A histochemical and ultrastructural study of yucca seed proteins

Embryos and nutritive tissues in ungerminated Yucca seeds of 4 species contain many spherical bodies which stain positively for proteins. Two distinct morphological types were observed at both the light and electron microscope levels. A meshwork-type consists of electron-dense and electron-transparent regions in which are embedded slightly birefringent inclusions. The second type, named the core-type, consists of a core surrounded by a matrix in which the inclusions are embedded. A single unit membrane surrounds each protein body. Both types are present in the embryo while only the core-type protein body appears in the surrounding nutritive tissue (perisperm). All regions in each of the two protein body types, except the inclusions, stain histochemically for proteins. Seeds were planted at 2-day intervals and allowed to germinate through 14 days. As germination commences (day 0) protein bodies in the embryo begin to break down. By day 4 the bodies are depleted in embryos of 3 of the 4 species. About day 4, protein bodies in perisperm surrounding the embryo begin to break down and this process continues outward to the seed coat until day 14 when all seed proteins have disappeared. During germination the protein bodies in the embryo and perisperm of 3 species coalesce and then undergo breakdown. In a fourth species, there is no appreciable increase in size of the bodies, but an erosion of the periphery and possibly internally as well takes place, followed by ultimate dissolution.     

Evidence for the presence of two different types of protein bodies in wheat endosperm

Storage proteins of wheat grains (Triticum L. em Thell) are deposited in protein bodies inside vacuoles. However, the subcellular sites and mechanisms of their aggregation into protein bodies are not clear. In the present report, we provide evidence for two different types of protein bodies, low- and high-density types that accumulate concurrently and independently in developing wheat endosperm cells. Gliadins were present in both types of protein bodies, whereas the high molecular weight glutenins were localized mainly in the dense ones. Pulse-chase experiments verified that the dense protein bodies were not formed by a gradual increase in density but, presumably, by a distinct, quick process of storage protein aggregation. Subcellular fractionation and electron microscopy studies revealed that the wheat homolog of immunoglobulin heavy-chain-binding protein, an endoplasmic reticulum-resident protein, was present within the dense protein bodies, implying that these were formed by aggregation of storage proteins within the endoplasmic reticulum. The present results suggest that a large part of wheat storage proteins aggregate into protein bodies within the rough endoplasmic reticulum. Because these protein bodies are too large to enter the Golgi, they are likely to be transported directly to vacuoles. This route may operate in concert with the known Golgi-mediated transport to vacuoles in which the storage proteins apparently condense into protein bodies at a postendoplasmic reticulum location. Our results further suggest that although gliadins are transported by either one of these routes, the high molecular weight glutenins use only the Golgi bypass route.     

Origin of nucleolus-like bodies found in the nucleoplasm and cytoplasm of Vicia faba meristematic cells

Cytohistochemical staining and RNase-gold labelling have been applied to root-tip meristematic cells of Vicia faba to study the origin and biological significance of 2 types of inclusions: one seen in the nucleoplasm and the other in the cytoplasm of early telophase cells. They have been termed "dense bodies" and "cytoplasmic nucleolus-like bodies" (NLB), respectively. Both types of inclusions respond positively to silver staining and ribonucleoprotein (RNP) staining in a similar fashion to nucleolus. Interestingly, the dense bodies label heavily with the RNase-gold complex, as does the nucleolus, while the cytoplasmic NLB have no affinity with the label. In most cases, the dense bodies label more heavily than the nucleolus. Light microscope surveys reveal that the dense bodies sometimes appear to be released from the surface of the nucleolus. On the other hand, prenucleolar material showing the same silver staining and RNP preferential staining characteristics as the dense bodies begin to accumulate on the surface of chromosomes in mid-anaphase. This material does not label with RNase-gold. These data are discussed in terms of the hypothesis that the dense bodies are derived from the nucleolus by direct budding or fragmentation, and the cytoplasmic NLB are composed of prenucleolar material that failed to attach to chromosomes.     

The fine structure of blood cells in the ascidian Perophora viridis

The fine structure of each of the blood cell types of Perophora viridis has been characterized and strong evidence for localization of vanadium in two of these types is given. There are eight cell types; phagocytes which may contain completely engulfed cells, lymphocytes with a prominant nucleolus and scanty cytoplasm packed with clustered ribosomes, and six other cell types each with distinctive granules. Morula cells contain a central nucleus and cytoplasm filled by wedged bodies, about five of which are seen in section. These bodies contain regularly spaced electron dense foci. Green cells have the same organization but contain bodies which are electron dense throughout. Granular amoebocytes contain many smaller lightly staining oval bodies and much glycogen. Another cell type (probably orange cells of light microscopy) contains numerous granular rounded bodies. Compartment cells have vacuoles containing electron dense particles and signet ring cells have usually one large vacuole which is electron dense lined and may contain electron dense particles. Developmental stages of these cell types show involvement of endoplasmic reticulum and Golgi bodies in granule formation. After glutaraldehyde fixation alone the only extremely electron dense components are particles in the compartment cells and signet ring cells implicating these as sites of vanadium localization, although not excluding other cell types.     

Water mites (Acari,Hydrachnidia) of water bodies of the Krąpiel River valley: interactions in the spatial arrangement of a river valley

The present study is a discussion of the interactions between different types of water bodies in the spatial arrangement of a river valley, taking into account landscape data. The Hydrachnidia assemblages in particular types of valley water bodies (oxbows, riparian pools, permanent ponds, flooded alder carrs, sedge marshes, and springs) are strongly influenced by the spatial arrangement of the water bodies in the landscape. Moreover, the formation of a fauna in a particular type of valley water body is also influenced by its origin. For example, the faunas of the oxbow lakes and riparian pools would have many characteristics in common, as these two types of water body can be characterized as originating in the river. As many as 61 species common to the valley water bodies and the Kr?piel River were noted. In the interactions between the valley water bodies and the river, the direction of migration from the former to the latter was clearly predominant. Migration in the reverse direction, from the river to the valley water bodies, took place to a far lesser degree. CCA analysis of landscape variables showed the influences of certain landscape parameters on water mites. These should be regarded as indirect influences, but as a consequence of their effects, they influence the formation of specific types of Hydrachnidia assemblages.     

The structure and cytochemistry of the oocytes in the crab Xantho bidentatus Milne Edwards

In the development of the oocytes of xantho bidentatus four stages could be distinguished. In stage I the cytoplasm is homogenous, in state II a perinuclear ring is formed, in stage III oocytes round bodies which are carbohydrate-protein complexes appear near the peripheri. These bodies occupy the oocyte completely in the stage IV oocyte. There are two types of bodies in the oocyte, big oval or round bodies which are carbohydrate-protein complexes and smaller bodies in between the oval bodies. These smaller bodies are lipid bodies. In stage I and II the cytoplasm is rich in RNA and in stages III and IV the cytoplasm is full of carbohydrates, proteins and lipids.     

Morphological criteria for the in vitro differentiation of embryoid bodies produced by a transplantable teratoma of mice

Embryoid bodies are produced when a transplantable testicular teratoma from strain 129 mice is serially passaged in the peritoneal cavity of these mice. These bodies are roughly spherical containing two morphologically distinct cell types. Scanning and transmission electron microscopy have been employed to show that the endodermal cells of embryoid bodies, like those of the mouse embryo, are on the outer surface and have highly convoluted surfaces containing numerous microvilli-like projections. The inner, embryonal carcinoma cells, are the pluripotent stem cells of this tumor. Intracisternal A-type particles have been observed in electron micrographs and are almost exclusively located in the endodermal cells of the embryoid bodies. The A-type complement-fixing antigen has been identified in extracts prepared from this tumor. When embryoid bodies are placed in culture and allowed to attach to the surface of a petri dish, a large number of new morphologically distinct cell types appear. Attachment to the petri dish surface is required for the generation of these new cell types. Cells of similar morphology in culture, display a distinctly “clonal” distribution on the petri dish surface.     

水稻叶片硅体与稻壳硅体的物相和光学性质比较

研究了水稻成熟叶片和稻壳中硅体的物相、自发荧光、红外和紫外吸收特性。X-射线衍射结果和显微红外摄谱结果一致表明稻壳硅体结构单一,为典型无定型矿质S iO2。稻壳硅体在紫外激发下没有自发荧光,表明稻壳硅体不含酚类化合物,因此硅体在285 nm处的强烈吸收为稻壳硅体本身所为。叶片硅体结构变异较大,硅体与标准物质无定形硅矿质之间存在细微的差异,并且哑铃形硅体、扇形硅体和不规则硅体的红外吸收光谱也不尽相同,叶片中这3种硅体在紫外激发下都能够自发荧光,表明叶片硅体中含有酚类化合物。硅体混合物仅在290 nm处有微弱的吸收,显示出叶片硅体在结构和性质上的变异性和复杂性。     

Studies on secretory activity in the pars intermedia of Xenopus laevis 2: A biochemical and electron cytochemical investigation of acid hydrolase activity following the stimulation of secretory activity in vivo

In the MSH cell at the onset of secretory activity, acid hydrolase activity increases. This increased activity, shown quantitatively by assaying beta-glycerophosphatase and R-glucuronidase within the stimulated gland, has been shown by electron cytochemical methods for beta-glycerophosphatase (acid phosphatase) and aryl sulphatase to be related to the production of large numbers of dense bodies. Cytochemical evidence also supports the view that these lytic bodies arise from GERL-like cisternal elements since it is shown that in addition to the flattened, parallel Golgi cisternae these elements are also R-glycerophosphatase-positive. The similarities between the dense bodies and those of other cell types are described and discussed.     

Modes of the formation of elementary bodies and their isolation from the cell in L-form bacteria

Elementary bodies are formed on the cell surface and inside the cell body in all cell types characteristic of L-form cultures, i. e. spherical cells, large bodies and filament structures. The following ways of elementary body formation are described: by budding on the cell surface, appearance immediately in the cytoplasm, in the vacuole, as a result of cytoplasmic fragmentation accompanied by the lysis of the cell, as well as in cases of the separation of cytoplasmic areas surrounded by the membrane or the myelin-like structure. The release of elementary bodies from the cell occurs as a result of the lysis or death of the mother cell, the thinning of the vacuole wall, and possibly due to small transient defects in the membrane, not accompanied by the death of the mother cell. The scheme of the formation and release of elementary bodies from the cell is presented.     

Protein Bodies at Early Stages of Development in High Lysine Mutant Notch-2 and Parent NP-113 Barley

In barley parent NP-113, endospermic protein bodies originate on rough endoplasmic reticulum, either as electron transluscent vesicles or as very small, spherical, electron dense protein bodies, These are translocated to vacuoles tor enlargement and subsequent storage, Endospermic protein bodies of Notch-2 high lysine mutant are either vacuolar, or confined to distended cisternae of smooth endoplasmic reticulum. Vacuolar protein bodies are of two types one, flocculent, which loosely fill up almost the entire vacuolar space; two, spherical, relatively compact and granular, Protein bodies, confined to smooth endoplasmic reticulum are small, spherical, electron dense or electron transluscent, These protein bodies fuse to form electron dense proteinaceous masses which are deposited in the cytosol due to disruption of the confining smooth endoplasmic reticulum.     

Ultrastructure of the testicular germinal zone and of the secondary spermatogonia in normal and destalked brachyuran decapod crustacea

Ultrastructural study of gonia of various species of male crabs shows that these cells belong to two types : primary and secondary gonia; these are discriminated according to their location inside or outside germinative islets, i. e. by the presence or absence of surrounding somatic tissue. The most original feature of the primary spermatogonia concerns the extrusion of gonial material under the shape of lamellar bodies or granules engulfed by the contiguous mesodermic cells; the latter degenerate when they are full of germ cell remnants. Poorly differentiated, the primary gonia have no great variety of cytoplasmic organelles. However two characteristic structures can be recognized in juxtanuclear position : nuage material (electron dense bodies more or less associated with gonia) and chromatoid bodies. Lipid droplets, the significance of which is unknown occur sometimes. Secondary spermatogonia divide synchronously although they are placed side by side without any connection such as cytoplasmic bridges. No ultrastructural modification has been noticed among the two types of gonia in crabs deprived of eyestalks. Mesodermic tissue distributed between primary gonia probably has a double role : inhibition of the onset of gametogenesis (as in Amphipods) and prevention of gonia cytolysis by collecting of altered elements.     

Nucleoli and perinuclear bodies in trophocytes of Panorpa communis contain factors of pre-mRNA splicing

The distribution of pre-mRNA splicing factors and protein coilin was examined in trophocyte nuclei (TN) in polytrophic ovarioles of Panorpa communis. In situ hybridization, using antisense U1 and U6 snRNA 3H-riboprobes, showed that TN were labeled evenly. Immunostaining at light and electron microscopic levels revealed in some TN nucleolar structures containing small nuclear RNP (snRNP) and protein coilin characteristic of the Cajal bodies/coiled bodies (CB). No free CBs were found in TN. These data showed that CB in TN are present only in the nucleoli. One of characteristic features of P. communis trophocytes is the presence of several types of perinuclear bodies (PB) in the cytoplasm. We distinguish between three types of PBs. PB-1 consist of spherical bodies (10-20 microns) with vacuoles composed of closely packed fibrils. PB-2 are irregularly shaped bodies (0.3-2.0 microns) consisting of a fibro-granular material. PB-2 are located near the nuclear envelope and contact the nucleoplasm material through nuclear pores. PB-1 and PB-2 join together to form a complex PB of the third type. All types of PB are not surrounded with a membrane and sometimes have mitochondria on their surface. The immunogold technique at the ultrastructural level revealed snRNP in PB-2. These results have enabled us to make a conclusion that PB-2 may be storage sites of snRNPs required for a future development of the embryo.     

Lamellar bodies are the intracellular site of membrane turnover in insect fat body

Analysis of the time course of highly cationic ferritin uptake by fat body cells has shown that the tracer bound to the plasma membrane and was pinocytosed by coated vesicles. The first sites of intracellular accumulation were multivesicular bodies which became filled with ferritin between 30-60 min after cells were exposed to the tracer. At no time during the experiments were any parts of the Golgi complex labeled by the tracer. By 60 min, the ferritin was increasingly found in lamellar bodies. The different types of 'light' and 'dark' multivesicular bodies suggest that lamellar bodies form from multivesicular bodies as they fill with tracer. The occurrence of lamellar bodies in many different cell types suggests an important role in membrane dynamics.     

包含体——流行性出血热病毒形态学的重要特征

用普通电镜和免疫酶电镜方法观察了流行性出血热病毒A9株和R22株感染的VeroE-6细胞中的病毒包含体,发现这些包含体具有以下三种基本形态:1.丝状或丝状颗粒包含体,2.松散颗粒包含体,3.致密包含体。在免疫酶法中,包含体均呈特异性酶染色阳性反应。描述和讨论了包含体与细胞及病毒发育成熟的关系。     

Blood cells and coelomocytes of the inarticulate brachiopod Lingula anatina

Three cell types are described from the coelomic cavity of the pedicle of the brachiopod Lingula anatina . Erythrocytes are abundant in the blood vessels of the mantle and also occur, in reduced numbers, in the pedicle. Phagocytic amoebocytes, characterized by a variable number of electrondense, homogeneous granules are common in the coelomic fluid of the pedicle. The enigmatic spindle bodies described by earlier authors constitute the most common cell type encountered in the pedicle coelom of aquarium-maintained specimens. The origin of spindle bodies from muscle cells is suggested.     

Evaluation of Escherichia coli cell disruption and inclusion body release using nucleic acid binding fluorochromes and flow cytometry

Many types of commercially valuable recombinant proteins produced by fermentation are expressed at high levels in Escherichia coli. Often, high-level expression in the host results in the formation of insoluble inclusion bodies. The release of these intracellular inclusion bodies from E. coli following cell disruption is a requirement for further downstream recovery. The ability to discern between intact unruptured cells and granules released from broken cells can provide valuable information for improving recovery yields in downstream purification. This paper describes a rapid and sensitive cytometry-based method that allows the simultaneous measurement of intact heat-killed E. coli and inclusion bodies using staining with nucleic acid binding fluorochromes.     

Ultrastructure, Origin, and Composition of the Protein Bodies in the Ligule of Isoetes lacustris L.

The basal cells in the ligule of Isoetes lacustris contain numerousprotein bodies, the contents of which can be digested enzymicallyby pronase and are stained red by treatment with ninhydrin Schiff'sreagent. Two types of protein bodies can be distinguished ultrastructurally:spherically-shaped bodies with granular contents and spindle-likebodies with fibrillar contents. Both are ensheathed by singlemembranes and do not show any solid inclusions within theirmatrix. The protein bodies probably arise from dilatation of the endoplasmicreticulum (ER) cisternae. This conclusion is based upon threeobservations: (a) The protein bodies occasionally show membranecontinuity with the ER; (b) ribosomes and polysomes are frequentlyattached to the protein-body membranes; (c) the contents ofthe protein bodies and of the dilated ER cisternae show similarultrastructural features. The dilatation of the ER cisternae is assumed to be a resultof protein accumulation in the intracisternal space. Based upon the results of polyacrylamide gel electrophoresis,it is likely that the spherically-shaped protein bodies storepredominately two proteins with molecular weights of 51300 and55800 D, while the spindle-like bodies store two proteins withmolecular weights of 92000 and 98000 D. The results presented do not permit a definite conclusion regardingthe function of the ligule of Isoetes lacustris but it is suggestedthat it may have a nutritive role. Isoetes lacustris L., ligule, protein bodies, endoplasmic reticulum, ultrastructure     

Biochemical criteria for the in vitro differentiation of embryoid bodies produced by a transplantable teratoma of mice. The production of acetylcholine esterase and creatine phosphokinase by teratoma cells

When embryoid bodies are grown in suspension culture in vitro, they undergo only a limited amount of morphological development. When these same embryoid bodies are permitted to attach to the surface of a culture dish, a wide variety of new morphological cell types appear. Suspension cultures of embryoid bodies do not contain significant detectable levels of acetylcholine esterase or creatine phosphokinase. These same enzymes however are produced in cell cultures derived from embryoid bodies attached to the culture dish surface. Polyacrylamide gel electrophoresis has been employed to demonstrate that the electrophoretic form of creatine phosphokinase produced by teratoma cells in culture is the brain form of the enzyme. Solid transplantable tumors containing only embryonal carcinoma cells (stem cells) do not contain either of these enzymatic activities. Well differentiated transplantable teratomas contain both enzymes.