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     

荞麦子叶发育过程中聚合的粗糙内质网

在荞麦（Fagopyrum esculentum Moench.）子叶的发育过程中观察到粗糙内质网的聚合现象,是一些粗糙内质网平行或成环形排列形成的。这些聚合的粗糙内质网附近通常有蛋白质团块存在,其形状不规则,无膜包被,与周围细胞质的界限不清。推测这些蛋白质团块不是在液泡或膨大的粗糙内质网囊泡中积累形成的,而是由聚合的粗糙内质网直接将合成的蛋白质分泌到细胞质中形成的。这种形式提高了子叶中贮存蛋白质的积累效率。     

降香黄檀次生韧皮部薄壁组织细胞液泡蛋白质的形成和积累的超微结构研究

热带豆科树木降香黄檀（Ｄａｌｂｅｒｇｉａｏｄｏｒｉｆｅｒａ）落叶时期末端小枝次生韧皮部薄壁组织细胞的中央液泡中有大量的贮藏蛋白质。用透射电镜技术研究了这些蛋白质的形成和积累。在这些细胞的周缘细胞质中有丰富的游离核糖体和许多粗糙内质网（ＲＥＲ）。在这些核糖体之间,总是分散着纤维状物质,有时还出现一些类似液泡蛋白质的结构。常常看到与ＲＥＲ相连的含有稀疏纤维状物质的小泡,以及含有类似液泡蛋白质的形状不规则的小泡和含有更多类似液泡蛋白质的光滑小泡。从连接ＲＥＲ的小泡到形状不规则小泡,再到光滑小泡,可能存在个体发生的连续性。通过内吞作用细胞质中的纤维状物质和核糖体以及其它结构被并入上述３种小泡。光滑小泡通常与中央液泡的膜融合而释放其内含物到液泡中。     

东方扁虾卵子发生的超微结构

根据卵细胞的形态、内部结构特征及卵母细胞与滤泡细胞之间的关系,东方扁虾的卵子发生可划分为卵原细胞、卵黄发生前卵母细胞、卵黄发生卵母细胞和成熟卵母细胞等四个时期。卵原细胞胞质稀少,胞器以滑面内质网为主。卵黄发生前卵母细胞核明显膨大,特称为生发泡;在靠近核外膜的胞质中可观察到核仁外排物。卵黄发生卵母细胞逐渐为滤泡细胞所包围;卵黄合成旺盛,胞质中因而形成并积累了越来越多的卵黄粒。东方扁虾卵母细胞的卵黄发生是二源的。游离型核糖体率先参与内源性卵黄合成形成无膜卵黄粒。粗面内质网是内源性卵黄形成的主要胞器。滑面内质网、线粒体和溶酶体以多种方式活跃地参与卵黄粒形成。卵周隙内的外源性物质有两个来源:滤泡细胞的合成产物和血淋巴携带、转运的卵黄蛋白前体物。这些外源性物质主要通过质膜的微吞饮作用和微绒毛的吸收作用这两种方式进入卵母细胞,进而形成外源性卵黄。内源性和外源性的卵黄物质共同参与成熟卵母细胞中富含髓样小体的卵黄粒的形成。卵壳的形成和微绒毛的回缩被认为是东方扁虾卵母细胞成熟的形态学标志。       

西瓜种子发育和萌发过程中子叶细胞超微结构的变化

西瓜种子子叶内贮存物质开始积累时,细胞质内有大量核糖体、质体、线粒体,内质网片段和囊泡,种子脱水期至成熟期,细胞器的数量减少,成熟种子子叶细胞的细胞壁不连续,几乎观察不到细胞器的存在,种子萌发过程中内质网,线粒体,质体的数目逐渐增多,叶肉细胞的质体发育成叶绿体,种子形成过程中,在子叶细胞大液泡分隔的同时,膨胀的内质网囊泡内积累蛋白质（直径0.1-0.4μm),这些小的蛋白质球体最终进入液泡形成大的蛋白体（直径1-3μm);萌发种子贮存蛋白质被水解的同时,一些脂体进入液泡并被分解,同时液泡融合;脂类物质开始积累的时间早于蛋白质,积累的量较蛋白质多,但在萌发种子中被彻底水解的时间晚于蛋白质,淀粉粒的数量在种子形成时减少,种子萌发时在表皮细胞和叶肉细胞内都重新合成。     

降香黄檀次生韧皮部薄壁组织细胞液泡蛋白质的形成和积累的超…

热带豆科树木降香黄檀落叶时期末端小枝次生韧皮部薄壁组织细胞的中央液泡中有大量的贮藏蛋白质。用透射电镜技术研究了这些蛋白质的形成和积累。在这些细胞的周缘细胞质中有的游离核糖体例和许多粗糙内质网（ＲＥＲ）。在这些 核糖体之间,总是分散着纤维状物质,有时还出现一些类似液泡蛋白质的结构。     

高等植物花色苷的液泡摄取机制

综述了高等植物细胞中花色苷被液泡摄取的机制。花色苷通过细胞质中定位于粗糙内质网细胞质面的多酶复合体合成后被膜包裹形成囊泡。这些囊泡主要向液泡移动,在移动中相互融合形成更大囊泡,最终将花色苷带到液泡膜的表面。在大多数情况下,花色苷经过液泡膜上的各种载体被迅速运进液泡。另外两种较少的是:(1)囊泡直接与液泡融合;(2)液泡膜自主形成大的管状内陷,使囊泡在内陷处指向液泡内腔"发芽"。在上述种种可能的具体过程中,花色苷以非修饰或修饰两种形式被摄入液泡。花色苷跨液泡膜运送可能通过4种模型实现,即由ATP结合盒型的载体介导、由依赖pH梯度的载体介导、由24-kD液泡蛋白前体衍生的蛋白质介导和由多重药物和有毒化合物排出家族的载体介导。据推测,不同植物利用不同的摄取机制将花色苷积累在液泡中,而多重机制也可能被单个植物种同时使用。     

长江华溪蟹纳精囊超微结构的研究

利用电镜技术,对长江华溪蟹的纳精囊进行了研究。结果表明:在纳精囊上皮的顶分泌型腺细胞中,充满大量高尔基体和粗面内质网的潴泡和囊泡。泡中含有絮状或颗粒状分泌物。潴泡和囊泡先是单独存在,最后聚集在一起,形成大的分泌颗粒后排出囊腔。核糖体比比皆是。线粒体数量较大,作为一种载体参与了分泌物的形成。细胞化学显示,分泌物中含有蛋白质、脂肪和少量糖类。结论:纳精囊上皮的顶分泌型腺细胞具有积极的分泌活动。       

山珊瑚内生菌根的研究

山珊瑚内生菌根真菌在人工培养基上分离培养不能生长。侵染菌丝由山珊瑚侧根表皮侵入,在皮层中部形成侵染通道。进入皮层细胞后形成丛枝吸器。被侵染的寄主细胞仍是生活细胞,但细胞质变稀,蛋白质、RNA含量很少。近中柱的6至8层皮层细胞壁增厚并木质化。丛枝吸器含有丰富的红淀粉,少量的中性脂肪、碱性蛋白质和D NA。菌丝丛枝干幼龄期电子致密,周围有界面物质包围,在衰老时泡囊化,界面物质亦瓦解。吸器由五边形结构组成。寄主细胞在被侵染前含淀粉质体、线粒体、高尔基体、内质网和核糖体,在被侵染后,淀粉质体消失,其它细胞器数量明显减少,而主要是微丝和小液泡及多泡体。文章讨论了山珊瑚菌根与天麻和泡囊-丛枝菌根的异同。     

知母绒毡层和乌氏体细微结构的研究

从个体发育来看,知母绒毡层具有3个明显的特点;（1）在小孢子母细胞阶段,绒毡层细胞中有丰富的细胞器,如粗糙内质网,脂体,造粉体和小泡等,粗糙内质网－脂体－小泡常常联系在一起,形成细胞器复合体。（2）在单核花粉阶段,绒毡层细胞质中出现大量小泡,它们可以融合成小泡,而且在小泡或大泡中开始沉积与脂体电子密度相似的亲锇物质,这些物质或者充满整个小泡,或者沉积在小泡周缘,此刻,也是乌氏体形成并达到高峰的阶段。（3）在成熟花粉阶段,绒毯层细胞几乎被具膜束缚的小型脂粒和巨型脂体所占据,这些亲锇物质是质体起源的,也许它们是花粉鞘的先质。     

Ultrastructural studies on the formation of yolk granules in the statoblast of a fresh-water bryozoan,Pectinatella gelatinosa

The formation of protein-carbohydrate yolk in the statoblast of a fresh-water bryozoan, Pectinatella gelatinosa, was studied by electron microscopy. Two types (I and II) of yolk cells were distinguished. The type I yolk cells are mononucleate and comprise a large majority of the yolk cells. The type II yolk cells are small in number; they become multinucleate by fusion of cells at an early stage of vitellogenesis. In both types of yolk cells, electron-dense granules (dense bodies) are formed in Golgi or condensing vacuoles, which are then called yolk granules. For the formation of yolk granules, the following processes are considered: 1. Yolk protein is synthesized in the rough-surfaced endoplasmic reticulum (RER) of the yolk cells. 2. The synthesized protein condenses in the cisternal space of the RER and is packaged into small oval swellings, which are then released from the RER as small vesicles (Golgi vesicles, 300-600 A in diameter). 3. The small vesicles fuse with one another to form condensing vacuoles, or with pre-existing growing yolk granules. 4. In the matrix of the condensing vacuoles or growing yolk granules, electron-dense fibers are fabricated and then arranged in a paracrystalline pattern to form the dense body. 5. After the dense body reaches its full size, excess membrane is removed and eventually the yolk granules come to mature. Toward the end of vitellogenesis of the yolk cells, the cytoplasmic organelles are ingested by autophagosomes derived from multivesicular bodies and disappear.     

Synthesis,secretion and immunoelectron microscopic demonstration of apolipoprotein B-containing lipoprotein particles in the visceral rat yolk sac

Summary Electron microscopic investigations on the involvement of the fetal membranes of the rat (visceral yolk sac) in the lipid metabolism revealed the occurrence of lipoprotein-sized particles located in cisternal Golgi stacks, Golgi vesicles and secretory vesicles of the cells of the visceral yolk sac epithelium as well as in distended areas of the intercellular space between adjacent epithelial cells. Application of the protein A-gold technique with specific anti-apoB antiserum resulted in a specific location of immunogold both over the different compartments of the lipoprotein pathway (RER, Golgi complex, secretory vesicles) as well as over the distended intercellular spaces, thus confirming these particles to be lipoproteins in nature. Isolated visceral epithelial cells prepared by a tryptic digestion method exhibited some ultrastructural alterations, such as a loss of apical brush border, a change from columnar to spherical cell shape, a decrease in phagolysosomes, but an increase in autophagosomal structures after 6 h incubation at a vitality rate of at least 85%. Within this period the epithelial cells secreted measurable amounts of apoB-containing lipoproteins into the medium floating in the density classes d<1.006 g/ml, d=1.006–1.020 g/ml and d=1.020–1.064 g/ml. The production of the lipoproteins was partly inhibited by cycloheximide indicating the secretion of particles with preformed as well as newly synthesized apoB. Negative staining of the particles revealed an average diameter of 34 nm of VLDL, 31 nm of IDL and 24 nm of LDL. In summary, our studies demonstrate that in the feto-placental unit of the rat the fetal membranes are capable of synthesizing and secreting lipoproteins. The cells of the visceral yolk sac epithelium were shown to be the producers of apoB-containing particles.Abbreviations apo apolipoprotein - ER endoplasmic reticulum - IDL intermediate density-lipoprotein - LDL low density-lipoprotein - VLDL very low density-lipoprotein - PBS phosphate-buffered salt solution - RER rough endoplasmic reticulum - TEM transmission electron microscopy     

Synthesis, secretion and immunoelectron microscopic demonstration of apolipoprotein B-containing lipoprotein particles in the visceral rat yolk sac.

Electron microscopic investigations on the involvement of the fetal membranes of the rat (visceral yolk sac) in the lipid metabolism revealed the occurrence of lipoprotein-sized particles located in cisternal Golgi stacks, Golgi vesicles and secretory vesicles of the cells of the visceral yolk sac epithelium as well as in distended areas of the intercellular space between adjacent epithelial cells. Application of the protein A-gold technique with specific anti-apoB antiserum resulted in a specific location of immunogold both over the different compartments of the lipoprotein pathway (RER, Golgi complex, secretory vesicles) as well as over the distended intercellular spaces, thus confirming these particles to be lipoproteins in nature. Isolated visceral epithelial cells prepared by a tryptic digestion method exhibited some ultrastructural alterations, such as a loss of apical brush border, a change from columnar to spherical cell shape, a decrease in phagolysosomes, but an increase in autophagosomal structures after 6 h incubation at a vitality rate of at least 85%. Within this period the epithelial cells secreted measurable amounts of apoB-containing lipoproteins into the medium floating in the density classes d less than 1.006 g/ml, d = 1.006-1.020 g/ml and d = 1.020-1.064 g/ml. The production of the lipoproteins was partly inhibited by cycloheximide indicating the secretion of particles with performed as well as newly synthesized apoB. Negative staining of the particles revealed an average diameter of 34 nm of VLDL, 31 nm of IDL and 24 nm of LDL. In summary, our studies demonstrate that in the feto-placental unit of the rat the fetal membranes are capable of synthesizing and secreting lipoproteins. The cells of the visceral yolk sac epithelium were shown to be the producers of apoB-containing particles.     

Formation of intracellular particles by hepatitis B virus large surface protein.

Hepatitis B virus small surface protein is synthesized as a transmembrane protein of the rough endoplasmic reticulum (RER) and then buds into the lumen in the form of subviral particles that are secreted. The closely related large surface protein is also targeted to the RER but is retained in a pre-Golgi compartment and cannot be secreted. It has been assumed that the large surface protein remains as a transmembrane RER protein and hence cannot form particles, possibly because of binding to a host factor on the cytosolic face of the RER membranes. We have reexamined this question and found the following results. (i) The retained large surface protein is associated not with RER but, rather, with a more distal compartment. (ii) Electron microscopy reveals intravesicular 20-nm particles, similar to those formed by the small surface protein. (iii) The large surface protein colocalizes with and binds to calnexin, an ER chaperone protein. Therefore, our results indicate that the large surface protein is capable of budding and forming particles, and hence its intracellular retention cannot be attributed to a cytosolic factor. We interpret the data as evidence that the large surface protein is retained by virtue of interacting with calnexin, a component of what is considered the quality control mechanism of the ER.     

Receptor activity of rotavirus nonstructural glycoprotein NS28.

Rotavirus morphogenesis involves the budding of subviral particles through the rough endoplasmic reticulum (RER) membrane of infected cells. During this process, particles acquire the outer capsid proteins and a transient envelope. Previous immunocytochemical and biochemical studies have suggested that a rotavirus nonstructural glycoprotein, NS28, encoded by genome segment 10, is a transmembrane RER protein and that about 10,000 Mr of its carboxy terminus is exposed on the cytoplasmic side of the RER. We have used in vitro binding experiments to examine whether NS28 serves as a receptor that binds subviral particles and mediates the budding process. Specific binding was observed between purified simian rotavirus SA11 single-shelled particles and RER membranes from SA11-infected monkey kidney cells and from SA11 gene 10 baculovirus recombinant-infected insect cells. Membranes from insect cells synthesizing VP1, VP4, NS53, VP6, VP7, or NS26 did not possess binding activity. Comparison of the binding of single-shelled particles to microsomes from infected monkey kidney cells and from insect cells indicated that a membrane-associated component(s) from SA11-infected monkey kidney cells interfered with binding. Direct evidence showing the interaction of NS28 and its nonglycosylated 20,000-Mr precursor expressed in rabbit reticulocyte lysates and single-shelled particles was obtained by cosedimentation of preformed receptor-ligand complexes through sucrose gradients. The domain on NS28 responsible for binding also was characterized. Reduced binding of single-shelled particles to membranes was seen with membranes treated with (i) a monoclonal antibody previously shown to interact with the C terminus of NS28, (ii) proteases known to cleave the C terminus of NS28, and (iii) the Enzymobead reagent. VP6 on single-shelled particles was suggested to interact with NS28 because (i) a monoclonal antibody to the subgroup I epitope on VP6 reduced particle binding, (ii) a purified polyclonal antiserum raised against recombinant baculovirus-produced VP6 reduced ligand binding, and (iii) a monoclonal antibody to a conserved epitope on VP6 augmented ligand binding. These experimental data provide support for the hypothesized receptor role of NS28 before the budding stage of rotavirus morphogenesis.     

Ultrastructural changes in maturing pollen grains ofApium nodiflorum L. (Apiaceae), with special reference to the endoplasmic reticulum

Summary The ultrastructural events in 3-cellular pollen grains ofApium nodiflorum L. are investigated during pollen maturation. Three distinct developmental stages are distinguished from the formation of sperm cells up to anthesis, whereby the rough endoplasmic reticulum (RER) is mainly involved. The most conspicious form is the highly dilated RER in the vegetative cytoplasm of the youngest pollen grains, which changes to vesicular RER in the following stage. In mature pollen grains the RER has a narrow cisternal configuration and often forms stacks. Pollen activation is preceded by the accumulation of polysaccharide particles.     

Yolk organelles and their membranes during vitellogenesis ofXenopus oocytes

Summary The yolk platelets ofXenopus laevis have been studied by thin-section and freeze-fracture electron microscopy to characterize the boundary membrane during yolk formation. Throughout vitellogenesis, large yolk platelets are in close contact with smaller nascent yolk organelles. Two types of primordial yolk platelets (I and II) have been discriminated. After membrane fusion these precursors can be completely incorporated into the main body of existing platelets, numerous yolk crystals then merge and form one uniformly stratified core. Lipid droplets are tightly attached to the membrane at all developmental stages of yolk platelets. A direct connection of endoplasmic reticulum to the membranes of yolk platelets was not observed. On freezeetching replicas, yolk-platelet membranes present fracture faces with intramembranous particles (IMP) of various sizes and a heterogeneous distribution of approximately 200–600 IMP/μm² at the E face, and 1200–2100 IMP/μm² at the P face. Again, this presentation of the membrane exhibits neither anastomoses to the endoplasmic reticulum, nor caveolae that exclude the uptake of yolk-containing vesicles into these yolk organelles. Proteinaceous yolk platelets tend to fracture along their periphery through the superficial layers.     

Biosynthesis of high density lipoprotein by chicken liver: nature of nascent intracellular high density lipoprotein

Young chickens were administered L-[(3)H]leucine and after 10 or 30 min the livers were removed and fractioned into rough (RER) and smooth (SER) endoplasmic reticulum fractions and into light, intermediate, and heavy golgo cell fractions. The labeled high density lipoprotein (HDL), contained within these intracellular organelles was isolated either by immunoprecipitation using rabbit antiserum to rooster HDL, or by ultracentrifugal glotation between densities 1.063 and 1.21 g/ml. The radioactive apoproteins of nascent HDL were analyzed by SDS PAGE and detected by fluorography. Analyses of radioactive apoproteins obtained by immunoprecipitation from the contents of the RER, the SER, and the three golgi complex fractions revealed only one apoprotein, A1. The C peptide present in serum HDL was not detected intracellularly. The radioactive apoprotein A1 which is present within the cisternae of the RER and the SER fractions failed to float, whereas apoprotein A1, present within the golgi apparatus, readily floated between densities 1.063 and 1.21 g/ml. The HDL particles, isolated by flotation from the golgi apparatus content, were further characterized by lipid and protein analyses and by electron microscopy. Golgi HDL particles have the same density as serum HDL. On a percentage basis, golgi HDL contains less protein and more phospholipids than does serum HDL. Morphologically, golgi HDL is different in appearance from serum HDL. It is more heterogeneous in size, with most of the particles ranging 8.3-25 nm in diameter. The spherical particles contain small membrane tails. Occasionally, a few disk-shaped bilayer structures are also found within the golgi apparatus. These studies show that the newly synthesized apoprotein A1, present within the RER and the SER cell fractions, is not fully complexed with lipid and that apoprotein A1 does not acquire sufficient lipid to float at the proper HDL density until it enters the golgi apparatus. The difference in chemical composition and the heterogeneous size of golgi HDL may be attributed to the different stages of HDL maturation.     

Biosynthesis of lipoprotein: location of nascent apoAI and apoB in the rough endoplasmic reticulum of chicken hepatocytes

Our previous studies showed that in hepatic RER of young chickens, nascent apoAI is not associated with lipoprotein particles and only becomes part of these lipoprotein structures in the Golgi. In this study, we have used three different methodologies to determine the locations of apoAI and apoB in the RER and compared them to that of albumin. Immunoelectron microscopic examination of the RER cell fractions showed that both apoAI and apoB were associated only with the RER membrane whereas albumin was located both within the lumen and on the limiting membrane of the vesicles. To examine the possibility of membrane integration of nascent apoAI and apoB in the RER, we administered L-[3H]leucine to young chickens for 10 min, isolated RER, treated this cell fraction with buffers of varying pH, and measured the release of radioactive albumin, apoAI, and apoB. The majority of nascent apoAI (64%), nascent apoB (100%), and nascent albumin (97%) was released from RER vesicles at pH 11.2, suggesting that, like albumin, apolipoproteins are not integrated within the membrane. To determine if nascent apoproteins are exposed to the cytoplasmic surface, we administered L-[3H]leucine to young chickens and at various times isolated RER and Golgi cell fractions. Radioactive RER and Golgi cell fractions were treated with exogenous protease and the percent of nascent apoAI and apoB accessible to proteolysis was determined and compared to that of albumin. At 5, 10, and 20 min of labeling, 35-56% of nascent apoAI and 60-75% of apoB in RER were degraded, while albumin was refractive to this treatment. At all times both apolipoproteins and albumin present in Golgi cell fractions were protected from proteolysis. These biochemical and morphological findings indicate that apoAI and apoB are associated with the rough microsomal membrane and are partially exposed to the cytoplasmic surface at early stages of secretion. They may later enter the luminal side of the ER and, on entering the Golgi, form lipoprotein particles.