Segregation of the Qb‐SNAREs GS27 and GS28 into Golgi Vesicles Regulates Intra‐Golgi Transport

The Golgi apparatus is the main glycosylation and sorting station along the secretory pathway. Its structure includes the Golgi vesicles, which are depleted of anterograde cargo, and also of at least some Golgi‐resident proteins. The role of Golgi vesicles remains unclear. Here, we show that Golgi vesicles are enriched in the Qb‐SNAREs GS27 (membrin) and GS28 (GOS‐28), and depleted of nucleotide sugar transporters. A block of intra‐Golgi transport leads to accumulation of Golgi vesicles and partitioning of GS27 and GS28 into these vesicles. Conversely, active intra‐Golgi transport induces fusion of these vesicles with the Golgi cisternae, delivering GS27 and GS28 to these cisternae. In an in vitro assay based on a donor compartment that lacks UDP‐galactose translocase (a sugar transporter), the segregation of Golgi vesicles from isolated Golgi membranes inhibits intra‐Golgi transport; re‐addition of isolated Golgi vesicles devoid of UDP‐galactose translocase obtained from normal cells restores intra‐Golgi transport. We conclude that this activity is due to the presence of GS27 and GS28 in the Golgi vesicles, rather than the sugar transporter. Furthermore, there is an inverse correlation between the number of Golgi vesicles and the number of inter‐cisternal connections under different experimental conditions. Finally, a rapid block of the formation of vesicles via COPI through degradation of ϵCOP accelerates the cis‐to‐trans delivery of VSVG. These data suggest that Golgi vesicles, presumably with COPI, serve to inhibit intra‐Golgi transport by the extraction of GS27 and GS28 from the Golgi cisternae, which blocks the formation of inter‐cisternal connections .     

Fine structure of actinorhiza of theRhamnaceae growing in Chile I.Talguenea quinquenervia (Gill et Hook)

Summary Root nodulesTalguenea quinquenervia Gill et Hook (Rhamnaceae) are restricted to the middle region of the root cortex. The root endophyte possesses hyphae which are septate and vesicles. The vesicles are spherical and are continuous with that of the hyphae. The endophyte fine structure is similar to otherFrankia-induced root nodules.     

Development,structure, and occurrence of secretory trichomes ofPharbitis

Summary Secretory trichomes develop from epidermal cells on the leaf primordia and stem ofPharbitis nil. Following an initial growth phase, trichomes begin active secretion of a protein-carbohydrate mucilage. This mucilage covers the shoot apex and developing leaves ofPharbitis.The secretory cells possess cellular organelles in forms usually associated with actively secreting cells: many mitochondria, an elaborate network of rough endoplasmic reticulum (RER), many free ribosomes, and numerous dictyosomes. The role of the dictyosomes is twofold: 1. dictyosome vesicles bud coated vesicles which transport materials from the cell and, 2. dictyosome vesicles coalesce, forming large storage vesicles. The storage vesicles are surrounded by, and often in contact with, poculiform RER. The RER forms an interconnected network throughout the cytoplasm, extending from the nuclear envelope to the plasmalemma. Distended profiles of RER are frequently in direct contact with the plasmalemma. Thus, inPharbitis secretory trichomes, it is the coated vesicles and RER which are active in secretion export. These findings imply a secretory pathway which deviates from the usual pattern in glandular cells.Predoctoral fellow of National Science Foundation during part of the investigation.     

Secretory organelle biogenesis: Trichocyst formation in a ciliated protozoan

Summary— By classical electron microscopy and immunoelectron microscopy, the biogenesis of trichocyst secretory granules has been followed in the ciliated protozoan Pseudomicrothorax dubius. The very early pre-trichocysts form by fusion of bristle-coated, electron-dense vesicles (dense vesicles) with electron-translucent vesicles (clear vesicles), both of which originate in a well-developed trans-Golgi network (TGN). The pre-trichocyst grows by further fusion with dense and clear vesicles as well as with other pre-trichocysts until it reaches its maximum diameter of about 2 μm. Dense and clear vesicle formation from the TGN has been followed, and the fusion sequence of dense vesicles with the pre-trichocyst has been documented. The contents of the dense vesicles are the precursors of the trichocyst tip, which is composed of four arm-like rods, whereas the shaft precursors are supplied by the clear vesicles. The first evidence of trichocyst shaft formation is the appearance of a paracrystalline, dense core condensation center in the pre-trichocyst. Following shaft formation, the trichocyst tip forms by fusion and condensation of the dense arm precursors along each of the four sides of the shaft. Docking of the fully formed trichocyst in the cell cortex is described. Pre-trichocyst biogenesis in cells grown with and without Se is compared.     

The ultrastructure of the innervation of the intestinal wall in the teleosts Myoxocephalus scorpius and Pleuronectes platessa

Summary The innervation of the intestinal wall in the teleosts Myoxocephalus and Pleuronectes was examined electron microscopically. Two classes of axons can be identified. The first, which is in the majority, contains numerous 50–150 nm granular vesicles as well as some 40–50 nm agranular vesicles while the second contains predominantly the 40–50 nm agranular vesicles. Chromate/dichromate staining methods suggest that the first type is aminergic. Both types lie in close association with the perikarya of intrinsic myenteric neurons but only axons containing predominantly agranular vesicles have synaptic membrane specialisations. No axon bundles pass into the longitudinal muscle layer in Myoxocephalus gut and though some do in Pleuronectes, they do not closely approach the smooth muscle cells. Axons containing large granular vesicles lie in intimate contact with the myocytes of the circular muscle layer. Both axon types pass through the submucosa to form a plexus underneath the mucosal epithelium. Varicosities containing agranular or granular vesicles are separated from the epithelial cells by a gap of about 200 nm in which lies a basal lamina.     

Pearl millet downy mildew (Sclerospora graminicola): Zoosporogenesis

Summary Cleavage of the zoosporangial cytoplasm ofSclerospora graminicola, the causal agent of pearl millet downy mildew, is by means of the fusion of cleavage vesicles and vesicles containing the extruded axoneme with the cell membrane. This type of zoosporogenesis linksS. graminicola to other Peronosporalean species, and is very similar to that seen for all uniflagellate species examined to date, while it separates it from species of theSaprolegniales where zoosporogenesis is brought about by the expansion of the central vacuole, or where the plasmalemma alone is used.The origin of the cleavage vesicles appears to be from the dictyosomes and not from the finger-print bodies which are rapidly formed in large numbers after axoneme formation and after the cleavage. vesicles have started to appear in the cytoplasm.     

ULTRASTRUCTURAL CHARACTERISTICS OF SPORE GERMINATION IN THE MOSS DAWSONIA SUPERBA

The spore wall of Dawsonia superba has characteristics that, in many respects, are similar to those of other mosses except for the exine, which is layered in Dawsonia. Imbibed spores have a well-developed endoplasmic reticulum with dilated cisternae that are associated with vesicles at the periphery of the cell. Ribosomes on the surface of the vesicles suggest that many vesicles originate from the endoplasmic reticulum. Two types of protein storage bodies are observed: membrane bound protein bodies with a homogeneous matrix which gradually becomes vesicular, and densely stained and non-membrane bound bodies consisting of crystalline arrays of fibrils. As in spores of higher plants, the protein reserves disappear during germination and may be converted to starch and other materials needed for development of the gametophyte.     

Histological description of the ink gland of the tropical sea hare Aplysia dactylomela Rang, 1828

This work describes for the first time the ink gland of Aplysia dactylomela Rang, 1828 using light microscopy and histochemical tests. The results reveal that this organ is covered by a layer of simple epithelium and that there are some differences between the epithelium facing the mantle shelf and that facing the mantle cavity. The former consists of columnar cells, which may have a secretory function, whereas the latter is a cuboidal epithelium. Underneath the epithelium and along the whole gland there are fibres of smooth muscle and collagen, organized in groups of parallel bundles. There are vesicles of different diameters, apparently with similar morphology. Some are filled with ink, whereas others are either granular or clear. The ink is released through a duct formed by invaginations of the cuboidal epithelium. Tests with bromophenol blue and periodic acid Schiff indicated that the ink and granulated vesicles contain proteins and carbohydrates or maybe glycoproteins. Between the fibre bundles and the vesicles there are dispersed cells. A diagram is presented emphasizing the covering epithelium, the distribution of muscle and collagen fibres, the dispersed cells, vesicles and ducts. This organization is similar to that of the other Aplysia species studied to date, A. californica.     

用于超声造影的微囊藻气囊提取新方法

气囊是一种由蛋白质外壳包裹气体形成的纳米级细胞结构,常见于蓝藻和嗜盐古菌中.气囊中包含的气体在超声波作用下,能够散射声波并产生谐波信号而增强信噪比,具备成为新型超声造影剂的潜质.但目前提取气囊的方法主要是传统的高渗裂解法,该操作过程烦琐、得率低,不适用于气囊的大规模提取.针对这些技术瓶颈,文中建立了一种快速、高效的微囊...     

Brush-border formation in the midgut of an insect,Calliphora erythrocephala meigen

Summary The embryonic development of the brush-border of anterior midgut cells of Calliphora was studied by electron microscopy. Dense surface-forming vesicles, as described by Bonneville (1970), are found prior to microvillus formation. These dense vesicles provide membranous and coating material for the moulding of the microvilli. The number of dense vesicles increases rapidly to a maximum just before brush-border formation, after which it decreases very rapidly, accompanied by an increase in the number of microvilli. Formation of microvilli proceeds in essentially the same way as in Xenopus. First, some of the vesicles fuse with the apical cell membrane, resulting in an increase of the cell surface, part of which is coated with filamentous material deriving from the dense vesicles. This in turn leads to bulging, and short irregular microvilli appear. These are erected and elongated.Prefabricated tubular elements are believed to play a part in this erection and elongation, probably due to the unwinding of spirally coiled strands.Microvillus formation proper lasts 2 to 3 hours in Calliphora. Almost the entire amount of membranous and coating material is prefabricated prior to the formation of microvilli.     

Pigment containing lipid vesicles

Summary Vesicles obtained by sonication of chlorophylla-lecithin mixtures dispersed in an aqueous medium closely resemble the well-characterized vesicles similarly prepared from pure lipids. They are bounded by one spherical lipid bilayer which contains the chlorophylla. Appropriate conditions for sonication prevent substantial degradation of the membrane constituents. Up to one chlorophylla molecule per 55 lecithins can be incorporated into the membranes. The average Stokes' radius of the vesicles determined by analytical sieve chromatography is 102±5 Å and independent of the chlorophylla content. The membrane is visible in the electron-microscope when the vesicles are treated with osmium tetroxide prior to negative staining. The osmium fixation is, however, not strong enough to allow for a preparation of the vesicles for thin sectioning (dehydration, embedding in epoxide).     

Ultrastructure of the vegetative organisation and initial stages of silica plate deposition in the soil testate amoebaCorythion dubium

Summary The siliceous body plates ofCorythion dubium are bound by a band of organic cement which is thickest at the lateral margins. The anterior vacuolar cytoplasm is separated by a pigment zone, which forms a dark band in the mid-body region, from the compact posterior region containing a typical vesicular nucleus surrounded by a region of dense endoplasmic reticulum. A pellicular basket of microtubules surrounds the posterior cytoplasm. The large Golgi complex lies between the nucleus and the fundus. Numerous coated and uncoated vesicles from the Golgi cisternae are seen in the peripheral cytoplasm alongside developing plates. These small siliceous plates are enclosed in silicon deposition vesicles lying in surface ruffles of the plasmalemma, often in association with a pair of microtubules. Observations are made on the formation of these vesicles and the early stages of silica deposition. A comparison is drawn between silica deposition inC. dubium and choanoflagellates where there is a similar association between silicon deposition vesicles and microtubules.     

Observations on the origin of two substances produced by the vitellaria in Gorgoderina attenuata

The vitelline cells in Gorgoderina attenuata produce two qualitatively distinct substances. One substance assumes the form of individual, dense, osmiophilic globules. Many globules are contained in a single vesicle. The other substance is an amorphous mass of medium density that completely fills a vesicle. Observations indicate that the dense, osmiophilic globules develop in association with a system of small, contiguous, ribosome-free vesicles. It is suggested that this system of vesicles constitute a Golgi apparatus for these cells. The amorphous mass substance develops in vesicles which appear to be derived from endoplasmic reticulum. Close association between the amorphous mass vesicle and mitochondria are commonly observed.     

Eversible abdominal vesicles and some observations of the male reproductive system of the spoon wing lacewing Palmipenna (Neuroptera: Nemopteridae)

The anatomy and histology of the abdominal eversible vesicles and the male reproductive tract of the spoonwing lacewing Palmipenna (Neuroptera: Nemopteridae) have been examined. The eversible vesicles open as a pair of large bulbous sacs between tergites five and six, each folding into halves during retraction. They consist of highly pleated cuticle, beneath which are typical gland cells, each having a circular or oval end apparatus surrounded by closely packed microvilli. These communicate to the surface via cuticularized channels. In spite of considerable behavioral observations, male Palmipenna were never noted with everted vesicles. Even during mating trials, where females were presented to males in the field, the vesicles were never everted during the attempted copulation that ensued. Our observations indicate that mate attraction is mediated by the release of a female pheromone. The function of the eversible vesicles and their associated gland cells remains unknown, and their structure appears to be unique to the Nemopteridae. The reproductive tract is similar to that of other Neuroptera, consisting of a pair of five-lobed testes, a medium-to-large pair of seminal vesicles, and three pairs of accessory glands. The major accessory glands are surrounded by circular and longitudinal muscle, and are lined by an epithelium, the cells of which presumably secrete the amorphous rods of material always present in this pair of glands. The sperm in the seminal vesicles are elongate, with a pointed head and a 9 + 9 + 2 configuration in the flagellum. A single spermatophore, similar in shape to that described for other Neuroptera, was found occluding the bursa copulatrix of a teneral female. © 1994 Wiley-Liss, Inc.     

Presence of a partially-coated reticulum in angiosperms

Summary The form and distribution of partially-coated membrane systems and coated vesicles in the leaf glands ofPhaseolus and the root cortical cells ofZea were investigated. Partially-coated membranes exist as a reticulum which is either sparsely branched or extensively anastomosed. Coated vesicles and coated membrane regions may occur at all points that are in the vicinity of such a reticulum. Golgi stack membranes, though associated with the partially-coated reticulum in many cases, show no consistent orientation to it. Occasionally, the reticulum and the associated coated vesicles are located away from any Golgi stack. We examined other plant tissues from such species asAllium cepa, Beta vulgaris andNicotiana tabacum and the partially-coated reticulum was observed in all material. We suggest that membrane flow may be occurring from the partially-coated reticulum to either the Golgi stack or to other intracellular compartments.     

Ultrastructural Localization of Enzymes Involved in the Feeding Process in Plasmodium chabaudi and Babesia hylomysci1

In P. chabaudi, hemoglobin digestion occurs in two ways: micropinocytosis and cytostomal phagocytosis. Both mechanisms lead to the formation of digestive vesicles which evolve to pigment vesicles containing hemozoin crystals. We used ultrastructural enzyme cytochemistry to detect and localize endoarylamidase and aminopeptidase activity. In P. chabaudi, these two enzymes are at first detected at the level of cytoplasmic ribosomes. When pinocytic vesicles appear, enzyme activity is localized at the membrane of the newly formed vesicles. Then, the labelling extends to the vesicle contents where it becomes very prominent. In the late trophozoite, enzymatic activity decreases and is no longer detected. In B. hylomysci, no endoarylamidase activity can be detected. Aminopeptidase is noted in the cytoplasm, the labelling being heavier in the growing trophozoites than in the younger stages. No vesicles or pigment can be observed. We thus conclude that aminopeptidase or endoarylamidase are synthesized in the cytoplasm of P. chabaudi and migrate to the digestive vesicles where hemoglobin digestion occurs. We do not know whether Babesia degrades hemoglobin since it does not produce residual pigment. It could feed from small peptides or amino acids coming from or through the stroma of the red blood cell.     

Freeze-substitution improves the ultrastructural preservation of legume root hairs

The freeze-substitution method was applied toVicia hirsuta root hairs to test its effectiveness in improving preservation of the cell ultrastruture. Freeze-substitution almost certainly represents more faithfully the structure of the hair cell. A previously unreported ‘pyriformis’ vesicle is described. Also unique to freeze-substituted material are coated secretory vesicles; a smooth plasma membrane profile; mitochondrial ribosomes; long microfilament bundles which are associated with vesicles, mitochondria, coated pits and coated vesicles; microtubule-associated filaments; well-preserved coated vesicles and coated pits with enclosing rings; a pliciform nucleus. The results are discussed in context of previous reports using conventional fixation techniques.     

Hyphal structure in Fusarium oxysporum (Schlecht) revealed by freeze-etching

Summary Freeze-etched hyphae of F. oxysporum exhibited a single layered cell wall; a plasmalemma, in which invaginations were frequently associated with paramural vesicles; cytoplasma bearing lipid droplets, vacuoles, intravacuolar vesicles and nuclei with typical nuclear pores. Some hyphae bore crystalline inclusions characterized by a pronounced hexagonal, external ornamentation and it is suggested that the presence of these crystals and intravacuolar vesicles are indicative of aging hyphae.     

The golgi-mediated scale production in the marine haptophycean alga Ochrosphaera neapolitana Schussnig

Abstract

Some ultrastructural features of cells of the marine haptophycean alga, Ochrosphaera neapolitana Schussnig in the palmelloid stage were examined. Chloroplasts which are contained in a compartment isolated from the cytoplasm by ER profiles and nuclear envelope, display trilamellated thylakoids running along the major axis. The stalked pyrenoid with inner bilamellated thylakoids, protrudes in a large membrane-bounded vacuole. Other structures, as the haptonematic and flagellar bases, autophagic vacuoles and mitochondria, are typical of the chrysophycean and haptophycean genera so far investigated.

The Golgi apparatus is represented by a single dictyosome composed of stacked cisternae fonctioning in a way that they form organic scales which constitute the main part of the cell covering. The scales, build up of microfibrils disposed parallel each to other, lie in cisternal lumina of the dictyosomal maturing face; scaly cisternae are numerous in the peripheral cytoplasm and are observed merging in the plasma membrane and discharging the content outside the protoplast.

Dictyosomal activity is evidenced morphologically by massive vesicle production. Three kinds of membrane-bounded vesicles were identified in the present material: i) inner-granulated vesicles, arising from the maturation face; ii) coated vesicles, scattered in the cytoplasm or at the periphery of the golgi body, and iii) dense-cored vesicles, present in the proximity of the maturation face. The possible functional relationships related to scale production and assembly outside the protoplast, and between the nucleus and dictyosome are discussed.     

The properties and buoyancy-providing role of gas vacuoles in Trichodesmium Ehrenberg

All three species of the marine blue-green alga Trichodesmium collected in the Sargasso and Caribbean seas were found to possess gas vacuoles. The constituent gas vesicles were much stronger than those found in any freshwater blue-green alga, the mean critical collapse pressures being 12 bars in T. erythraeum, 34 bars in T. contortum and 37 bars in T. thiebautii. This great strength is obviously an adaptation to the hydrostatic pressures at the depths to which these organisms occur in the ocean. In each case the gas vesicles are far too strong to be collapsed by rising cell turgor pressure, though gas-vacuolation could be slowly regulated by the differential growth of gas vesicles and cells. Since the vesicles are of a similar shape and size to those in other species, the vesicle wall material must be stronger. The majority of Trichodesmium colonies collected were positively buoyant, and in all cases tested the buoyancy was dependent on the presence of gas vacuoles. The buoyancy is important in increasing the residence time of these slowly growing algae in the euphotic zone and it is responsible for the surface water-blooms which they form.