棕熊精子体外获能及异种穿卵的超微结构研究

对棕熊精子体外获能前后和异种穿卵的超微结构观察表明，棕熊精子全长７７μｍ，由头、颈和尾３部分组成，头部长７．３ｍ，宽２．５μｍ，主要由核、顶体及顶体后区组成；颈部由中心粒和９条纵行分节的节柱组成；尾部全长６８．２μｍ，其中中段长１３．２１ｍ，线粒体为６５－６８旋，主段中央为“９＋２”的微管结构，基外方被９条致密纤维和纤维鞘包裹。精子获能前群集成簇，运动缓慢；获能后精子呈超激活运动。获能的精子质膜膨     

贵州小型猪精子显微结构研究

应用光学显微镜、透射电镜和扫描电镜对贵州小型猪精子的形态和结构进行了研究.结果表明,贵州小型猪精子由头、尾二部分组成.精子全长57.00±1.69 μm;头部呈椭圆形体,长为9.49±0.40 μm;尾部由颈、中、主和末段组成;尾部中央为9×2 2排列模式的轴丝,轴丝外围依次是外周致密纤维、线粒体螺旋鞘、纤维鞘、细胞膜;线粒体螺旋鞘仅存在于中段,外周致密纤维从主段到末段逐渐变细,并在主段近末端处分批终止,末段外围仅保留纤维鞘、细胞膜.贵州小型猪精子组织结构与人类接近,是研究人类男性生殖系统疾病较理想的动物模型.     

棕熊精子体外获能及异种穿卵的超微结构研究

对棕熊精子体外获能前后和异种穿卵的超微结构观察表明,棕熊精子全长77μm,由头、颈和尾3部分组成.头部长7.3μm,宽2.5μm,主要由核、顶体及顶体后区组成;颈部由中心粒和9条纵行分节的节往组成;尾部全长68.2μm,其中中段长13.2μm,线粒体为65～68旋,主段中央为“9 2”的微管结构,其外方被9条致密纤维和纤维鞘包裹.精子获能前群集成簇,运动缓慢;获能后精子呈超激活运动.获能的精子质膜膨胀,顶体外膜囊泡化,引起顶体反应,质膜并未参加囊泡化.顶体反应完成后,仅有顶体内膜包在精子核膜的外面.棕熊精子与仓鼠的卵相互作用,多以赤道段和顶体后区附着于卵膜.     

鹅精子电子显微镜超微结构初步探讨

鹅精子头部为棒状.电子扫描镜观察头和尾之间无明显界限。电子透射观察发现头部核质(nu-clear substance)中有空泡(vacuole),由近中心粒所形成的九组三连筒状结构与尾部纤丝系统所成的夹角约为20°,尾部中段有一个约0.83微米的无中心纤丝区,精子头部长10.40±2.01微米,精子全长75.10±2.45微米。     

德国牧羊犬精子超微结构及超低温冷冻对精子超微结构的影响

为了研究超低温冷冻前后德国牧羊犬精子超微结构的变化,采用扫描电镜和透射电镜对冷冻前后德国牧羊犬精子超微结构变化进行研究,结果显示在扫描电镜下,鲜精整体呈“蝌蚪状”,头部呈扁卵圆形.在透射电镜下精子纵切面头部呈“楔形”,头部细胞膜由外向内分别由质膜、顶体外膜、项体内膜和核膜组成.精子尾部中段横切面由外向内可见线粒体鞘膜、9束外周致密纤维,9对轴丝和2根中央微管.精子头长约为5.40 μm,头宽3.26 μm,颈长1.25 μm,颈宽0.55 μm,尾部中段长11.34 μm,中段直径0.87 μm,尾部长55.70 μm,线粒体螺旋数为44旋.鲜精和冷冻前精子头部、颈部和尾部形态变化差异均不显著(P＞0.05).超低温冷冻后精子头部、颈部和尾部发生形态变化的精子数极显著高于鲜精和冻前精子数(P＜0.01).冻后顶体发生明显形态变化的精子占54.21％,高于发生颈部形态变化(10.61％)和尾部形态变化(28.83％)的精子数.结果表明鲜精和冻前精子形态变化不显著,而超低温冷冻后精子头部、颈部和尾部发生形态变化的精子数显著高于鲜精和冻前精子数.     

冷冻保存绵羊精子质膜完整性检测的研究

为了探讨冷冻保存对绵羊精子质膜完整性的影响,研究采用低渗肿胀—伊红拒染(HOS-EY)法检测冷冻保存前后绵羊精子质膜的完整性。实验设计了5个HOS-EY低渗染液浓度梯度组合(110、130、1501、70、190mOsm),按精子尾部肿胀与否和头部着色与否区分为4种类型进行统计。结果表明:绵羊冻融精子头膜尾膜均完整的Ⅳ型精子率(22.36%)和头膜尾膜均损伤的Ⅰ型精子率(47.4%)与鲜精比较分别极显著减少(58.64%)和增多(20.94%)(P<0.01),表明冷冻-解冻过程对精子膜造成严重损伤;头膜损伤-尾膜完整的Ⅱ型精子率(28.82%)在冷冻过程中极显著增多(16.72%)(P<0.01),头膜完整-尾膜损伤的Ⅲ型精子在冷冻组发生率(1.72%)显著减少(2.58%)(P<0.05),表明精子头膜或尾膜的损伤是独立发生的,且头膜损伤易于尾膜,5种HOS-EY低渗染液中150 mOsm组对精子尾部肿胀的检测效果最好。     

鸵鸟精子的超微结构

为了丰富鸵鸟精子的形态资料和为提高鸵鸟的人工授精技术提供参考,采用透射电镜和扫描电镜对鸵鸟精子的超微结构进行观察,并且与其它非雀形目鸟类以及哺乳动物的精子结构进行了比较。结果表明,鸵鸟精子呈细长的圆柱状,分为头部、颈段、中段、尾段、末段,其结构与美洲鸵和Tinamou精子相似。鸵鸟精子头部由顶体和精子核组成,精子核中有一条顶体刺。鸵鸟精子没有明显的颈部。中段的外周环绕着线粒体鞘。主段的中央为轴丝,轴丝外周有纤维鞘。鸵鸟精子的超微结构与美洲鸵和tinamou相似,但是与其它非雀形目的鸟类和哺乳动物的精子有很大的区别。     

低温保存对绵羊精子质膜完整性的影响

为了探讨低温保存对绵羊精子质膜完整性的影响,研究采用低渗肿胀-伊红拒染(HOS-EY)法检测低温保存前后绵羊精子头膜和尾膜的完整性。试验设计了5个HOS-EY低渗染液浓度梯度组合(110,130,150,170,190 mOsm),按精子尾部肿胀与否和头部着色与否分为4种类型进行统计。结果表明:低温保存后,绵羊精子头膜、尾膜均完整的Ⅳ型精子发生率和头膜、尾膜均损伤的Ⅰ型精子发生率与鲜精相比分别极显著减少和增多(P<0.01),说明低温保存过程对精子质膜完整性有极显著影响;在低温保存过程中头膜损伤-尾膜完整的Ⅱ型精子发生率极显著增多(P<0.01),而头膜完整-尾膜损伤的Ⅲ型精子发生率显著减少(P<0.05),说明精子头膜或尾膜的损伤是独立发生的,且头膜比尾膜更易损伤。5种低渗液中,150 mOsm染液检测效果最好。     

高原地区成年藏绵羊和小尾寒羊附睾组织中eNOS差异表达研究

采用免疫组化SP法结合IPP统计分析法,比较高原地区成年藏绵羊和小尾寒羊附睾组织中eNOS的定位及分布特征。平均光密度数据统计表明,eNOS的平均光密度值在藏绵羊附睾尾部显著高于附睾头和附睾体(P0.05),藏绵羊附睾头和附睾体之间无显著差异(P0.05),但在小尾寒羊附睾体显著高于附睾头(P0.05)。小尾寒羊附睾体的平均光密度值显著高于藏绵羊(P0.05),附睾头和附睾尾之间无显著差异(P0.05)。结果表明,eNOS的活性在附睾尾部最高,提示NO与精子运输相关,对于高原地区不同品种绵羊附睾中eNOS分布较大差异的原因尚需深入研究。     

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。     

Ultrastructure of the spermatozoa of the domestic duck (Anas platyrhynchos sp.)

The ultrastructure of the spermatozoa of the domestic duck (Anas platyrhynchos sp.) was analysed by transmission and scanning electron microscopies and compared with the results obtained in preliminary studies involving other non-passerine birds. The spermatozoa were characterised by the presence of a short head, short midpiece and long principal piece. The head consisted of a reduced acrosome that contained moderately electron-dense homogenous material. The implantation fossa was observed between the base of the nucleus and the proximal centriole. The midpiece contained electron-dense material associated with the proximal centriole and nuclear membrane, and a long distal centriole surrounded throughout its length by 11-12 elliptical mitochondria. A dense annulus separating the midpiece from the principal piece was visible. Posterior to the annulus, the axoneme was formed surrounded by a dense fibrous sheath, representing the principal piece or flagellum, which was a long segment with a smooth surface and a smaller diameter than the midpiece. The spermatozoon of the domestic duck resembles that of other non-passerine birds, corresponding to a basic type of spermatozoon similar to that of reptiles, called sauropsid type.     

The Structure of the Spermatozoon of Liza Dumerili (Teleostei) With Notes on Spermiogenesis

The structure of the sperm of Liza dumerili was studied by light microscopy and scanning and transmission electron microscopy.

The sperm have a kidney-shaped nucleus. The “hilus” faces posteriorly and the proximal centriole is situated within the nuclear pit. Nuclear rotation takes place during spermiogenesis and the sperm are therefore related to those of other teleosts showing Type I spermiogenesis. No acrosome is evident. The midpiece is small but distinct and contains four spherical mitochondria. The flagellum enters the midpiece eccentrically and the axoneme runs obliquely towards the distal centriole to which it is connected. The flagellum conforms to the 9 + 2 flagellar pattern.     

Differences Between High‐ and Low‐Motility Buffalo Sperm Identified by Comparative Proteomics

This study was undertaken to investigate differences in protein expression between high‐ and low‐motility sperm of swamp buffalo. The research used two‐dimensional gel electrophoresis (2DE) coupled to matrix‐assisted laser desorption/ionization time‐of‐flight tandem mass spectrometry (MALDI‐TOF/TOF‐MS) to analyse the different proteins. The results showed 18 different expression protein spots between high‐ and low‐motility buffalo sperm; eight of these proteins were up‐regulated in low‐motility sperm, five were down‐regulated, one deleted and four proteins specifically expressed. Finally, four proteins were successfully identified by MS as belonging to three unique proteins; they are outer dense fibre of sperm tails protein 2 (ODF2), ATP synthase subunit alpha (ATP5A1) and succinyl‐CoA synthetase subunit beta (SUCLG2). In summary, these results help to develop an understanding of the molecular mechanisms associated with low‐motility sperm and provide clues for finding molecular markers associated with sperm motility.     

Comparison of the structure of chinchilla sperm isolated from semen and from the tail of the epididymis

Sperm cells isolated from the tail of the epididymis and from the semen of the same individuals were analysed. The use of silver nitrate to stain sperm cells isolated from the tail of the epididymis made it possible to identify structures that were not visible in the sperm from semen. Silver nitrate very clearly distinguished the acrosomal and distal parts of the sperm head. Following silver nitrate staining, the sperm isolated from the tail of the epididymis were characterized by dark ‘collars’ in the distal part of the head. These ‘collars’ are not visible in the sperm cells isolated from semen. The results of the study indicate differences in the dimensions of sperm isolated from the tail of the epididymis and sperm in semen. Sperm isolated from the tail of the epididymis had smaller heads, despite their longer length, and had longer midpieces and tails than ejaculate sperm. Silver nitrate staining is a simple and fast technique. Silver nitrate makes it possible to identify the acrosome and post-acrosomal region of the sperm head and to clearly identify the midpiece. Therefore, it can be successfully used to supplement routine techniques for evaluating sperm morphology or as an independent technique.     

马鹿冻精解冻后精子超微结构观察

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。     

马鹿精子形态与超微结构研究

文中对马鹿(Cervus elaphus)精子形态与超微结构进行了研究。马鹿精子经固定、脱水、置换、包埋和聚合,用超薄切片机切片,再用醋酸双氧铀、柠檬酸铅染片,最后于透射电镜下观察其超微结构的变化。观察结果表明:马鹿精子全长(58.75±2.35)μm,由头部(8.93±0.24)μm、颈部(1.00±0.16)μm和尾部(48.18±1.18)μm三部分组成;头部呈扁卵圆状,绝大部分被浓缩且电子密度较高的精核所占据,顶体似帽状扣在精核之上,约占头部的2/3,其前部较为膨大;颈部位于头部和尾部之间,这个区域较短;尾部可分为中段、主段和终段。马鹿尾部轴丝的结构类型为"9+2";微管结构类型为"9+9+2"。     

影响意大利水牛精液品质的相关因素分析

本研究旨在通过对意大利水牛精液品质分析、睾丸周径测量、精浆中的氧化应激水平检测和精子活力相关基因表达情况来探究影响精液质量的相关因素。试验检测了6头意大利水牛精液的活力、畸形率、采精量,并在测量其阴囊周径后进行相关性分析;检测了意大利水牛精浆中氧化应激水平指标（丙二醛（MDA）、总超氧化物歧化酶（T-SOD）、谷胱甘肽过氧化物酶（GSH-Px））并进行了相关性分析;分析了意大利水牛精子β-微管蛋白-2c（TUBB2C）、外周致密纤维2（ODF2）、筑丝蛋白2（TEKT2）、筑丝蛋白4（TEKT4）基因的表达定量及相关性。结果显示,意大利水牛阴囊周径与精液产量、活力、畸形率之间相关系数分别为0.423（P>0.05）、0.750（P<0.01）、-0.827（P<0.01）,即阴囊周径与精子活力呈显著正相关关系、与畸形率呈显著负相关关系;意大利水牛精子活力与MDA、T-SOD、GSH-Px指标之间相关系数分别为-0.522（P<0.05）、0.333（P>0.05）、0.474（P<0.05）,即精子活力与MDA含量之间存在显著负相关关系,与GSH-Px活性之间存在显著正相关关系,与T-SOD活性相关性不显著;意大利水牛精子活力与TUBB2C、ODF2、TEKT2、TEKT4基因指标之间相关系数分别为0.930（P<0.01）、0.726（P<0.01）、0.924（P<0.01）、0.839（P<0.01）,即精子活力与以上基因表达量存在显著正相关关系。本研究结果为了解影响意大利水牛精液质量的因素提供参考,也为意大利种公水牛的筛选提供理论依据。     

Analysis of Related Factors in Semen Quality of Italian Buffaloes

This study was aimed to investigate the factors affecting semen quality by analyzing the semen quality of Italian buffaloes,measuring the circumference of the testis,the level of oxidative stress in seminal plasma and the expression of genes related to sperm motility.In this study,the sperm vitality,deformity rate and semen collection of 6 Italian buffaloes were tested,and the correlation analysis was carried out after measuring scrotal circumference.The oxidative stress level of seminal plasma in Italian buffaloes was tested,including malondialdehyde (MDA),total superoxide dismutase (T-SOD) and glutathione peroxidation (GSH-Px).The expression of β-tublin-2c (TUBB2C),outer dense fibres 2 (ODF2),tektin-2 (TEKT2),tektin-4 (TEKT4) genes and their correlation with sperm in Italian buffaloes were analyzed.The results showed that the correlation between scrotal circumference and semen yield,vigor and deformity rate was found,the coefficients were 0.423 (P>0.05),0.750 (P<0.01) and -0.827 (P<0.01),which showed a significant positive correlation between scrotal circumference and sperm motility,and a significant negative correlation with the deformity rate.The oxidative stress level in seminal plasma of Italian buffaloes was tested,the correlation coefficients of MDA,T-SOD and GSH-Px with sperm motility were -0.522 (P<0.05),0.333 (P>0.05) and 0.474 (P<0.05),respectively.There was a significant negative correlation between sperm motility and MDA content,a significant positive correlation between GSH-Px activity and sperm motility,no significant correlation with T-SOD activity.The correlation coefficients between sperm motility and TUBB2C,ODF2,TEKT2 and TEKT4 genes were 0.930 (P<0.01),0.726 (P<0.01),0.924(P<0.01) and 0.839 (P<0.01),respectively.There was a significant positive correlation between sperm motility and the expression of four genes.The results provided a reference for understanding the factors affecting the semen quality of Italian buffaloes,and also provided a theoretical basis for the selection of Italian bull.     

The relationship between sperm morphology and in vitro fertilization ability in mice

In vitro fertilization (IVF) is widely used in reproduction research, but the sperm of some inbred strains of mice yield low fertilization rates in IVF. To determine the cause of this problem, we examined the effect of epididymal sperm morphology, in particular, tail bending and the presence and type of cytoplasmic droplet (CD), on fertilizability in vitro. Sperm suspensions were obtained from the following five strains: C57BL/6J, BALB/cA, C3H/HeN, DBA/2J, and 129 x 1/SvJ. The sperm were fixed in 10% formalin and three parts of the sperm, namely the head, tail, and CD, were examined. We recorded the proportion of abnormal sperm heads and hairpins at the neck; tails were categorized as straight, proximal bent, or distal bent; and the CDs were categorized as none, light-type, and heavy-type. Based on these parameters, we determined the correlations between sperm morphology and fertilizability in vitro, as judged by IVF using ICR oocytes. The proportion of sperm with a hairpin neck was higher in strain C57BL/6J, while abnormal head morphology occurred significantly more often in strain BALB/cA. The percentage of sperm with a proximal bent tail was highest in strain DBA/2J and lowest in strain 129 x 1/SvJ. A heavy-type CD was observed more frequently in the 129 x 1/SvJ and C57BL/6J strains than in the other three strains in which a light-type CD predominated. The rank order of the fertilization rates was 129 x 1/SvJ < C57BL/6J < C3H/HeN < BALB/cA < DBA/2J. In addition, fertilization rate was positively correlated with a proximal bent tail, but negatively correlated with a heavy-type CD and distal bent tail. This new classification system establishes that the morphological characteristics of epididymal sperm differ among inbred strains of mice and that tail and CD morphology are closely related to fertilization ability in IVF. Thus, our results provide a novel method for assessing the quality of mouse sperm used for IVF.     

The Hereditary 'Short Tail' Sperm Defect - A New Reproductive Problem in Yorkshire Boars

This study describes a new sperm defect in Yorkshire boars. The length of the sperm tail is markedly reduced, resulting in a total immotility in all spermatozoa. At transmission electron microscopy level, the morphology of the midpiece microtubular components area is seriously affected. This boar sperm defect differs from the ‘tail stump’ defect observed in bulls, the tails being longer in most spermatozoa than those found in affected bulls. Therefore, the term ‘short tail’ sperm defect is more adequate. The authors observed the first case in 1987. In 1998, this defect became a noteworthy reproductive problem, when it was observed in nine boars intended for breeding. In one litter, three littermates were affected with the ‘short tail’ sperm defect. At the present time the authors believe that the defect is recessively inherited.