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。     

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     

凡纳滨对虾卵母细胞卵黄发生的超微结构

利用电镜研究凡纳滨对虾卵母细胞卵黄发生的全过程。结果表明 :凡纳滨对虾卵黄的发生是双源性的。卵黄发生早、中期是内源性卵黄大量合成的阶段 ,卵黄发生中、后期则以外源性卵黄的合成为主。内源性卵黄主要由内质网、线粒体、核糖体、溶酶体、高尔基器等多种胞器活跃参与形成。其中数量众多的囊泡状粗面内质网是形成内源性卵黄粒的最主要的细胞器 ;部分线粒体参与卵黄粒的合成并自身最终演变为卵黄粒 ;丰富的游离核糖体合成了大量致密的蛋白质颗粒并在卵质中直接聚集融合成无膜的卵黄粒 ;溶酶体通过吞噬、消化内含物来形成卵黄粒和脂滴 ,且方式多样 ;高尔基器不直接参与形成卵黄粒。外源性卵黄主要通过卵质膜的微吞饮活动从卵周隙或卵泡细胞中摄取外源物质来形成     

锯缘青蟹卵黄发生期卵母细胞和卵泡细胞之间的结构变化

通过电镜研究了锯缘青蟹二次卵巢发育过程中卵黄发生期（分为初期和后期）卵母细胞表面的结构和胞质的变化。卵黄发生初期分为：内源性卵黄发生阶段和有卵泡细胞直接参与的外源性卵黄合成阶段,前者特征为：在卵母细胞中充满了内质网泡,在泡内有不同程度的卵黄物质合成,此时在卵母细胞的表面区域,可见很多卵泡细胞向卵母细胞表面迁移,并包围卵母细胞。后者其特征是在卵母细胞的表面,有大量的胞饮小泡出现在卵膜的内面,随着两细胞表面膜的逐步融合和胞饮作用加强最后形成链锁状结构,胞质中靠近卵质周围有卵黄体的积极合成和大更换 脂肪滴积累,在此阶段的后期,卵泡细胞质已基本吸收完毕,卵泡细胞膜和卵母细胞膜融合,某些界面已无膜结构。卵黄发生后期在亲蟹孵出幼体后的第11d至第27d基本结束,此期也主要以外源性卵黄发生为主,在卵母细胞的周围,卵泡细胞迅速扩大,其间分布着大量的大小不同的囊泡和线粒体,在接近卵母细胞表面,还常可见大量的脂肪滴存在。卵泡细胞与卵母细胞间其膜结构完全消失,从而可使滤泡大片细胞质直接融入卵母细胞中,以后随着卵黄发生的进一步发展,卵母细胞与卵泡细胞的交界面逐步形成一个网状的膜结构屏障,同时在卵巢中可见正在降解的卵母细胞,在卵黄发生近结束以后,在卵母细胞的表面,逐步形成两层卵膜,这时的卵母细胞质中几乎充满了卵黄体和脂肪滴。     

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。     

北京油葫芦卵黄物质形成的超微结构观察

以蟋蟀科的北京油葫芦Teleogryllusmitratrus(Burmeister)为材料 ,对其卵子发生的卵黄物质形成过程的超微结构进行了观察。根据电镜观察结果分析 ,北京油葫芦卵黄构成有卵母细胞内部物质与外部物质参与。卵黄发生初期 ,主要以卵母细胞自身合成为主 ,随着卵母细胞发育的进行 ,有外源物质介入卵黄合成之中。它包括两部分物质来源 :一部分是由血淋巴通过滤泡细胞间隙向卵母细胞提供合成卵黄物质 ;另一部分则由滤泡细胞通过指状微绒毛以多泡小体和多片小体的形式向卵母细胞提供合成卵黄的物质。     

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。     

罗氏沼虾卵母细胞细胞器与卵黄发生的关系

作者研究了罗氏沼虾卵母细胞细胞器与卵黄发生的关系。细胞核物质通过核孔与细胞质进行物质交换,由核仁而来的核糖体前体物在细胞质中形成大量的核糖体。一部分核糖体附着在内质网囊泡的膜上,形成粗糙型内质网。粗糙型内质同囊泡具有合成和运输卵黄物质的功能。一部分核糖体是游离的,它们可在细胞质中合成致密蛋白质小颗粒,随后这些颗粒逐渐聚集融合成致密的卵黄粒。溶酶体可吞噬这些由游离核糖体形成的蛋白质颗粒团或其它膜状物,并将它们消化、融合,从而形成大的卵黄粒。部分线粒体参与了卵黄粒的合成并最终演变成卵黄体。微丝也参与了卵黄粒的形成。       

南美白对虾卵子发生的组织学

采用组织学方法研究了南美白对虾的卵子发生过程,根据卵细胞大小、核仁形态、卵黄粒的有无、皮质棒的出现以及卵母细胞与滤泡细胞的关系,将南美白对虾的卵子发生划分为卵原细胞、卵黄发生前的卵母细胞和卵黄发生的卵母细胞三个时期,并描述了各期卵细胞的形态特征。     

剑尾鱼卵子发生的组织学观察

应用光学显微镜对卵胎生硬骨鱼类剑尾鱼(Xiphophorus helleri)卵巢的组织结构进行了观察。结果显示,剑尾鱼卵子的发育过程可划分为6个时相。Ⅰ时相的卵母细胞呈原始分化状态,细胞外具一层细胞质膜。Ⅱ时相卵母细胞外不仅具有质膜,而且还包绕一层滤泡细胞。Ⅲ时相和Ⅳ时相的卵母细胞分化明显,胞质内开始积累脂滴和卵黄颗粒。Ⅴ时相为成熟卵子,卵子的卵膜极薄,胞质内含有丰富的脂滴和卵黄。Ⅵ时相卵母细胞进入退化期,滤泡细胞从卵周向中央突入,卵黄被完全吸收,滤泡细胞自身也变得肥大。结果表明,剑尾鱼卵巢中卵母细胞的发育是不同步的。     

Ultrastructure of the ovarian follicles in the placentotrophic Andean lizard of the genus Mabuya (Squamata: Scincidae)

We studied the ultrastructural organization of the ovarian follicles in a placentotrophic Andean lizard of the genus Mabuya. The oocyte of the primary follicle is surrounded by a single layer of follicle cells. During the previtellogenic stages, these cells become stratified and differentiated in three cell types: small, intermediate, and large globoid, non pyriform cells. Fluid‐filled spaces arise among follicular cells in late previtellogenic follicles and provide evidence of cell lysis. In vitellogenic follicles, the follicular cells constitute a monolayered granulosa with large lacunar spaces; the content of their cytoplasm is released to the perivitelline space where the zona pellucida is formed. The oolemma of younger oocytes presents incipient short projections; as the oocyte grows, these projections become organized in a microvillar surface. During vitellogenesis, cannaliculi develop from the base of the microvilli and internalize materials by endocytosis. In the juxtanuclear ooplasm of early previtellogenic follicles, the Balbiani's vitelline body is found as an aggregate of organelles and lipid droplets; this complex of organelles disperses in the ooplasm during oocyte growth. In late previtellogenesis, membranous organelles are especially abundant in the peripheral ooplasm, whereas abundant vesicles and granular material occur in the medullar ooplasm. The ooplasm of vitellogenic follicles shows a peripheral band constituted by abundant membranous organelles and numerous vesicular bodies, some of them with a small lipoprotein core. No organized yolk platelets, like in lecithotrophic reptiles, were observed. Toward the medullary ooplasm, electron‐lucent vesicles become larger in size containing remains of cytoplasmic material in dissolution. The results of this study demonstrate structural similarities between the follicles of this species and other Squamata; however, the ooplasm of the mature oocyte of Mabuya is morphologically similar to the ooplasm of mature oocytes of marsupials, suggesting an interesting evolutionary convergence related to the evolution of placentotrophy and of microlecithal eggs. J. Morphol., 2010. © 2010 Wiley‐Liss, Inc.     

The ultrastructure of oogenesis and yolk formation in Labidocera aestiva (Copepoda: Calanoida)

Yolk formation in the oocytes of the free-living, marine copepod, Labidocera aestiva (order Calanoida) involves both autosynthetic and heterosynthetic processes. Three morphologically distinct forms of endogenous yolk are produced in the early vitellogenic stages. Type 1 yolk spheres are formed by the accumulation and fusion of dense granules within vesicular and lamellar cisternae of endoplasmic reticulum. A granular form of type 1 yolk, in which the dense granules within the cisternae of endoplasmic reticulum do not fuse, appears to be synthesized by the combined activity of endoplasmic reticulum and Golgi complexes. Type 2 yolk bodies subsequently appear in the ooplasm but their formation could not be attributed to any particular oocytic organelle. In the advanced stages of vitellogenesis, a single narrow layer of follicle cells becomes more developed and forms extensive interdigitations with the oocytes. Extra-oocytic yolk precursors appear to pass from the hemolymph into the follicle cells and subsequently into the oocytes via micropinocytosis. Pinocytotic vesicles fuse in the cortical ooplasm to form heterosynthetically derived type 3 yolk bodies.     

Ultrastructural study of oogenesis in the acoel turbellarian Convoluta

An ultrastructural investigation of oogenesis has been carried out on the acoel turbellarian Convoluta psammophyla. Developing female germ cells are not contained in well delimited ovaries but are freely distributed in the parenchyma and are surrounded by narrow cytoplasmic projections of accessory-follicle cells. Oogenesis can be divided into two periods, the previtellogenic and the vitellogenic phase. In the first period the oocyte undergoes a number of cell differentiations necessary for the intense biosynthetic activity of the second period. The ample development of nucleolus, ribosomes, endoplasmic reticulum and Golgi complexes along with the appearance of large lipid droplets and clusters of electron dense granules characterize the previtellogenic phase. The formation of yolk globules is the main feature of the second period of oogenesis. It occurs by an autosynthetic mechanism involving endoplasmic reticulum and Golgi complexes, since no endocytotic activity has been detected in the developing oocyte. The electron dense granules apparently move towards the cortical ooplasm during the late vitellogenic phase and take part in egg covering formation. Hypotheses on the role of follicle cells and on the phylogenetic significance of a comparative analysis of egg inclusions with homologous structures of other Turbellaria are suggested.     

Fine structure of the developing oocytes in adultArgas (Persicargas) arboreus (Ixodoidea: Argasidae)

The structure of the developing oocytes in the ovary of unfed and fed femaleArgas (Persicargas) arboreus is described as seen by scanning (SEM) and transmission (TEM) electron microscopy. The unfed female ovary contains small oocytes protruding onto the surface and its epithelium consists of interstitial cells, oogonia and young oocytes. Feeding initiates oocyte growth through the previtellogenic and vitellogenic phases of development. These phases can be observed by SEM in the same ovary.The surface of isolated, growing oocytes is covered by microvilli which closely contact the basal lamina investing the ovarian epithelium and contains a shallow, circular area with cytoplasmic projections and a deep pit, or micropyle, at the epithelium side. In more advanced oocytes the shell is deposited between microvilli and later completely covers the surface.Transmission EM of growing oocytes in the previtellogenic phase reveals nuclear and nucleolar activity in the emission of dense granules passing into the cytoplasm and the formation of surface microvilli. The cell cytoplasm is rich in free ribosomes and polysomes and contains several dictyosomes associated with dense vesicles and mitochondria which undergo morphogenic changes as growth proceeds. Membrane-limited multivesiculate bodies, probably originating from modified mitochondria, dictyosomes and ribosomal aggregates, are also observed. Rough endoplasmic reticulum is in the form of annulate lamellae. During vitellogenesis, proteinaceous yolk bodies are formed by both endogenous and exogenous sources. The former is involved in the formation of multivesicular bodies which become primary yolk bodies, whereas the latter process involves internalization from the haemolymph through micropinocytosis in pits, vesicles and reservoirs. These fuse with the primary yolk bodies forming large yolk spheres. Glycogen and lipid inclusions are found in the cytoplasm between the yolk spheres.     

Immunolocalization of estrogen receptor α in Neomysis japonica oocytes and follicle cells during ovarian development

Estrogen induces oocytes development and vitellogenesis in crustacean by interacting with estrogen receptor (ER) subtypes. In the present study, we detect for the first time the ERα in oocytes and follicle cells and hepatopancreas cells of mysis by immunohistochemistry using a specific ERα antibody. ERα was mainly localized in the nuclei of oocytes and follicle cells, while mainly detected in nuclei of oogonia (OG), previtellogenic oocyte (PR) and endogenous vitellogenic oocyte (EN) at previtellogenic and early vitellogenic stage (I-early III). Follicle cells in all stages of ovary (all vitellogenic stages) showed strong ERα positive reaction, and they were able to gradually move to oocytes during the development of oocytes. In addition, ERα was also localized in the nuclei and cytoplasm of four hepatopancreas cells (including E-, R-, F- and B-cell) in all ovary stages. These findings suggest, for the first time to our knowledge, that there could be a close link between oogenesis, follicle cells, hepatopancreas cells and endocrine regulation, and estrogens might be involved in the regulation of oocytes at early ovarian stage in mysis.     

Degeneration of germ line cells in amphibian ovary

Ogielska, M., Rozenblut, B., Augustyńska, R., Kotusz, A. 2010. Degeneration of germ line cells in amphibian ovary. —Acta Zoologica (Stockholm) 91 : 319–327 We studied the morphology of degenerating ovarian follicles in juvenile and adult frogs Rana temporaria, Rana lessonae and Rana ridibunda. Degeneration of primordial germ cells was never observed and was extremely rare in oogonia and early oocytes in a cyst phase in juveniles. Previtellogenic oocytes were rarely affected. Three main types of atresia were identified. In type I (subdivided into stages A–D), vitellogenic oocytes are digested by proliferating follicle cells that hypertrophy and become phagocytic. A – germinal vesicle shrinks, nucleoli fuse, oocyte envelope interrupts, and follicular cells hypertrophy; B – follicular cells multiply and invade the oocyte; C – entire vesicle is filled by phagocytic cells; D – degenerating phagocytes accumulate black pigment. Type II is rare and resembles breakdown of follicles and release of ooplasm. In type III, observed in previtellogenic and early vitellogenic oocytes, ooplasm and germinal vesicle shrink, follicle cells do not invade the vesicle, and condensed ooplasm becomes fragmented. The residual oogonia in adult ovaries (germ patches) multiply, but soon degenerate.     

Ovarian maturation and oogenesis in the blue swimmer crab,Portunus pelagicus (Decapoda: Portunidae)

The study was aimed at understanding the process of reproduction and the changes happening in the ovary of Portunus pelagicus during maturation, which would be useful for its broodstock development for hatchery purposes. For that, tissue samples from different regions of the ovary at various stages of maturation were subjected to light and electron microscopy, and based on the changes revealed and the differences in ovarian morphology, the ovary was divided into five stages such as immature (previtellogenic oocytes), early maturing (early vitellogenic oocytes), late maturing (late vitellogenic oocytes), mature (vitellogenic oocytes), and spent (resorbing oocytes). The ovarian wall comprised of an outermost thin pavement epithelium, a middle layer of connective tissue, and an innermost layer of germinal epithelium. The oocytes matured as they moved from the centrally placed germinal zone toward the ovarian wall. The peripheral arrangement of nucleolar materials and the high incidence of cell organelles during the initial stages indicated vitellogenesis I. Movement of follicle cells toward oocytes in the early maturing stage and low incidence of mitochondria and endoplasmic reticulum in the ooplasm during late vitellogenic stage marked the commencement and end of vitellogenesis II, respectively. Yolk granules at various stages of development were seen in the ooplasm from late vitellogenic stage onwards. The spent ovary had an area with resorbing oocytes and empty follicle cells denoting the end of one reproductive cycle and another area with oogonial cells and previtellogenic oocytes indicating the beginning of the next.     

Ultrastructural observations of previtellogenic ovarian follicles of the caecilians Ichthyophis tricolor and Gegeneophis ramaswamii

The ultrastructural organization of the previtellogenic follicles of the caecilians Ichthyophis tricolor and Gegeneophis ramaswamii, of the Western Ghats of India, were observed. Both species follow a similar seasonal reproductive pattern. The ovaries contain primordial follicles throughout the year with previtellogenic, vitellogenic, or postvitellogenic follicles, depending upon the reproductive status. The just-recruited primordial follicle includes an oocyte surrounded by a single layer of follicle and thecal cells. The differentiation of the theca into externa and interna layers, the follicle cells into dark and light variants, and the appearance of primordial yolk platelets and mitochondrial clouds in the ooplasm mark the transition of the primordial follicle into a previtellogenic follicle. During further development of the previtellogenic follicle the following changes occur: i) the theca loses distinction as externa and interna; ii) all the follicle cells become the dark variant and increase in the complexity of ultrastructural organization; iii) the nucleus of the oocyte transforms into the germinal vesicle and there is amplification of the nucleoli; iv) the primordial yolk platelets of the cortical cytoplasm of the oocyte increase in abundance; v) the mitochondrial clouds fragment and the mitochondria move away from the clouds, leaving behind the cementing matrix, which contains membrane-bound vesicles of various sizes, either empty or filled with a lipid material; vi) the perivitelline space appears first as troughs at the junctional points between the follicle cells and oocyte, which subsequently spread all around to become continuous; vii) macrovilli and microvilli develop from the follicle cells and oocyte, respectively; and viii) the precursor material of the vitelline envelop arrives at the perivitelline space. The sequential changes in the previtellogenic follicles of two species of caecilians are described.     

Formation of an egg envelope in the pycnogonid, Propallene longiceps (Pycnogonida, Callipallenidae)

Vitellogenic oocytes of all pycnogonids studied so far contain dilated elements of the endoplasmic reticulum, filled with characteristic electron-dense bodies. During vitellogenesis these bodies fuse and form larger, almost spherical granules that were traditionally interpreted as nascent yolk granules. Here, we present the results of ultrastructural investigations of previtellogenic and early vitellogenic oocytes of Propallene longiceps (Pycnogonida, Callipallenidae). We show that the intra-cisternal bodies/granules of pycnogonids are not involved in vitellogenesis but contain macromolecules that are released from the oocyte and contribute to the formation of an egg envelope. The obtained results are discussed in a phylogenetic context. We suggest that the presence of the intra-cisternal electron-dense bodies in the oocyte cytoplasm represents a plesiomorphic character of arthropods inherited from the arthropod ancestor.