) can be achieved by AI with frozen‐thawed boar spermatozoa using a modified temperature program during freezing, or a novel cryopreservation extender to improve sperm quality (including sperm survivability, motility, membrane status and fertilization ability) after thawing, or a novel sperm infusion method, deep intra uterine insemination. However, these techniques have not yet been used for commercial pig production. The variation in sperm freezability among boars or among ejaculations in an identical boar is one of the main reasons for this problem. In our previous study, it was revealed that some components of seminal plasma have a negative effect on the freezability of boar sperm. One of these factors is bacteria‐released endotoxin (lipopolysaccharide: LPS). LPS binds to Toll‐like receptor‐4 (TLR‐4) expressed on the sperm surface, resulting in induction of apoptosis. On the other hand, seminal plasma suppresses cryo‐capacitation induced by thawing stress. On the basis of these findings, we designed a novel protocol of AI using frozen‐thawed boar sperm. 相似文献
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气象因素对种公牛精子发生的影响分析 总被引：4，自引：1，他引：3
本文采用相关和通径分析方法，分析了五项气象因子对精液量和精子活力的影响。结果显示，上两个月的月均最低温度（Ｘ４）和月均日照时数（Ｘ１）对精液量分别产生极显著（Ｐ〈０．０１）和显著（Ｐ〈０．０５）影响，当月平均温度（Ｘ２）和最高温度（Ｘ５）对精子活力产生显著（Ｐ〈０．０５）影响，降水量（Ｘ３）主要通过气温产生影响，五项气象因子对精子发生的总决定系数约３０％。这些结果对研究种公牛环境控制、精子发生有重 相似文献
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We previously identified 62, 39, 27 and 7 kDa porcine sperm plasma membrane proteins that demonstrated a predominant affinity for the porcine oocyte plasma membrane by Western ligand blotting. The current experiments were designed to further investigate the potential roles of these molecules in sperm–oocyte plasma membrane interaction. Abilities of these proteins to bind to the oocyte plasma membrane and to inhibit sperm–oocyte interaction were evaluated. Plasma membrane was isolated primarily from the head of ejaculated porcine sperm by nitrogen cavitation and density gradient centrifugation. Fractions containing the 62, 39, 27 and 7 kDa proteins were electroeluted from one dimensional SDS polyacrylamide gels, dialysed and proteins biotinylated. Following incubation with zona‐free porcine oocytes, bound protein was visualized with 20 μg TRITC‐avidin/ml using confocal microscopy. Fractions of the dialysed, electroeluted proteins were added to porcine in vitro fertilization assays. The 62, 39, 27 and 7 kDa proteins all demonstrated binding to the oocyte plasma membrane in contrast to a biotinylated control protein. Addition of unlabelled sperm plasma membrane proteins to the biotinylated protein visibly reduced binding. Addition of each of these protein fractions to in vitro fertilization assays reduced sperm interaction with the porcine oocyte plasma membrane in a concentration‐dependent manner. Binding of these sperm plasma membrane proteins to the oocyte plasma membrane and inhibition of fertilization are consistent with these proteins being involved in sperm–oocyte plasma membrane interaction. 相似文献
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单精注射法生产转GFP基因猪胚胎的研究 总被引：1，自引：1，他引：1
卵胞质内单精子注射(ICSI)的出现为治疗人类男性不育提供了新的途径。作为一种家畜胚胎工程新技术，ICSI可以解决猪的多精子受精问题。本研究用冷冻/解冻的猪精子与GFP基因孵育后对猪IVM卵母细胞进行ICSI，通过对猪ICSI卵母细胞发育能力和基因表达效率的分析，初步研究以精子为载体的转基因与卵细胞质内单精注射相结合的技术(SMGT-ICSI)生产转基因猪胚胎的技术路线的可行性。用GFP基因转染的精子注射后化学激活的ICSI卵母细胞基因表达率显著高于电激活处理的ICSI卵母细胞的基因表达率。用精子头部注射的猪卵母细胞和用完整精子注射的猪卵母细胞总基因表达效率无显著差异(P>0.05)。结果表明，用冷冻/解冻后的猪精子或精子头部与外源DNA孵育后通过ICSI方法生产转基因胚胎是可行的。 相似文献
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本实验以猪体外成熟卵子以及冷冻解冻后失活的精子为材料,以无BSA且成分明确的TL-HEPES溶液为操作液,探讨显微注射过程中分别使用钝口针和磨口针进行注射,对猪卵子的激活、ICSI胚胎的发育及EGFP表达效率的影响。结果表明:成熟后的猪卵子在不进行精子注射和电激活的情况下,使用钝口针空注射后,卵裂率显著高于磨口针(P<0.05)。分别使用钝口针和磨口针对成熟后的猪卵子进行精子注射,不论进行或不进行电激活,使用钝口针注射,卵子的卵裂率、囊胚率和胚胎EGFP的表达效率均显著高于磨口针(P<0.05)。本实验研究发现,使用钝口针进行注射有利于猪ICSI卵子的激活,胚胎的发育以及胚胎中EGFP表达效率。 相似文献