莫桑比克非鲫卵黄形成的电镜观察

运用透射电镜观察了莫桑比克非鲫卵母细胞的生长.根据卵母细胞的大小和内部结构特征,将其分为四个时期:卵母细胞生长早期:卵黄泡形成期:卵黄积累期:卵黄积累完成期.本文着重研究了主要卵黄成分--卵黄球的形成过程.卵黄球属外源性卵黄,由卵母细胞通过微胞饮作用吸收肝脏合成的卵黄蛋白原后形成的.在卵黄大量积累前,卵母细胞内的线粒体和多泡体聚集成团,构成卵黄核,继而线粒体大量增殖,线粒体形状发生改变,形成同心多层膜结构,为大量的卵黄物质积累提供场所.最终形成的卵黄球由被膜、卵黄结晶体和两者之间的非结晶区三部分组成.       

黄胫小车蝗卵子发生及卵母细胞凋亡的显微观察

对黄胫小车蝗(Oedaleus infernalis)卵子发生过程和卵母细胞凋亡进行显微观察。结果表明,黄胫小车蝗卵子发生可明显分为3个时期10个阶段,即卵黄发生前期、卵黄发生期和卵壳形成期。第1阶段,卵母细胞位于卵原区,经历减数第一次分裂;第2阶段,卵母细胞核内染色体解体成网状,滤泡细胞稀疏地排列在卵母细胞周围;第3阶段,滤泡细胞扁平状,在卵母细胞周围排成一层;第4阶段,滤泡细胞呈立方形排在卵母细胞周围;第5阶段,滤泡细胞呈长柱形排在卵母细胞周围,滤泡细胞之间、滤泡细胞与卵母细胞之间出现空隙;第6阶段,卵母细胞边缘开始出现卵黄颗粒;第7阶段,卵母细胞中沉积大量卵黄,胚泡破裂;第8阶段,滤泡细胞分泌卵黄膜包围卵黄物质;第9阶段,滤泡细胞分泌卵壳;第10阶段,卵壳分泌结束,卵子发育成熟。卵母细胞发育过程中的凋亡发生在卵黄发生前期,主要表现为滤泡细胞向卵母细胞内折叠,胞质呈团块状等特征。     

北京油葫芦卵子发生中DNA及RNA动态变化研究

用孚尔根及甲基绿 -派洛宁组织化学染色法了解北京油葫芦Teleogryllusmitratus(Burmeister)卵子发生各时期阶段中卵内DNA及RNA动态变化规律。在卵子发生的最初阶段 ,核中DNA的合成和复制最活跃 ,以后便慢慢减弱 ;而RNA则在第 2阶段合成最旺盛。在卵子发生各个阶段 ,滤泡细胞中DNA ,RNA均为阳性反应 ,并在卵细胞的卵黄形成期活动旺盛 ,为卵母细胞卵黄蛋白形成提供物质基础。卵子发生第 4～ 6阶段 ,滤泡细胞开放时期 ,血淋巴内一些物质可能直接或间接通过滤泡细胞间隙进入卵母细胞内 ,参与卵母细胞的发育和构建。研究表明卵子发生初期卵母细胞的发育和物质构建主要以内源性合成积累为主 ,中后期则有外源性物质的参与。     

泥螺卵子发生的超微结构研究

利用透射电镜观察了泥螺卵子发生过程。结果表明 ,泥螺的卵子发生可划分为卵原细胞、卵黄发生早期、卵黄发生中期及卵黄发生后期卵母细胞 4个时期。卵原细胞核大而圆 ,胞质内分布有少量的线粒体和高尔基囊泡 ,细胞表面具微绒毛。卵黄发生早期的卵母细胞 ,胞质中各类细胞器发达 ,并出现数量较多的类朦胧子。卵黄发生中期的卵母细胞胞体迅速增大 ,核伸出伪足状突起 ,卵质中各种细胞器活动活跃 ,并参与形成卵黄粒和脂滴。此期还可观察到卵母细胞与滤泡细胞间的物质交换现象。卵黄发生后期的卵母细胞体积增至最大 ,细胞器数量减少。本文就卵黄发生前后卵母细胞内部构造的变化、意义及滤泡细胞与卵母细胞蛋白来源间的关系作了探讨     

泥螺卵子发生的超微结构研究

利用透射电镜观察了泥螺卵子发生过程。结果表明,泥螺的卵子发生可划分为卵原细胞、卵黄发生早期、卵黄发生中期及卵黄发生后期卵母细胞4个时期。卵原细胞核大而圆,胞质内分布有少量的线粒体和高尔基囊泡,细胞表面具微绒毛。卵黄发生早期的卵母细胞,胞质中各类细胞器发达,并出现数量较多的类朦子。卵黄发生中期的卵母细胞胞体迅速增大,核伸出伪足状突出,卵质中各种细胞器活动活跃,并参与形成卵黄粒和脂滴。此期还可观察到卵母细胞与滤泡细胞间的物质交换现象。卵黄发生后期的卵母细胞体积增至最大,细胞器数量减少。本文就卵黄发生前后卵母细胞内部构造的变化、意义及滤泡细胞与卵母细胞蛋白来源间的关系作了探讨。     

山溪鲵卵黄发生的显微与超微结构

用光镜和透射电镜观察了山溪鲵（Batrachuperus pinchonii)不同发育时期卵母细胞的显微与超微结构,特别注意了与卵黄发生相关的细胞器变化。结果表明,类核周体为线粒体、高尔基体、内质网等膜性细胞器聚集的场所;线粒体自身也可演变成卵黄前颗粒,参与构成卵黄小板的成分;吞饮泡和髓样小体是卵母细胞利用外源卵黄物质的中间媒介。经过与其他动物卵黄发生过程相比较,认为非哺乳脊椎动物卵黄发生是卵母细胞在多种细胞器参与下整体活动的结果,不是经由单一的模式或途径形成,因此呈现发生上的多元化;不同物种在卵黄发生中分别采取与各自相适应的模式或途径。     

七星瓢虫卵子发生的观察

对七星瓢虫(Coccinella septempunctata L.)卵子发生过程进行了组织学、细胞学观察及阶段划分,并与取食人工饲料的瓢虫进行对比。卵母细胞仅出现在幼虫期。蛹期已分化为卵母细胞与营养细胞。成虫期卵子发生可以明显的分为卵母细胞分化、卵母细胞营养及卵母细胞卵黄形成三个时期,并分为9个阶段。第1阶段:卵母细胞位于卵原区,进行第一次减数分裂的前期。第2阶段:卵母细胞位于颈区,开始增大,出现了营养索,DNA呈明显的孚尔根正反应。第3阶段:卵母细胞形成卵泡囊并进入生长区,核增大成胚泡。第4阶段:胚泡移至卵质周缘,卵质中RNA丰富,滤泡细胞立方形。第5阶段:胚泡内核仁增大、分枝并释放核仁小体进入卵质。第6阶段:营养索消失,滤泡细胞扁平并出现空位,卵黄形成开始。第7阶段:卵黄球形成逐渐充满卵质,胚泡膜逐渐消失。第8阶段:胚泡消失,滤泡细胞开始分泌卵壳。第9阶段:卵发育完成,经过上皮塞进入输卵管。取食人工饲料瓢虫的卵子发生过程显著缓慢,发育中的卵母细胞致量少,滤泡细胞及卵黄分布均不正常。     

九孔鲍卵子发生及卵巢发育的组织学观察

采用组织学方法研究了九孔鲍(Haliotis diversicolor supertexta)的卵子发生、卵巢结构及其发育.根据卵细胞的大小、形状,核仁的形态,卵黄颗粒的积累情况,滤泡的结构等.将九孔鲍卵子的发生分为卵原细胞、卵黄发生前的卵母细胞和卵黄发生期的卵母细胞3个时期;卵巢壁由外膜及内生殖上皮构成,生殖上皮分化产生卵原细胞和滤泡细胞;卵巢的结构单位是滤泡.根据卵巢的外部形态和内部组织结构,将九孔鲍的卵巢发育分为休止期、增殖期、生长期、成熟期和排放期共5期.     

抑卵激素对家蝇卵巢周期性发育的调控

抑卵激素是调控家蝇Musca dorncstica vicina卵巢周期性发育的关键因子之一。在家蝇中,当第一个周期的卵母细胞处于卵黄发生期或卵黄发生后期时,其第二个周期的卵母细胞的发育不进入卵黄发生期。本文建立了家蝇抑卵激素的生物测定方法,即用一对卵巢提取物注射1头羽化后12h家蝇,并在羽化后60h观察卵母细胞的发育及卵黄蛋白的沉积情况。抑卵激素的作用首先是延缓了卵母细胞在卵黄发生前期的发育;其次,抑卵激素抑制脂肪体中卵黄蛋白的合成,导致血淋巴中卵黄蛋白含量的下降,从而抑制了卵母细胞的发育。抑卵激素并不抑制卵母细胞对卵黄原蛋白的摄取。卵发育神经激素可以颉抗抑卵激素的抑制作用。抑卵激素无种属特异性。     

短额负蝗卵子发生过程中糖复合物的动态分布

以生物素标记的凝集素(UEA-I、SBA、PNA)为探针，利用凝集素组织化学方法对短额负蝗(Atractomorpha sinensis)卵子发生过程中滤泡细胞和卵母细胞内糖复合物的分布进行了定位研究。结果表明，在卵子发生的各期滤泡细胞和卵母细胞中没有UEA-I受体的表达，SBA和PNA受体以不同的分布模式呈阶段性表达。两者首次出现于卵母细胞生长期， 随后PNA受体消失，SBA受体大量表达；在卵黄形成期前期SBA受体和重新出现的PNA受体表达于卵黄颗粒形成部位，卵黄形成期后期两者均为阴性表达；成熟卵子中两种受体又以不同程度重新出现于卵黄膜。两种受体在滤泡细胞内均大量表达 提示，N-乙酰半乳糖胺和半乳糖-β-(1，3)半乳糖胺复合物的修饰和变化与卵母细胞的发育、卵黄物质的形成及滤泡细胞的增殖分化密切相关，卵黄膜上的糖复合物可能与精卵识别有关。     

Ovarian follicle growth in the catfish Iheringichthys labrosus (Siluriformes: Pimelodidae)

The morphofunctional organisation of the female reproductive system, the oocyte growth and the follicular envelope ultrastructure were studied by the first time in the catfish Iheringichthys labrosus from Upper Paraná River basin, Southeastern Brazil, in order to contribute to the knowledge of the reproductive behaviour strategies of this species. As in other Neotropical freshwater siluriforms, the ovaries are of the cystovarian type, the oocytes develop in an asynchronous pattern and mature oocytes are released in clusters in the ovarian lumen, being transported through the oviduct to the urogenital papilla. During the primary growth, nuclear material is transported to the ooplasm, forming the yolk nucleus, where proliferate membranous organelles. The onset of the zona radiata formation occurs during the late perionucleolar stage with the deposition of the outer layer. At the vitellogenic stage, this envelope reaches 6.35+/-0.84microm of thickness, being constituted by three distinct layers crossed by pore-canals containing oocyte and follicular cells processes. Cytochemical analyses evidence neutral glycoproteins in cortical alveoli, yolk globules and zona radiata. Follicular cells with squamous shape during the primary growth acquire synthetic activity at the secondary growth, reaching 37.82+/-4.72mum in height at the mature vitellogenic follicles. These cells accumulate sulphated polysaccharides in large electron-lucent vesicles during the vitellogenic stage which are possibly secreted to form a mucous coat at the egg surface. These evidences suggest that I. labrosus may have adhesive eggs as also detected in other Neotropical freshwater Siluriformes.     

Electron microscopic study of the formation of the egg membranes of the oocytes of the last year of development in the peliad]

After ovulation the egg membrane of the pelled consists of 3 layers. The homogeneous electron-dense layer comprising its major mass zona radiata, is laid as early as the stage of protoplasmatic growth of the oocyte as a result of accumulation of an electron-dense substance of a high electron density in the basement of the oocyte microvilli. The next, pectinate layer is formed from the highly electron-dense substance formed in the vacuoli of the follicular cell Golgi complex released into the subfollicular space. The most external layer - a fibrous layer - is formed due to release of large vacuoles, formed in the apical area of the follicular cells not long before ovulation. The obtained data suggest a great role of the follicular epithelium in the formation of egg membranes of the pelled.     

Oogenesis in the bluefin tuna,Thunnus thynnus L.: a histological and histochemical study

Histology and histochemistry are useful tools to study reproductive mechanisms in fish and they have been applied in this study. In the bluefin tuna, Thunnus thymus L., oocyte development can be divided into 4 principal phases based on the morphological features of developing oocytes and follicles. The primary growth phase includes oogonia and basophilic or previtellogenic oocytes classified as chromatin-nucleolus and perinucleolus stages. The secondary growth phase is represented by vitellogenic oocytes at early (lipid globule and yolk granule 1), mid (yolk granule 2) and late (yolk granule 3) vitellogenesis stages. The maturation phase involves postvitellogenic oocytes undergoing maturation process. During the spawning period, both postovulatory follicles, which indicate spawning, and atretic follicles can be distinguished in the ovary. Carbohydrates, lipids, proteins and specially those rich in tyrosine, tryptophan, cystine, arginine, lysine and cysteine, as well phospholipids and/or glycolipids and neutral glycoproteins were detected in yolk granules. Moreover, affinity for different lectins (ConA, WGA, DBA and UEA) was detected in vitellogenic oocytes (yolk granules, cortical alveoli, follicular layer and zona radiata), indicating the presence of glycoconjugates with different sugar residues (Mannose- Man- and/or Glucose -Glc-; N-acetyl-D-glucosamine- GlcNAc- and/or sialic acid- NANA-; N-acetyl-D-galactosamine- GalNAc-; L-Fucose -Fuc-). Histochemical techniques also demonstrated the presence of neutral lipids in globules (vacuoles in paraffin sections) and neutral and carboxylated mucosubstances in cortical alveoli. By using anti-vitellogenin (VTG) serum, immunohistochemical positive results were demonstrated in yolk granules, granular cytoplasm and follicular cells of vitellogenic oocytes. Calcium was also detected in yolk granules and weakly in follicular envelope. In females, the gonadosomatic index (GSI) increased progressively from May, during early vitellogenesis, until June during mid and late vitellogenesis, where the highest values were reached. Subsequently, throughout the maturation-spawning phases (July), GSI decreased progressively reaching the minimal values during recovering-resting period (October).     

Fine Structure of the Ovarian Development in the Orb-web Spider, Nephila clavata

ABSTRACT Fine structural changes of the ovary and cellular composition of oocyte with respect to ovarian development in the orb-web spider, Nephila clavata were examined by scanning and transmission electron microscopy. Unlike the other arthropods, the ovary of this spider has only two kinds of cells-follicle cells and oocytes. During the ovarian maturation, each oocyte bulges into the body cavity and attaches to surface of the elongated ovarian epithelium through its peculiar short stalk attachments. In the cytoplasm of the developing oocyte two main types of yolk granules, electron-dense proteid yolk and electron-lucent lipid yolk granules, are compactly aggregated with numerous glycogen particles. The cytoplasm of the developing oocyte contains a lot of ribosomes, poorly developed rough endoplasmic reticulum, mitochondria and lipid droplets. These cell organelles, however, gradually degenerate by the later stage of vitellogenesis. During the active vitellogenesis stage, the proteid yolk is very rapidly formed and the oocyte increases in size. However, the micropinocytosis invagination or pinocytotic vesicles can scarcely be recognized, although the microvilli can be found in some space between the oocyte and ovarian epithelium. During the vitellogenesis, the lipid droplets in the cytoplasm of oocytes increase in number, and become abundant in the peripheral cytoplasm close to the stalks. On completion of the yolk formation the vitelline membrane, which is composed of an inner homogeneous electron-lucent component and an outer layer of electron-dense component is formed around the oocyte.     

中华蚱蜢卵子发生中花生凝集素受体的初步鉴定和发育表达

The distributions of PNA binding glycoconjugates in the plasma membrane of Acrida cinerea Thunberg germ cells were detected using biotin labeled PNA, for better understanding of the formation and changes of glycoconjugates during oogenesis. The ultrastructure of vitellogenesis also was observed by electron microscopy for detection of the origin and track of vitelline material. In the ovary, PNA receptors appeared in the oocyte cytoplasm of the second phases of oogenesis; positive granules gradually increased from the third phase to the fourth, and they exhibited a maximum expression before the vitellogennic stage in the cytoplasm of the oocyte. From the vitellogennic to chorionation stage, positive granules gradually declined. Binding sites on follicle cells were changed with their morphological variation in every stage of oogenesis. The vitelline of A. cinerea formed within the oocyte by degrees. The results suggest that PNA receptors and yolk materials are synthesized by the oocytc at an early period. With the development of the oocyte, some exogeous materials from two sources act as PNA receptors and others take part in vitelline synthesis. One is blood lymph that offers some useful materials to the oocyte directly through follicle cell gaps; the other are follicle cells that produce and transmit some materials to oocyte to support vitellogenesis. In addition, PNA receptors secreted by follicle cells participate in the formation of yolk membrane [ Acta Zoologica Sinica 5 l （5） ： 932 - 939, 2005 ].     

Oogenesis: From Oogonia to Ovulation in the Flagfish,Jordanella floridae Goode and Bean, 1879 (Teleostei: Cyprinodontidae)

We provide histological details of the development of oocytes in the cyprinodontid flagfish, Jordanella floridae. There are six stages of oogenesis: Oogonial proliferation, chromatin nucleolus, primary growth (previtellogenesis [PG]), secondary growth (vitellogenesis), oocyte maturation and ovulation. The ovarian lamellae are lined by a germinal epithelium composed of epithelial cells and scattered oogonia. During primary growth, the development of cortical alveoli and oil droplets, are initiated simultaneously. During secondary growth, yolk globules coalesce into a fluid mass. The full‐grown oocyte contains a large globule of fluid yolk. The germinal vesicle is at the animal pole, and the cortical alveoli and oil droplets are located at the periphery. The disposition of oil droplets at the vegetal pole of the germinal vesicle during late secondary growth stage is a unique characteristic. The follicular cell layer is composed initially of a single layer of squamous cells during early PG which become columnar during early vitellogenesis. During primary and secondary growth stages, filaments develop among the follicular cells and also around the micropyle. The filaments are seen extending from the zona pellucida after ovulation. During ovulation, a space is evident between the oocyte and the zona pellucida. Asynchronous spawning activity is confirmed by the observation that, after ovulation, the ovarian lamellae contain follicles in both primary and secondary growth stages; in contrast, when the seasonal activity of oogenesis and spawning ends, after ovulation, the ovarian lamellae contain only follicles in the primary growth stage. J. Morphol. 277:1339–1354, 2016. © 2016 Wiley Periodicals, Inc.     

Oogenesis in the viviparous matrotrophic lizard Mabuya brachypoda

Oogenesis in the lizard Mabuya brachypoda is seasonal, with oogenesis initiated during May-June and ovulation occurring during July-August. This species ovulates an egg that is microlecithal, having very small yolk stores. The preovulatory oocyte attains a maximum diameter of 0.9-1.3 mm. Two elongated germinal beds, formed by germinal epithelia containing oogonia, early oocytes, and somatic cells, are found on the dorsal surface of each ovary. Although microlecithal eggs are ovulated in this species, oogenesis is characterized by both previtellogenic and vitellogenic stages. During early previtellogenesis, the nucleus of the oocyte contains lampbrush chromosomes, whereas the ooplasm stains lightly with a perinuclear yolk nucleus. During late previtellogenesis the ooplasm displays basophilic staining with fine granular material composed of irregularly distributed bundles of thin fibers. A well-defined zona pellucida is also observed. The granulosa, initially composed of a single layer of squamous cells during early previtellogenesis, becomes multilayered and polymorphic. As with other squamate reptiles, the granulosa at this stage is formed by three cell types: small, intermediate, and large or pyriform cells. As vitellogenesis progresses the oocyte displays abundant vacuoles and small, but scarce, yolk platelets at the periphery of the oocyte. The zona pellucida attains its maximum thickness during late oogenesis, a period when the granulosa is again reduced to a single layer of squamous cells. The vitellogenic process observed in M. brachypoda corresponds with the earliest vitellogenic stages seen in other viviparous lizard species with larger oocytes. The various species of the genus Mabuya provided us with important models to understand a major transition in the evolution of viviparity, the development of a microlecithal egg.     

Histological and histochemical study of female germ cell development in the dusky grouper Epinephelus marginatus (Lowe, 1834)

The developmental stages of female germ cells were analysed in a wild population of the protogynous teleost Epinephelus marginatus (Lowe, 1834). 321 wild dusky grouper females were collected in the South Mediterranean Sea during the spawning season and their ovaries analysed using histological and histochemical techniques. Oocyte morphology, nucleus-cytoplasm ratio (N/C) range, location and movements of cytoplasmic inclusions during primary growth, vitellogenesis and final oocyte maturation were described. The distribution of proteins, lipids and carbohydrates through oocyte development was also investigated in 50 females. Lipid vesicles appeared firstly in the mid ooplasm of oocytes larger than 130 microm, at the beginning of the secondary growth phase. Immediately afterwards, small carbohydrate granules (PAS and Alcian blue positive) appeared before the occurrence of the first yolk granules. Tyrosine-enriched proteins were especially evidenced in the zona radiata interna of late vitellogenic oocytes. Specific lectin binding patterns reflected characteristic differences in the content and distribution of specific sugar moieties expressed in the oocytes during vitellogenesis and final maturation. At the end of vitellogenesis and during final maturation, follicular cells, zona radiata, and cortical alveoli were characterised by a strong increase of specific binding for WGA.     

Oocyte growth in the sheepshead minnow: Uptake of exogenous proteins by vitellogenic oocytes

The structure of the vitellogenic follicle of the sheepshead minnow, Cyprinodon variegatus, is described. Follicles enlarge primarily by protein yolk accumulation (vitellogenesis) and subsequently increase in size by hydration. This study uses the electron-dense tracer, horseradish peroxidase, and a larger heterologous protein,Xenopus laevis [³H]vitellogenin, to follow the fate of exogenous proteins from the maternal circulation to yolk spheres of the growing oocyte. Materials appear to leave the perifollicular capillaries via an interendothelial route, traverse the theca and the patent intercellular channels of the follicular epithelium and the pore canals of the vitelline envelope. At the oocyte surface they are incorporated via micropinocytosis and translocated to growing yolk spheres in the peripheral ooplasm. In contrast to other studies on oocyte growth in teleosts which suggest that yolk is an autosynthetic product, this study substantiates the importance of heterosynthetic processes during oocyte growth in C. Variegatus.