附睾中非编码小RNA(sncRNA)的研究进展

非编码小RNA(sncRNA)主要通过抑制性调控靶基因发挥作用。近年的研究发现sncRNA在哺乳动物附睾的各个部位均有表达,并且在调控附睾不同节段的基因表达方面起着非常关键的作用。附睾上皮细胞中sncRNA的多样性及其表达水平的时空差异会影响很多潜在的靶基因,从而调节附睾的生理功能,另外,这些sncRNA还可以通过附睾小体传递给精子,对精子产生一系列的调控作用。综述了附睾中sncRNA表达、sncRNA对附睾和附睾中精子功能调节方面的研究进展,以期为探究sncRNA对雄性生殖细胞调控作用的分子机制提供参考。     

山羊Lcn5的表达特点及其在繁殖器官中定位

为探明附睾视黄酸结合蛋白5(Lcn 5)mRNA在公山羊体组织及不同月龄附睾中的表达特性,及其在睾丸、附睾和精子中的定位。采用实时荧光定量PCR方法检测18种组织和不同月龄附睾Lcn5mRNA表达规律;利用蛋白印迹技术对成年公羊睾丸和附睾Lcn 5蛋白表达进行定量分析;利用免疫组化技术对5月龄公羊睾丸和附睾Lcn 5进行定位,利用免疫荧光技术对精子Lcn 5进行定位。Lcn5mRNA在不同组织中均有表达,其中在性腺中的表达量最高,附睾头附睾尾附睾体睾丸;在5月龄前附睾的Lcn5mRNA表达量随月龄逐渐升高,5月龄时达到最高,随后又逐渐降低。Western blotting结果显示,Lcn 5蛋白表达量:附睾头附睾尾附睾体睾丸,支持了Lcn5mRNA定量的结果。免疫组化结果显示,5月龄山羊在附睾管壁的柱状细胞、纤毛细胞检测到较强Lcn 5蛋白表达的阳性信号,附睾尾管腔检测到较强的阳性信号。在睾丸各级生精细胞中也检测到了微弱阳性信号。Lcn 5蛋白包裹在精子头部顶体帽上。Lcn5在山羊附睾中高度表达,其表达具有时空特异性,推测其在精子成熟和维持正常生理功能方面发挥重要作用,其生物学功能需要进一步深入研究。     

附睾精子抗氧化保护机制研究进展

通过附睾转运,精子经历了一系列的生化和形态变化逐渐成熟,在精子成熟的过程中,其胞质成分含量逐渐减少,因此精子对氧化应激更为敏感,进而导致精子结构和功能受到损害,故而附睾管腔微环境中抗氧化酶的作用非常重要。活性氧(ROS)在精子功能中起着双重作用:生理水平的ROS可促进精子受精前的获能,而过高水平ROS则会导致精子发生氧化损伤。在附睾中清除多余ROS的抗氧化酶主要有超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GPXs)以及过氧化物酶(PRDXs),但是不同年龄的哺乳动物附睾不同部位中各种抗氧化酶的含量均会发生变化,不同抗氧化酶对ROS也具有不同的清除机制。当附睾缺乏某一抗氧化酶时,会使精子DNA受损、精子质量下降,最终导致异常生殖结果增加。因此,哺乳动物附睾中各类抗氧化酶需要相互协调将ROS维持在生理水平,但有关这方面的研究报道较少。附睾上皮细胞分泌的抗氧化酶以附睾小体的形式传递给精子,以清除自身有氧代谢以及异常精子所产生的ROS,进而保护精子正常成熟。作者综述了附睾精子所面临的氧化应激以及附睾中各类抗氧化酶对精子的保护作用。     

SOX5蛋白对公鸡精子发生和精子活力的作用及其定位

本研究旨在探明SOX5蛋白在家禽睾丸发育中的表达规律,分析其在睾丸发育和精子活力调控中的潜在作用。采用RT-qPCR和Western blot技术检测0、5、15、40和60周龄北京油鸡公鸡睾丸中SOX5mRNA和蛋白的表达,并比较其在40周龄正常公鸡与弱精子症公鸡睾丸中的表达差异,通过免疫组化技术对SOX5蛋白在睾丸组织中的表达进行定位。结果表明,SOX5 mRNA和蛋白在不同周龄公鸡的睾丸中均有表达,且差异显著(P0.05)。SOX5mRNA和蛋白表达量趋势一致;随着周龄的增加,其表达呈现先上升后降低的趋势:0和5周龄SOX5蛋白主要在精原细胞和支持细胞中表达;15周龄SOX5蛋白在支持细胞、各级精母细胞、圆形精子细胞和成熟精子中均高表达;40周龄表达量有所降低。SOX5基因和蛋白的表达趋势与公鸡性成熟和繁殖机能衰退的趋势一致,即性成熟前后和繁殖高峰期表达量较高,在性发育前和生精机能减退时表达量较低。此外,40周龄弱精子症公鸡睾丸中,SOX5的表达显著低于正常公鸡(P0.05)。综上表明,SOX5的表达量与公鸡睾丸发育水平和精子活力存在一定的相关,可能对其具有重要调控作用。     

附睾功能及其对精子成熟的影响

哺乳动物的附睾为精子的成熟提供了特殊的微环境，是精子成熟、保护、运输和贮存的部位，在生殖过程中起着重要作用。附睾上皮细胞的合成、分泌、转运等功能使精子发生了一系列结构、生化和功能的改变．从而使精子获得受精和运动能力。作者对附睾的形态结构、功能和对精子成熟影响研究现状等作简要综述。     

催乳素调控禽类繁殖活动的研究现状与展望

本文回顾了近二十年来催乳素在调控家禽繁殖活动方面的研究进展,这些进展包括催乳素分泌的神经内分泌调控机制,催乳素促进就巢发生和抑制产蛋的下丘脑和垂体的中枢内分泌调控机制,以及促进家禽繁殖产蛋的卵泡局部调控机制。催乳素对禽(鸟)类季节性繁殖的调控也分为两种,一种是在依赖年度光照变化调控季节性繁殖的家禽,其年度季节性繁殖周期的形成也是由催乳素和促性腺激素LH分泌的交替消长调控,而对于某些依赖温度而非光照调控的鸟类,催乳素而非LH是启动和终止繁殖季节的主要因素。从催乳素具有抑制和促进禽(鸟)类繁殖产蛋性能的双重调控作用来看,对催乳素作为研究对象而开展的家禽产蛋性能的遗传基础和基因聚合选育研究,还需要继续在理论和实践方面进一步完善。     

附睾特异谷胱甘肽过氧化物酶基因调控的研究进展

附睾特异谷胱甘肽过氧化物酶(epididymal specific glutathione peroxidase,GPx5)可保护精子免受氧化应激,维持精子遗传信息的完整性,促进精子发育成熟,其重要生理功能在提高动物繁殖力上有着不可替代的作用,但其基因调控转录表达的机制尚不明确。现作者主要对基因表达特性、表达调控和转录调控等内容进行综述。     

家禽睾丸主要生殖激素受体的研究进展

睾丸的主要功能是制造精子,而睾丸中生殖激素与其受体的结合又影响着精子的发生和成熟,因此家禽睾丸中生殖激素受体的研究对提高其受精率具有十分重要的意义。本文主要阐述四类生殖激素受体——促卵泡激素受体、促黄体激素受体、雄激素受体和雌激素受体的研究进展,为提高家禽繁殖力,推进家禽养殖业发展提供一定的参考。     

L—PGDS基因在大鼠睾丸与附睾中的表达

选用2月龄SD大鼠睾丸与附睾总RNA为模板，运用RT—PCR和蛋白质印迹技术探讨了Lipocalin型前列腺素D合成酶(L-PGDS)基因在大鼠睾丸与附睾头、附睾体和附睾尾中的表达及其差异。结果表明，L—PGDS基因在大鼠睾丸中没有表达，在附睾中有较强表达，附睾头表达量最高，附睾体次之，附睾尾最低，表达量存在差异。提示，L-PGDS在精子成熟过程中起着一定的作用。     

雄激素受体基因在绵羊公羔不同组织中的表达特性及性腺中的定位

旨在研究雄激素受体(Androgen receptors,ARs)基因在绵羊公羔不同组织中的表达特性及在性腺中的定位。本研究以QRT-PCR法检测各组织ARmRNA表达规律,免疫组化技术对睾丸和附睾中AR进行定位分析,蛋白质印迹技术对AR蛋白表达量进行定性研究。结果显示:(1)AR在不同组织中均有不同程度的表达,其中在睾丸和心脏中大量表达;(2)在附睾头、体、尾部单层柱状上皮细胞和间质细胞中检测到较弱的阳性信号;睾丸组织间质细胞、精原细胞、管周肌样细胞检测到强阳性信号。综上表明:AR在绵羊不同组织中广泛表达,AR在性腺的发育和精子的成熟过程以及对心脏的保护方面发挥重要作用,其作用机制需进一步研究。     

哺乳动物附睾特异GPx5功能及其调节机制的研究进展

精子在附睾中的成熟过程是哺乳动物雄配子获得受精能力的关键。谷胱甘肽过氧化物酶-5（glutathione peroxidase-5,GPx5）作为附睾特异性表达的抗氧化酶具有强抗氧化作用,可调节附睾微环境中的活性氧浓度,保护精子免受脂质过氧化损伤,以维持精子DNA完整性。GPx5还可能对精子活力和顶体反应产生一定影响。GPx5基因的染色体定位及其外显子数存在一定的种间差异,其在不同物种、部位和发育期的表达具有特异性,受雄激素、PEA3因子和ETV4家族等调节。作者就哺乳动物附睾特异GPx5基因的结构与定位、表达特性、功能及其调节机制的研究进展进行综述,以期为进一步研究精子抗氧化机制和提高雄性动物繁殖力提供理论依据。     

Comparative rna-seq analysis of region-specific miRNA expression in the epididymis of cattleyak

The epididymis is the site of post-testicular sperm maturation, which constitutes the acquisition of sperm motility and the ability to recognize and fertilize oocytes. The role of miRNA in male reproductive system, including the control of different steps leading to proper fertilization such as gametogenesis, sperm maturation and maintenance of male fertility where the deletion of Dicer in mouse germ cells led to infertility, has been demonstrated. The identification of miRNA expression in a region-specific manner will therefore provide valuable insight into the functional differences between the regions of the epididymis. In this study, we employed RNA-seq technology to explore the expression pattern of miRNAs and establish some miRNAs of significant interest with regard to epididymal sperm maturation in the CY epididymis. We identified a total of 431 DE known miRNAs; 119, 185 and 127 DE miRNAs were detected for caput versus corpus, corpus versus cauda and caput versus cauda region pairs, respectively. Our results demonstrate region-specific miRNA expression in the CY epididymis. The GO and KEGG enrichment for the predicted target genes indicated the functional values of miRNAs. Furthermore, we observed that the expression of miR-200a was downregulated in the caput, compared with cauda. Since the family of miR-200 has previously been suggested to contribute to the distinct physiological function of sperm maturation in epididymis of adult rat, we speculate that the downregulation of miR-200a in CY caput epididymis may play an important role of sperm maturation in the epididymis of CY. Therefore, our findings may not only increase our understanding of the molecular mechanisms regulated by the miRNA functions in region-specific miRNA expression in the CY epididymis, it could provide a valuable information to understand the mechanism of male infertility of CY.     

GPx6在大白猪附睾细胞中的表达及其与繁殖性能的相关性

本试验旨在研究谷胱甘肽过氧化物酶6（glutathione peroxidase 6,GPx6）在大白公猪附睾细胞中的表达和定位,并探究其与公猪繁殖性能的相关性。选取15月龄的公猪和母猪各3头,取睾丸、附睾、前列腺、尿道球腺、精囊腺、卵巢和输卵管,提取蛋白,通过蛋白免疫印迹和免疫组化（IHC）检测组织和细胞中GPx6蛋白的表达和定位;根据评定大白公猪受精能力的数学模型,按照公猪配种胎次≥20胎、3次配种公猪为同一头的标准,筛选并采集符合要求的20头公猪精液,同时,统计相对应的1 279头母猪的生产成绩,计算得到公猪繁殖性能指标（包括窝产总仔数、窝产活仔数、分娩率和繁殖力）。提取精子蛋白和精清蛋白,通过BCA和ELISA检测精子和精清中GPx6蛋白的含量。使用SPSS软件的独立样本t检验及单因素方差分析（one-way ANOVA）,进行差异显著性分析,用双变量Pearson进行相关性分析,P<0.05表示差异或相关显著。结果表明,GPx6蛋白在附睾中高表达,IHC结果显示,GPx6蛋白在附睾的顶细胞、基底细胞、晕细胞、主细胞和精子中表达,在肌样细胞中不表达;精清蛋白中GPx6的含量是精子蛋白的7倍,精液中GPx6蛋白的含量与窝产活仔数、窝产总仔数呈负相关关系。结果提示,GPx6在附睾的顶细胞、基底细胞、晕细胞、主细胞和精子中表达,且其在精液中的含量会影响公猪的繁殖性能,这为GPx6对公猪受精能力影响的研究奠定了理论和试验基础。     

不同营养成分对公猪精液品质的影响

公猪优良精液的产生与必要的营养供给是密不可分的。为探讨鸡蛋、豆粕、胡萝卜三种营养物质对公猪精液品质的影响,本试验通过评价精液采集量、精子密度、活力及畸形率等指标,来判定其对猪精液的影响。试验表明,公猪日粮中饲喂豆粕和鸡蛋对公猪精液的采集量,活力均有显著的提高;饲喂胡萝卜对公猪精液的采集量,活力无明显差异。为公猪饲养过程中提供一些实际的生产指导。     

热应激对雄性哺乳动物精液品质的影响机制及热应激公猪的营养调控

当环境温度超过机体热调控上限时,动物体将发生热应激.热应激对雄性哺乳动物的性欲、精液品质等均有不利影响,可使精子密度和精子活力下降、精子畸形率上升等.本文总结了热应激对雄性哺乳动物精液品质影响的作用机制以及热应激公猪的营养调控,以期为相关研究和生产实践提供参考.     

绵羊精子细胞核质转运蛋白KPNA4的研究

旨在对绵羊附睾头、体和尾部的精子进行蛋白质组学分析,获得差异表达蛋白,对数据进行功能富集分析,挖掘精子发生/成熟关键蛋白质。本研究选择12月龄左右健康的3只雄性湖羊为试验动物,分离附睾并按区域收集精子,3组样本(附睾头部组、附睾体部组和附睾尾部组),每组3个生物学重复,共计9例绵羊精子细胞样本。基于TMT标定定量蛋白质组学分析和R语言等工具,在获取的差异表达蛋白中进行GO和KEGG富集分析,并利用蛋白质免疫印迹(Western bolt)、免疫荧光(immunofluorescence)和流式细胞术(flow cytometry)试验验证结果的可靠性。从22 841个唯一性肽中鉴定到差异蛋白质616种,其中,尾vs头组鉴定出309个差异表达蛋白(上调213个,下调96个);尾vs.体组鉴定出167个差异表达蛋白(上调107个,下调60个);体vs头组鉴定出140个差异表达蛋白(上调88个,下调52个)。根据差异倍数与蛋白质功能,筛选出可能与精子成熟、核质物质转运相关的关键蛋白-KPNA4。本研究揭示了绵羊附睾不同部位精子的特点与差异,这些数据为研究雄性绵羊的生殖机制和精子成熟提供了丰富的资源。     

Semen Collection, Examination and Spermiogram in Ostriches

The level of fertility in the male ostrich exerts considerable influence on the efficiency of the fertilization procedure, and thus also on reproductive performance. The determination of the reproductive capacity is of particular interest with regard to the selection of single individuals for optimizing reproduction ratios. Although the breeding and raising of ostriches has become increasingly important in many countries, little research has been completed on reproductive parameters and factors that may possibly influence them. This study presents observations made concerning the quantity and quality of sperm as found in the spermatological testing of 411 ejaculate samples taken from male ostriches on two farms in Namibia. The semen volume varied between 0.1 and 1.5 ml (mean, 0.64 ml). Normal ejaculate colours ranged from white to ivory; the consistency ranged from thin creamy to viscous. The measured pH values lay between 6.4 and 8.0 (mean, 7.3). Microscopic investigations revealed sperm concentrations of 8.9–78.1 million/μl and individual sperm motility from 42 to 96% (mean, 78%). No mass motility was detectable in 42% of the ejaculates; weak mass motility was found in 46%, and clear mass movements were to be found in only 12% of samples. Regarding the morphology of the sperm, 5 to 26% were abnormal (mean, 17%) and 4 to 28% (mean, 20%) were dead. Seasonal patterns of sperm concentration and the influence of frequency of semen collection were investigated in a group of 56 healthy male ostriches. Peak sperm concentrations were found at the beginning of the breeding season in spring; the lowest values were found at the end of the breeding season in autumn. The highest quality ejaculate was obtained from those males whose semen was collected once a week. The results of this study provide fundamental data for the establishment of minimum quality requirements for ostrich sperm to be met by individuals receiving certification as breeding animals and for the selection of suitable males for use in artificial insemination.     

Research Progress on Genetic Breeding and Reproduction of Domestic and International Rabbits in 2019

The rabbit is a kind of grain-saving and small herbivore animal.Rabbit breeding which is low investment and obvious efficiency has become an advantageous industry of increasing income of rural farmers and poverty alleviation.In recent years,with the development of life science,great progress has been made in rabbit genetic breeding at home and abroad.In this paper,the research progress in genetic breeding and reproduction of rabbits in 2019 are reviewed from the traditional breeding,molecular breeding and reproduction of rabbits.For abroad,lots of researches involved in traditional breeding,molecular breeding and reproduction were done.Effects of selection,environment and additives on productive performance had been analyzed in traditional breeding.Molecular breeding mainly focused on reproductive performance among which there were more studies on litter size and spermatine.Secondly,the related genes of growth,meat quality and hair color were studied.The research on reproduction mainly focuses on the preservation method of male rabbit semen and the effects of additives on rabbit semen quality,litter size and conception rate.In China,genetic breeding and reproduction research on rabbit also mainly included traditional breeding,molecular breeding and reproduction,but the focus of domestic research was molecular breeding,among which breed and genetic diversity and functional genes of fur trait were mainly researched using high-throughput sequencing,gene cloning and gene editing to screen the important functional genes and regulatory networks related to research traits.However,compared with molecular breeding,there are fewer studies on the traits evaluation of traditional breeding and rabbit reproduction.This review can provide references for rabbit breeding and production.