莱氏拟乌贼纳精囊超显微结构

本研究采用组织切片和透射电镜技术，对莱氏拟乌贼(Sepioteuthis lessoniana)纳精囊进行超显微结构分析，并通过扫描电镜和投射电镜技术观察纳精囊中精子的形态构造。结果显示，莱氏拟乌贼口膜具有1个椭圆形的纳精囊，呈乳白色，交配后雌体的口膜内侧散布大量精子囊。纳精囊由囊壁组织、中心管腔和储精小囊组成，储精小囊数量较多，少数呈圆形或椭圆形，2个或多个储精小囊形成中心管腔，二者的囊腔中均分布大量精子。储精小囊和中心管腔由上皮细胞组成，细胞间排列紧密，其细胞核较大，细胞质中含有多种细胞器，包括线粒体、内质网、高尔基复合体和囊泡等；中心管腔中精子大多排列紧密有序，精子头部朝向纤毛，鞭毛位于囊腔中间。莱氏拟乌贼的精子头部细长，由圆屋顶形的顶体和细长圆柱形的精核组成，线粒体距位于精核后端，尾部为细长鞭毛。研究表明，莱氏拟乌贼纳精囊具有储存精子的功能。其中，储精小囊的上皮细胞具备一定分泌功能，其分泌物质能吸引精子进入纳精囊。     

金乌贼纳精囊组织结构及其精子的储存和利用

为揭示金乌贼精子进入纳精囊及产卵过程中的精子利用方式,丰富金乌贼繁殖生物学研究内容,本研究利用实验生态学和组织切片技术,检测了交配后不同时间段雌性口膜表面精子囊和纳精囊中精子数量的变化,观察分析了雌性金乌贼纳精囊的组织结构。结果显示,金乌贼纳精囊位于繁殖期雌性个体口膜腹面的突起处,共1对。纳精囊开口于口膜内表面,通过一根中央管连通整个纳精囊。中央管内壁含有大量褶皱和纤毛。在中央管两端,有12~20个储精小囊与之相连。储精小囊四周具有发达的环肌,其中储存有大量精子,并且大部分精子头部均朝向腔室内壁。完成一次交配后,雌性金乌贼对精子囊和纳精囊中精子的利用可以分为三个阶段,主要利用精子囊中的精子(交配后1~2 d);由利用精子囊中的精子向纳精囊中的精子过渡(交配后2~3 d);主要利用纳精囊中的精子(交配后3 d以上)。研究表明,从精子囊释放出的精子进入雌性口膜表面的褶皱中,通过自身运动到达纳精囊。进入纳精囊的精子通过自身运动及中央管内壁纤毛的摆动进入储精小囊,其中大部分精子头部朝向储精小囊内壁有规律地分布。在产卵过程中,雌性优先利用精子囊中的精子,而在精子囊中精子不足时,纳精囊通过肌肉收缩以及纤毛摆动将其中的精子逐渐释放出来,卵子在雌性口膜附近完成体外受精。     

嘉庚蛸生殖系统结构观察

采用解剖和组织切片技术对嘉庚蛸生殖系统结构进行了系统研究。雄性生殖系统除已报道的精巢、输精管、精囊、摄护腺、精荚囊和阴茎等部分之外,新发现了盲囊结构。精巢属于小叶型精巢,由精巢壁和实质部分组成。精囊分为前、中、后3段,前段与输精管相连,后段与摄护腺相连。精囊壁组成复杂,有两个舌状突起伸入囊腔中,两个舌状突起结构从精囊中段渐行至精囊后段,一个逐渐膨大,另外一个逐渐缩小,精囊壁及舌状突起上密布腺体细胞,分泌作用旺盛。精荚及胶质尾巴均在摄护腺中形成,在精荚囊中浓缩、储存。雌性生殖系统包括卵巢、输卵管和输卵管腺等部分。输卵管较长,开口于漏斗口处。输卵管腺着生在输卵管上,结构复杂,内部有纳精囊结构。     

环境条件对中国对虾交尾影响的试验观察↑（*）

本文报告了在室内条件下中国对虾雄性生殖系统不同部位精子形态、激活反应及外界环境对对虾交尾影响等的试验观察。雄性生殖系统被分为３部分：精巢（Ｔ）、输精管和精荚（ＴＡ）;输精管又分为前、中、后３段,其中的前（ＡＶＤ）、中（ＭＶＤ）、后（ＰＶＤ）３部分以及精巢和精荚被分别匀浆取样镜检,观察精子形态。结果表明：中国对虾雄性精巢、输精管、精荚及雌虾纳精囊内精子的形态、对卵水的激活反应能力有所差异,是一逐渐成熟过程;雄虾具有多次交尾能力,不同环境条件对中国对虾的交尾影响明显;当雌虾比例较高时交尾率也高;不同对虾密度对交尾影响不明显     

中国对虾卵水的特性和精子的应答

研究了中国对虾卵水的采集、保存方法、紫外吸收特性和有效期等方面的内容，并应用卵水研究了交尾期虾、雌虾及产卵期雌虾精荚或纳精囊中精子对卵水的应答开始时间、必要的反应时间等响应卵水的时间特性以及精子的存活期等。结果表明，卵水经液氮保存或先经液氮后转入普通冰柜保持冻结状态７个月后诱导精子激活效力无显著差异。精子在自然温度（１０℃）普通海水中可在１０ｈ内保持对卵水的响应能力。交尾期精子最初响应卵水有５￣１     

澳洲淡水红螯虾繁殖特性及精荚显微结构的研究

报道了澳洲淡水红螯虾的繁殖特性及精荚显微结构，澳洲淡水红螯虾繁殖能力强，最适繁殖温度为２８￣３０℃，雄虾在第五对卡足左部内侧有雄性生殖孔，呈小球状。由雄虾射出的精荚中含有半透明管状结构，内含大量成熟精子，精子有一个椎形的头基部和细长的棘状部，雌雄交配过程是雄虾将成熟精荚射到雌虾蝮面，雌虾在２４ｈ内排卵并行受精。     

头足类精子超微结构研究进展

精子作为物种生殖生物学、受精生物学研究的重要内容之一,其发生及超微结构具有属种特异性,是系统分类、亲缘关系和生殖进化的重要依据。头足类是重要的海洋生物,在海洋生态系统中占有重要的地位,其精子发生主要经历顶体的演变、精核的建成、核后端线粒体的系统演变等过程,不同属种的成熟精子超微结构存在显著差异。八腕目属种精子顶体呈螺旋状钻头形,精核为直核,线粒体于尾部形成线粒体鞘;乌贼目属种精子顶体呈圆屋顶形或圆锥形,精核稍弯,线粒体裹于线粒体距中;幽灵蛸目的幽灵蛸和旋壳乌贼目的旋壳乌贼精核均为直核,线粒体围绕鞭毛形成线粒体袖套,前者顶体球形,后者顶体圆锥形。其中,线粒体距、线粒体袖套、线粒体鞘等头足类精子的线粒体系统变化,是精子对受精环境的适应,也是头足类不同属种生殖进化的演化产物。     

大菱鲆成熟精子、卵子及精子入卵早期过程的电镜观察

通过人工授精和电镜技术,观察和描述了大菱鲆Scophthalmus maximus成熟精卵的形态、精子入卵的早期过程及精子入卵过程发生的一系列形态结构变化。大菱鲆卵子表面均匀地布满纵横交错的网纹,并整齐地分布许多微小孔,在动物极有一个受精孔。大菱鲆精子为无顶体类精子,由头部、中片和尾部三部分组成,中片有9～14个圆形线粒体。大菱鲆精子入卵速度非常快,授精后0～5s已经有精子通过受精孔(Micropyle)进入卵子。精子入卵过程还伴随着其他一些结构的变化,如精孔管管壁边缘由锯齿状变为平滑的环状等。     

[4]东海三疣梭子蟹精子发生及精荚形成

为探究东海三疣梭子蟹(Portunus trituberculatus Miers,1876)雄性群体的性腺发育规律,通过定期采样,运用形态学、组织学方法对东海三疣梭子蟹精子发生及精荚形成过程进行了研究,并结合精小叶生殖节段法对处于不同发育阶段的精巢进行了定性分析.结果表明,三疣梭子蟹精巢属于叶型,由精小叶和精小管组成的叶状管交织而成.精子发生过程与其他高等短尾派蟹类相似,精子成熟后即进入精小管中.输精管可能起源于精小管.三疣梭子蟹精巢和输精管的发育存在一定的年际变化.4-5月份,精巢小叶内精原细胞占优势,输精管内精荚零星分布;6-7月份,精母细胞与精细胞比例增加;8月份精母细胞为主.到了9月份,整个精巢完全被精细胞等发育程度较高的精小叶所占据,输精管内充满精液和精荚,为即将到来的交配季节做好准备.三疣梭子蟹交配活动主要集中在10月份.12月份精巢精小叶中出现游离精子.此后,精巢逐渐退化(1-3月),但输精管中一直储存着精液和精荚.三疣梭子蟹精荚形成过程始于前输精细管,精荚膜两层,分别由来自前输精细管分泌的第一种不定物质和其中、后端分泌的第二种不定物质.精荚内的基质可能直接源于精巢的精小叶部分.输精管中的强嗜酸性颗粒与第二种不定物质有融合作用,使输精管中贮存的精荚膜嗜酸性.     

栉孔扇贝（♀）×虾夷扇贝（♂）精子入卵过程的电镜观察

采用扫描电镜和透射电镜技术对栉孔扇贝(Chlamys farreri,)×虾夷扇贝(Patinopecten yessoensis,)的精子入卵过程进行观察。结果表明,这2种扇贝杂交与亲本自交的精子入卵过程没有本质的区别。成熟的精卵相遇时,相互激活,产生一系列胞间反应,卵子的激活集中表现为皮层反应、受精膜举起、成熟分裂重新启动;精子激活集中表现为顶体反应和受精锥形成。在16～18℃条件下,精子入卵的时间集中在精卵混合后的第6 min。8 min后,精子的线粒体随精核一起进入卵子中。精子入卵后精核略微膨胀,卵细胞中的线粒体在精核附近出现聚集现象。杂交中有多精入卵现象。本研究目的旨为优化扇贝种间杂交技术及探讨种间受精生物学过程提供基础依据。     

Motility Parameters of Rainbow Trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) Spermatozoa in Relation to Sequential Collection of Milt,Time of <Emphasis Type="Italic">Post-mortem</Emphasis> Storage and Anesthesia

The present study investigated the effects of sequential collection of milt, time of post-mortem storage and anesthesia on rainbow trout (Oncorhynchus mykiss) sperm motility parameters (using computer-assisted sperm analysis – CASA) as well as seminal plasma osmolality and sperm concentration. The post-mortem storage and time of anesthesia altered motility characteristics of rainbow trout sperm to different extents. The moderate impact of time of anesthesia was manifested in a shortened duration of sperm motility after 10 min exposure of fish to anesthetic. The prolonged post-mortem storage (≥40–60 min), in addition to lowering sperm motility duration, also significantly influenced sperm motility parameters, such as sperm velocities, percentage of motile sperm and sperm trajectory parameters. These results clearly demonstrate that when milt from sacrificed fish is used for sperm motility studies, the time of post-mortem storage significantly alters sperm motility characteristics. Since sperm motility rate and swimming velocity could predict fertilizing ability, detrimental effects of prolonged post-mortem storage may lead to reduced fertilization success. Sperm concentration and seminal plasma osmolality were lower in the first fractions and increased with successive collections of milt. It suggests the presence of urine contamination of the first milt fractions which were collected by stripping. Therefore, testing of sperm concentration and/or seminal plasma osmolality should be mandatory while handling stored milt.     

Relationship between biochemical constituents of fish semen and fertility: the effect of short-term storage

Spermatozoa and seminal plasma obtained from rainbow trout and whitefish were analyzed in respect to their aspartate aminotransferase (AspAT) and alkaline phosphatase activities. In particular, the experiments characterized AspAT optimum pH, optimization of assay conditions and action of coenzyme, pyridoxal 5-phosphate (vitamin B₆). The effect of short-term semen storage at 0°C on biochemical indicators and fertilization rate was examined in both species. The concentrations of reduced and oxidized ascorbic acid in seminal plasma of both species were several folds higher than in spermatozoa and blood plasma of fish. Highly significant correlations were found for both species between AspAT activity (sperm or seminal plasma) and fertilization rate (% of eyed-stage or hatched embryos). For rainbow trout, highly significant correlations were found between sperm concentration, motility and fertilization rate. These results suggest that several biochemical indicators of seminal plasma can be used as measures of sperm quality of fish. Some common biochemical parameters for fish and mammal's semen provide evidence for using fish sperm as a model in biomedical research.     

Quality of testicular semen of the African catfish Clarias gariepinus (Burchell, 1822) and its relationship with fertilization and hatching success

Quality differences of testicular semen of the African catfish, Clarias gariepinus, and their influence on fertilization and hatching success were investigated. In accordance with an earlier study, two semen types of the African catfish were distinguished according to testicular maturity stage. Semen type I derived from males with white mature testes whereas type II semen derived from males with grey, partly mature testes. Semen volume, sperm cell concentration and seminal plasma pH was significantly higher in type I semen than in type II semen, while sperm motility was similar. Similar fertilization percentages were obtained with semen type I and semen type II. However, the hatching percentage was higher and the percentage of deformed hatched larvae was lower for type I semen. There were significant (P<0.01) positive correlations between sperm motility and fertilization percentage, seminal plasma pH and hatching percentage and a negative correlation between seminal plasma pH and percentage of deformed larvae. Therefore seminal plasma pH and sperm motility are useful to predict semen quality of the African catfish.     

拟目乌贼繁殖行为学的初步研究

拟目乌贼在繁殖期间的行为较为复杂,包括求偶、雄性争斗、交配、雄性陪护、产卵等行为。拟目乌贼在生殖策略上并非"一夫一妻"制,交配时,雄性用第1、2对腕固定雌性的头部,然后用茎化腕将精荚输送至雌性纳精囊内。雌性受精后不久便产卵,拟目乌贼雌性轮流在产卵区产卵。卵白色,半透明,卵群葡萄状,分枝,刚产出的卵短径约1.2cm,长径约2.4cm,拟目乌贼平均产卵量为354枚。     

羊驼人工采精及授精技术的研究和应用

羊驼常用的人工采精方法主要是假阴道法采精和电刺激法采精。其精液特性不同于其他家畜及野生动物,主要表现为精液量少、精子密度低及精液呈高粘性,这些因素阻碍了羊驼人工采精及授精技术的发展。采精的精液用于授精时要考虑诱导排卵的方法和时间、精液质量及输精时间等因素。重点讨论了羊驼人工采精和授精技术的研究进展及其应用。     

脉红螺交配后性选择机制

交配后性选择是雌性混交动物中的一种普遍现象,其选择机制较为复杂,是性选择研究领域中的重点和难点问题.脉红螺是我国重要的大型经济贝类之一,繁殖形式为多雌多雄混交模式.为了研究该模式繁殖机制,本研究利用6个微卫星标记及行为学观察对脉红螺3个家系进行父权分析.结果表明,3个家系均存在多父性现象,证实脉红螺繁殖属于真正遗传意义上的雌性混交模式.3个家系中父权比例与父本的6个形态学参数及交配持续时间不存在关联性,但均表现出最先与母本交配的父本其后代比例最高,而最后交配的父本其后代比例最低,表明脉红螺交配后性选择存在最先雄性精子优先受精现象,这一现象符合“topping off”假说,即最先交配的雄性排放了大量精子占据了受精囊的主要空间,导致其子代的比例最高;而最后交配的雄性由于受精囊空间的限制,其精子占据空间最少.本研究为海洋无脊椎动物的交配后性选择机制研究提供重要参考资料.     

The antioxidant system of seminal fluid during in vitro storage of sterlet <Emphasis Type="Italic">Acipenser ruthenus</Emphasis> sperm

The role of the seminal fluid antioxidant system in protection against damage to spermatozoa during in vitro sperm storage is unclear. This study investigated the effect of in vitro storage of sterlet Acipenser ruthenus spermatozoa together with seminal fluid for 36 h at 4 °C on spermatozoon motility rate and curvilinear velocity, thiobarbituric acid reactive substance level, and components of enzyme and non-enzyme antioxidant system (superoxide dismutase and catalase activity and uric acid concentration) in seminal fluid. Spermatozoon motility parameters after sperm storage were significantly decreased, while the level of thiobarbituric acid reactive substances, activity of superoxide dismutase and catalase, and uric acid concentration did not change. Our findings suggest that the antioxidant system of sterlet seminal fluid is effective in preventing oxidative stress during short-term sperm storage and prompt future investigations of changes in spermatozoon homeostasis and in spermatozoon plasma membrane structure which are other possible reasons of spermatozoon motility deterioration upon sperm storage.     

Characterization of rainbow trout milt collected with a catheter: semen parameters and cryopreservation success

Collection of fish milt by stripping risks the danger of milt contamination by urine. This may seriously influence milt characteristics and quality, including usefulness for cryopreservation. Urine contamination of milt may be avoided by using a catheter for sperm collection. The objectives of this study were to provide basic characteristics of milt collected with a catheter, to test the usefulness of this milt for cryopreservation, and to correlate characteristics of fresh and cryopreserved semen with sperm fertility rates. Milt from 25 rainbow trout Oncorhynchus mykiss (Walbaum) males were used. All samples were cryopreserved using the pellet method within 1 h of collection, using 0.6 m sucrose and 10% dimethyl sulphoxide (DMSO) as an extender. Catheterization resulted in semen of very good motility (> 90% motile spermatozoa) and high fertilization rates after cryopreservation (mean fertilization rate 81.8 ± 13.3% of control, at a sperm/egg ratio of 2.4 ± 0.3 × 10⁶). Osmolality of seminal plasma and concentrations of sodium, potassium and magnesium ions had low variability, which suggests that they are important for creating a stable environment for sperm storage in the sperm duct. Higher variability of certain seminal plasma characteristics, such as protein concentration and antiproteinase activity, suggests that these characteristics are related to individual semen features of particular males. A strong correlation of seminal plasma zinc concentration with protein concentration may reflect an importance of zinc in semen biology. Cryopreservation caused a significant release of protein and acid phosphatase from spermatozoa. Our results did not reveal any single characteristic of semen collected by catheter that could be used as a powerful predictor of cryopreservation success, presumably because all samples were of high quality.     

中华绒螯蟹雄性生殖系统生化组成及精子代谢

通过对中华绒螯蟹肝胰腺及生殖系统各组织生化成分和乳酸脱氢酶(LDH)活性的测定发现:总碳水化合物的含量以副性腺及输精管内容物(含精荚和精浆)较高,为33.456mg·g-1湿重和21.537mg·g-1湿重,肝胰腺、精荚及精巢的含量较低;碳水化合物中的糖原和葡萄糖含量分别以输精管内容物和肝胰腺最高,分别为0.385mg·g-1湿重和5.866mg·g-1湿重,精巢和肝胰腺中的糖原含量太低而无法检测到,精荚葡萄糖含量最低;蛋白质含量以肝胰腺最高,其余各部分含量显著低于肝胰腺;脂类含量以精巢和肝胰腺最高,其中的甘油三酯仅存于肝胰腺和精巢,且肝胰腺含量显著高于精巢。乳酸脱氢酶活性以贮精囊最高,精荚仅为中等,这可能与精荚壁的通透性有关,而精浆和肝胰腺显著低于其它组织。结果表明,中华绒螯蟹能量代谢主要物质是脂类和蛋白质;精子代谢的能量物质为碳水化合物,并由精浆提供,而其代谢的方式以厌氧代谢为主。