隆线溞生殖细胞成熟分裂过程中染色体的组型观察

取隆线溞[Daphnia (Ctenodaphnia)carinata]孤雌溞的生殖腺细胞进行染色体数量和核型分析,得到隆线溞染色体数目为n=10,2n=20,其核型公式为2n=20=6M+4SM.隆线溞染色体的数目与栉溞亚属[Daphnia(Ctenodaphnia)]中大多数种类一致,进一步验证了隆线溞的分类地位.在此基础上,取隆线溞孤雌溞的卵子(简称夏卵)、两性雌溞的卵子(简称冬卵)及雄溞的生殖腺,利用节肢动物染色体压片的方法,比较观察3种生殖状态的溞体在生殖细胞成熟分裂过程中染色体的组型变化.结果表明:隆线涵孤雌溞的卵子不需交配直接排入孵育囊,只经历1次成熟分裂,然后胚胎开始卵裂.两性雌溞的卵子则在与雄溞交配后排入孵育囊,卵子经历2次正常的成熟分裂,第一次为减数分裂,第二次为有丝分裂,姐妹染色单体分开,形成单倍体的配子,配子与精子结合形成受精卵,然后开始卵裂.隆线溞雄溞生殖细胞同样经历了2次正常的成熟分裂,第一次为减数分裂,第二次为有丝分裂,形成单倍体的精子.隆线溞这种既具有低等生物的孤雌生殖方式,又有高等动物的有性生殖方式,可能是其在长期进化过程中对环境的适应性表现,也可能是动物由低等向高等进化的-个中间过程.本研究通过对隆线溞孤雌溞、两性雌溞及雄溞生殖细胞发生过程中染色体的组型变化观察,初步阐明了隆线溞生殖发育的变化规律,旨在为实现枝角类生殖发育的人为调控奠定基础.     

多刺裸腹溞的精子发生

应用组织学方法和电镜技术,研究了多刺裸腹溞(Moina macrocopa)的精子发生、成熟精子的形态以及雄性生殖系统的结构特征。多刺裸腹溞雄性生殖系统包括1对精巢、1对输精管和1个生殖孔。精巢壁薄,由电子密度不同的2层结缔组织膜组成。生殖孔位于尾爪底部,其周围无类似交媾器等其他附属结构。多刺裸腹溞的精子发生经历精原细胞、精母细胞、精子细胞和成熟精子4个主要时期。精原细胞呈椭圆形,核染色质较分散,细胞质中分布着大量的粗面内质网和游离的核糖体。精母细胞由于挤压呈多角形,细胞核为椭圆形,核基质和染色质分别位于细胞核的一侧,细胞质中存在少量的线粒体,内质网呈片层状。精子细胞向精子分化,形态上纵向拉长,细胞核也纵向伸长;细胞质中线粒体数量增多,个体增大,且部分线粒体内嵴溶解,形成空泡状的衍生物;内质网大量发生。成熟精子弯曲成棒形,细胞核变小,呈圆形,位于精子的中央。扫描电镜观察显示,精子外表面光滑无附属结构。在整个精子发生过程中,细胞核双层核膜结构完整且清晰。此外在精巢中还存在巨大细胞。     

文蛤卵母细胞卵黄发生的超微结构

取不同发育阶段文蛤 (MeretrixmeretrixLinnaeus)活体解剖取出生殖腺 ,常规方法制成切片 ,以透射电镜(TEM )观察其卵母细胞的卵黄发生过程。结果表明 ,文蛤的卵母细胞卵黄发生期间 ,线粒体、内质网、高尔基复合体、溶酶体、微吞饮泡等细胞器均参与了卵黄粒的形成。卵黄合成早期的卵母细胞质膜伸出大量的微绒毛 ,并出现卵黄膜 ,胞质中有大量膜性小泡 ,溶酶体、线粒体、高尔基复合体和粗面内质网发达 ,卵黄前体物质逐渐增多。在卵黄合成中期 ,胞质内线粒体和内质网活动活跃 ,卵母细胞大量合成和积累卵黄物质 ,细胞质膜外凸 ,与外界进行物质交换。卵黄合成后期 ,卵质内贮存了大量的卵黄粒 ,细胞器不发达。此外 ,还对卵母细胞发育过程卵黄颗粒的细胞内、外原料来源进行了讨论     

秀丽白虾卵母细胞不同发育阶段线粒体的变化

采用透射电镜技术观察秀丽白虾(Exopalaemon modestus)卵巢不同发育阶段卵母细胞线粒体的变化及其与卵黄发生的关系，结果显示，随着卵母细胞的发育，线粒体经历了数目以及结构上的系列变化。卵黄发生早期，卵母细胞内的线粒体数量急剧增多，形态各异，有椭圆形、圆球形等，线粒体嵴稀少；随后，细胞质中出现较多近椭圆形的线粒体群，嵴丰富，外膜光滑。连续切片观察表明，该时期线粒体尚未明显参与卵黄颗粒的形成。卵黄发生中期，卵母细胞内的线粒体的结构出现两大类群：一类线粒体呈长杆状或椭圆形，体积小，嵴丰富，内部基质暗，外膜光滑，主要功能是为卵母细胞发育提供能量；另一类线粒体体积大，呈圆球型，嵴逐渐退化，内部基质呈现电子透明状，外膜出现大小不一的波浪状。后一类线粒体膜内陷，出现明显的类似细胞内吞作用，参与卵黄颗粒的形成。被吞饮物主要为有膜性泡，包括波浪状膜的高尔基体分泌泡、光滑外膜的内质网囊泡以及小型线粒体。与此同时，卵母细胞质膜形成众多胞饮小泡，外来物质逐渐进入卵母细胞，通过高尔基体的加工，以分泌泡相互融合形式参与卵黄颗粒的形成，内吞作用所累积的卵黄颗粒占主要部分，因此在该时期卵母细胞卵黄颗粒的形成以外源性为主。在卵子发育后期，皮层区域出现具典型结构的线粒体，当这些线粒体参与皮层颗粒形成后，卵子逐渐成熟，整个卵母细胞中很难见到线粒体。卵母细胞内卵黄颗粒进一步形成并相互融合，整个卵细胞几乎全为卵黄颗粒占据，未见明显线粒体。     

四川华鳊卵子发生的显微结构观察

为探究四川华鳊(Sinibrama taeniatus)卵子的发生规律,对其卵子发生进行了显微结构观察和描述。将卵子发生分为卵原细胞(Ⅰ时相)、单层滤泡时相(Ⅱ时相)、卵黄泡出现时相(Ⅲ时相)、卵黄充满时相(Ⅳ时相)和成熟卵子(Ⅴ时相) 5个时相。观察发现,四川华鳊卵原细胞存在2种形态,分别为早期卵原细胞和有丝分裂过程中的卵原细胞;同时将早期初级卵母细胞划入Ⅱ时相,此阶段细胞核膜边缘出现多个小核仁,将其作为小生长期开始的标志;Ⅲ时相卵黄泡出现,当卵黄泡积累至3～5层时,卵黄物质即开始积累,此时多以卵黄颗粒的形式存在;进入Ⅳ时相后,卵黄物质沉积形成卵黄小板,卵黄泡被挤压至卵周,形成皮层小泡,卵子成熟后,原皮层小泡所在区域呈块状或颗粒状,经苏木精-伊红(HE)染色后呈橘红色。     

中华绒螯蟹卵黄发生期卵母细胞和卵泡细胞超微结构观察

通过透射电镜技术观察了中华绒螯蟹第二次卵巢发育过程中卵巢的超微结构变化.结果表明:(1)中华绒螯蟹第二次卵巢发育过程中卵黄发生期可分为初期和后期;(2)卵黄发生初期(雌蟹第一次排卵后的16 d内),卵黄生成以卵母细胞内源性合成为主,此时卵母细胞胞质中存在大量内质网囊泡、高尔基体和线粒体,这些细胞器参与胞内卵黄物质的合成.内源性合成后期,卵母细胞膜形态多样,呈现触手状、波浪状和断裂状,为外源合成期做准备.此期卵泡细胞还未向卵母细胞靠近,两类细胞间存在着由淋巴细胞吐出的絮状物;(3)卵黄发生后期,首先为卵泡细胞与卵母细胞的结合阶段(排卵后16～21 d),此后,卵泡细胞胞质中含有大量内质网囊泡、卵黄颗粒和脂滴,卵母细胞与卵泡细胞膜变为链珠状便于物质交换,卵母细胞的卵黄合成能力减少,转由卵泡细胞进行外源性物质吸收和卵黄物质合成(21～36 d);(4)卵黄发生结束后,双层卵膜形成,卵黄体和脂肪滴均匀分布在卵母细胞胞质中.     

黄海高眼鲽卵巢发育特征及卵径分布

本研究通过组织学观察,描述了黄海高眼鲽(Cleisthenes herzensteini)卵母细胞发育特征及其退化过程。高眼鲽卵母细胞发育分为5个时相:第1时相为卵原细胞,细胞体积小,细胞质少,细胞核明显;第2时相卵母细胞细胞核附近出现卵黄核;第3时相由胞质外缘向内层逐渐产生液泡并生成卵黄颗粒,出现双层滤泡膜;第4时相卵母细胞内充满卵黄,细胞核向动物极移动,放射膜增厚;第5时相细胞核溶解,卵母细胞从滤泡膜中释放出来并发生水合作用;产卵期过后,卵巢发生退化,卵黄颗粒逐渐被吞噬,放射膜溶解断裂。通过比较卵巢中各时相卵母细胞组成比例,表明卵母细胞发育具有非同步性。Ⅳ、Ⅴ、Ⅵ-Ⅳ'期卵巢内卵径(长径)呈单峰分布,优势粒径组分别为0.45~0.55 mm、0.60~0.65 mm和0.40~0.50 mm;Ⅴ'期卵巢,卵径分布呈双峰型,峰值分别为0.50~0.55 mm和0.90~0.95 mm,水合卵母细胞与卵径较小的小生长期卵母细胞比例增大,呈现出明显的双峰分批产卵型特征。     

中华绒螯蟹卵黄发生期卵母细胞和卵泡细胞超微结构观

通过透射电镜技术观察了中华绒螯蟹第二次卵巢发育过程中卵巢的超微结构变化。结果表明：（1）中华绒螯蟹第二次卵巢发育过程中卵黄发生期可分为初期和后期;（2）卵黄发生初期（雌蟹第一次排卵后的16 d内）,卵黄生成以卵母细胞内源性合成为主,此时卵母细胞胞质中存在大量内质网囊泡、高尔基体和线粒体,这些细胞器参与胞内卵黄物质的合成。内源性合成后期,卵母细胞膜形态多样,呈现触手状、波浪状和断裂状,为外源合成期做准备。此期卵泡细胞还未向卵母细胞靠近,两类细胞间存在着由淋巴细胞吐出的絮状物;（3）卵黄发生后期,首先为卵泡细胞与卵母细胞的结合阶段（排卵后16~21 d）,此后,卵泡细胞胞质中含有大量内质网囊泡、卵黄颗粒和脂滴,卵母细胞与卵泡细胞膜变为链珠状便于物质交换,卵母细胞的卵黄合成能力减少,转由卵泡细胞进行外源性物质吸收和卵黄物质合成（21~36 d）;（4）卵黄发生结束后,双层卵膜形成,卵黄体和脂肪滴均匀分布在卵母细胞胞质中。     

锈斑蟳雌性生殖系统的组织学研究

试验结果表明,锈斑蟳雌性生殖系统由卵巢、输卵管、生殖孔和受精囊组成.卵巢由外膜和内生殖上皮构成;输卵管较短,由外膜、肌层和上皮构成;生殖孔位于输卵管末端与受精囊相连;受精囊由结缔组织、薄肌层和上皮组织构成.卵子的发生可根据卵细胞及核仁的大小、形态和卵黄积累分为:卵原细胞期、初级卵母细胞期、卵黄合成初期卵母细胞、卵黄合成中期卵母细胞、卵黄合成后期卵母细胞、卵母细胞发育晚期、卵母细胞成熟期.     

翎电鳗卵巢发育组织学研究

翎电鳗是深受广大水族爱好者喜爱的观赏鱼类,研究其卵巢发育规律,可以为该鱼的人工繁育提供理论基础。翎电鳗成熟卵巢为囊状,泄殖孔位于体前端下颌后方。常规石蜡切片方法对其卵巢进行观察发现,翎电鳗的卵巢发育分为6个时期,卵子发育分为5个时相。4月龄雌鱼卵巢中有较多第Ⅱ时相卵母细胞,少量卵原细胞,发育至第Ⅱ期;8月龄雌鱼卵巢中含有卵原细胞、第Ⅱ时相和第Ⅲ时相卵母细胞,发育至第Ⅲ期;10月龄雌鱼卵巢中同时存在卵原细胞、第Ⅱ时相、第Ⅲ时相和第Ⅳ时相卵母细胞,发育至第Ⅳ期,达性成熟;11月龄雌鱼卵巢中第Ⅴ时相卵母细胞占主要成分,为Ⅴ期。Ⅴ期卵巢中成熟卵径的大小分布在1.08～1.15 mm和1.78～1.84 mm。翎电鳗雌性亲鱼在理想环境下存在短时间内一批卵排出后新一批次的卵成熟并被释放的可能,产卵类型属分批同步型。     

Ovarian maturation stages of the mud crab Scylla serrata

Ovarian maturation in adult wild‐sourced and pond‐grown Scylla serrata (Forsskål) was determined based on gross morphology and histological appearance. There were no significant differences noted in the histological features of both wild and pond‐reared S. serrata females. Ovarian maturation was classified into five stages: immature, early maturing, late maturing, fully mature and spent. The immature ovaries are thin and translucent to off white and contain oogonia, primary oocytes with large nuclei. The follicle cells were found around the periphery of the lobes and an area among groups of oogonia and oocytes. The follicle cells gradually enclosed the oocytes. The early‐maturing ovaries were yellow and small yolk globules started to appear in larger oocytes. In late‐maturing ovaries, the colour became light orange and lobules were apparent. Yolk globules occurred in the cytoplasm with larger globular inclusions towards the periphery, while follicle cells were hardly recognizable. Fully mature ovaries were orange to deep orange and had swollen lobules. Large yolk globules were apparent in the entire cytoplasm. Follicle cells were hardly seen. Spent ovaries were similar to the early‐maturing and late‐maturing stage in partially spawned females. The ovarian development was correlated closely to the gonadosomatic index, oocyte diameter, and ovarian histology. The classification of ovarian maturation provides baseline information for further studies on reproductive biology. Likewise, the information provides a guide for broodstock management in the hatchery.     

池养鲻的卵巢发育和卵子发生过程

通过卵巢切片的组织学分析揭示，幼鲻在土池养殖3个月后可见线状卵巢，大约5个月后卵原细胞进入第一次成熟分裂前期的双线期转变为早期初级卵母细胞。接着卵母细胞生发泡(核)和胞质体积增加，核质比从3．5：1减少至2：1。此后卵巢中卵母细胞停滞发育持续至养殖的第3年。在第3年卵巢切片看出卵母细胞进入脂肪泡时相，在第3年秋季进入卵黄发生时相。但在人工养殖条件下卵母细胞仅能发育到卵黄发生后期，即卵母细胞胞质充满卵黄颗粒，生发泡居中而不移位。这些结果对于用人工养殖鲻为亲鱼开展人工繁殖提供重要的科学依据。并讨论了卵子发生6个时相的生物学特点及其重要的细胞器在卵黄发生中可能的生理作用。     

凡纳滨对虾卵子发生的超微结构

利用透射电镜研究凡纳滨对虾卵子发生过程中细胞内部结构的变化。结果显示,卵原细胞结构简单,代谢水平低,核孔稀少,通过核周池来完成核、质之间的物质交换。卵黄发生前晚期和卵黄发生初期的卵母细胞变化显著：核膜凹凸不平,核仁数量多,核孔密集,大量核仁外排物经核孔输送到卵质中;卵质中胞器极为丰富和发达,代谢活性极强。卵黄发生旺盛期是卵黄大量形成的阶段,卵质边缘还呈辐射状排列了一圈椭圆形皮质棒,细胞出现微吞饮活动并形成卵黄膜。卵黄发生晚期卵质中充满了粗大的卵黄粒和脂滴,胞器锐减。另外,探讨了卵细胞内部结构的变化和卯黄形成的关系以及皮质棒的来源与功能。     

Oogenesis in the common Japanese conger Conger myriaster

ABSTRACT:    We investigated the process and characteristics of oogenesis in the common Japanese conger Conger myriaster . Young fish caught in November 1996 were reared for use in this experiment. Fish were sampled monthly from December 1997 to August 1998. Some were injected with human chorionic gonadotropin to stimulate ovarian maturation from May to August 1998. Oocytes from the chromatin nucleolus stage to the secondary yolk globule stage were obtained from non-hormone-treated fish; those of more advanced stages were obtained from hormone-treated ones. We divided oocyte development into eight stages from the chromatin nucleolus stage to the maturation stage. The yolk vesicle stage was not separated because yolk vesicles began to appear just after appearance of yolk globules. Oocyte, oil droplet, yolk globule and nucleus diameters all increased concomitant with oocyte development. Oil droplet and yolk globule diameters increased remarkably at the maturation stage. However, zona radiata thickness peaked at the secondary yolk globule stage, decreasing gradually thereafter. Increased gonadosomatic index was related to oocyte development as found in European and Japanese eels receiving hormone treatment to mature. The present study is the first report describing oogenesis characteristics in congrid eels. It indicates that oogenesis is almost identical to that of other anguillid eels.     

孔雀鱼雌性生殖器官组织结构及卵子发生的研究

应用光镜技术研究了孔雀鱼（Poecilia reticulata）雌性生殖器官组织结构和卵子发生的特点,以期为孔雀鱼的良种繁育工作提供基础资料。结果表明,孔雀鱼雌性生殖器官由卵巢、卵子输送管和泄殖孔构成。卵子发生过程经历了6个时相：第Ⅰ时相主要为卵原细胞;第Ⅱ时相是处于小生长期的初级卵母细胞,形成生长环、卵黄核和单层扁平滤泡细胞膜;第Ⅲ时相是进入大生长期的初级卵母细胞,卵黄开始积累,形成2层滤泡细胞膜和2层卵包膜;第Ⅳ时相为发育晚期的初级卵母细胞,细胞发生了极化,卵包膜发育完善;第Ⅴ时相发育成熟,卵黄颗粒凝结呈块状,卵黄泡主要分布于细胞膜内缘,滤泡细胞膜萎缩;第Ⅵ时相是闭锁卵泡,被滤泡细胞消化和吸收转变为卵巢内结缔组织。     

太平洋牡蛎二倍体和三倍体卵母细胞发育的超微结构

本文利用电子显微镜比较研究了太平洋牡蛎二倍体和三倍体卵母细胞发育过程中细胞和各细胞器的超微结构变化。结果表明,在卵黄形成期,二倍体卵母细胞呈椭圆形,细胞器发达,细胞外布满微绒毛,卵黄粒多并均匀分布于细胞质中,卵母细胞生物合成旺盛,代谢活动强;而三倍体卵母细胞呈长条形或不规则形,细胞器少,不发达,细胞外未观察到微绒毛,卵黄粒少,有些细胞的卵黄粒畸形,卵母细胞生物合成及代谢弱。     

Classification of differentiating oocytes during ovarian cycle in the giant freshwater prawn, Macrobrachium rosenbergii de man

Based on the light microscopic observations of cells' sizes, chromatin patterns, amount of lipid droplets and yolk granules, the female germ cells could be classified into four different phases, which include 1) oogonia (Oog), 2) primary oocytes (pOc), 3) secondary oocytes (sOc), and 4) mature oocyte (mOc). Oog are small oval-shaped cells with irregular-shaped nuclei sizing 4–6 μm in diameter. They rest on the connective tissue germinal cord at the tip of each ovarian pouch (lobule). Oogonia increase their number through mitotic division, and the daughter cells move into ovarian pouch where they undergo first meiotic division to become primary oocytes, which have various steps of 1st meiotic prophase accumulating at the innermost zone of the ovarian pouch. The primary oocytes are small oval-shaped cells (8.5–10 μm in diameter) with large nuclei containing chromatin in various states of condensation that finally transform into chromatids. Their nuclei are surrounded by thin rim of faint blue-stained cytoplasm. The secondary oocytes derived from 2nd meiosis and comprise five steps: Oc1 and Oc2, classified as previtellogenic oocytes, Oc3 and Oc4, classified as vitellogenic oocytes, and mature oocyte (mOc) The zones of ovarian pouch are defined based on the accumulation of various steps of developing oocytes, namely, oogenic, previtellogenic, vitellogenic and mature zones, respectively. The ovarian cycle is divided into five stages based on the number and types of oocytes present in each stage. Stage 0 and I are spawn and spent stages. Stage II and III are proliferative and premature stages, while stage IV is mature stage. During ovarian stage I, each ovarian pouch contains primarily oogonia, primary oocytes, Oc1 and a few Oc2. In stage II, the pouch contains mainly Oc2 and Oc3, while in stage III the predominant cells are Oc4. Mature oocytes appear synchronously, in stage IV. The ovulating mature oocytes pass through the thin disrupted wall of ovarian pouch into subcapsular space, that leads into the oviduct situated on the ventro-lateral side of the ovarian lobe. At spawning, the ovarian pouches break down and only connective sheaths and hemolymph sinuses remain. The germinal cords and islets of oogonia remain in the central area of stage 0 ovary. The ovarian capsule, including the muscular layer, becomes attenuated as the ovary progresses from stage 0 to IV. The hemolymph vessels become highly convoluted in the central area of the ovary, and they branch radially into smaller hemolymph sinuses around each oogenic pouch.     

The reproductive biology of the amberjack, Seriola dumerilii (Risso 1810). I. Oocyte development in captivity

A study of the reproductive biology of the amberjack, Seriola dumerilii, held in captivity was carried out, describing oogenesis as well as the different stages of the ovarian cycle. Seven stages of oocyte development, as well as oogonia, were distinguished. Cortical alveoli were hardly detectable within the oocyte, as they were small, sparse and contained few mucopolysaccharides. It is suggested that their role in the fertilization process might be less important than in other teleost species. Fish aged 3 and 4 years were found to be sexually immature, with ovaries containing only previtellogenic oocytes. Vitellogenesis started in December in fish aged 4 + years. Late-developing ovaries showing deposition of yolk protein granules were found at the end of the 5th year of life (May) in specimens measuring 80.0 ± 3.5 cm standard length. This should be regarded as the minimum size at which sexual maturity is reached in S. dumerilii. Final vitellogenesis and oocyte maturation were, however, inhibited in captivity, and extensive follicular atresia took place as the natural spawning season approached. It is suggested that insufficient gonadotrophic stimulation because of confinement stress may be the cause of failed maturation and spawning in this species under culture conditions.