Distribution of golgi apparatus-associated polyribosomes across the polarity axis of dictyosomes of wild carrot (Daucus carota L.)

Summary Endoplasmic reticulum-polyribosome-Golgi apparatus associations were a general feature of cells of suspension cultures of wild carrot (Daucus carota L.). Free polyribosomes occurred within the Golgi apparatus zone for all dictyosomes and with equal frequency at all levels within the stack including the most mature or trans face. When evaluated and quantified from electron micrographs, approximately 60% of the dictyosome profiles were characterized by a system of transition elements consisting of part smooth-part rough endoplasmic reticulum. These were encountered most frequently in the immediate vicinity of the immature, forming or cis face, usually toward the periphery of the stacked cisternae. Analysis of serial sections showed that those dictyosome profiles not exhibiting this characteristic did so primarily because of an unfavorable plane of sectioning. All dictyosomes examined in 5 or more serial sections revealed some type of close association with endoplasmic reticulum. Some of the associations were so close that direct connections between Golgi apparatus and endoplasmic reticulum tubules could not be excluded. Also present, especially at the forming or cis face, were small 600 nm transition vesicles with nap-like surface coats on nearly 90% of the dictyosomes examined. More than 50% exhibited spiny (clathrin-)coated vesicles at the mature or trans face.     

Cytodifferentiation in the Rana pipiens oocyte

Summary In early diplotene frog oocytes incubated to illustrate thiamine pyrophosphatase (TPPase) activity, reaction product is uniformly distributed within the compartments of the endoplasmic reticulum and nuclear envelope as well as within the saccules and small vesicles comprising the dictyosomes. With continued oocyte development the reaction product becomes concentrated in localized regions of the dictyosome saccules. Eventually, the enzyme is no longer apparent within the endoplasmic reticulum, but is concentrated in the cisternae of the inner dictyosome saccules. The variations noted suggest that the enzyme is synthesized early in diplotene by the endoplasmic reticulum and is subsequently transported to the Golgi apparatus where it is consistently observed at later developmental stages. TPPase activity is also present in the Golgi apparatus of follicle and theca cells as well as in ovarian epithelial cells. The enzyme is also detected in micropinocytotic vesicles contained within the cells comprising the follicle envelope and in intercellular spaces of the follicle. Horseradish peroxidase injected into the coelomic cavity is transported via micropinocytotic vesicles into and through the cells comprising the follicle envelope and in intercellular spaces. The exogenous protein is not found even after a prolonged time period in early diplotene oocytes. The protein is, however, present in large spherical and tubular vesicles in the cortex of vitellogenic oocytes approximately 500 microns in diameter. The possible functional role of the enzyme TPPase during oogenesis is discussed.This investigation was supported by a research grant from the National Science Foundation (GB-8736).     

Development of compound trichocysts in the ciliatePseudomicrothorax dubius

Summary During the logarithmic growth of the ciliatePseudomicrothorax dubius associations between mitochondria, rough endoplasmic reticulum and dictyosomes have been observed. The Golgi apparatus is very active and it is suggested that, as a consequence of cytotic activity, the contents of the Golgi vesicles become incorporated into large irregular vacuoles as globular material. The large vacuoles develop into trichocysts and the dictyosome derived globules consolidate to ultimately form the rod-like arms of the trichocysts of theMicrothoracidae.     

CYTOCHALASIN B INFLUENCES DICTYOSOMAL VESICLE PRODUCTION AND MORPHOGENESIS IN THE DESMID EUASTRUM1

Cytochalasin B (CB) applied to young developing cells of the desmid Euastrum oblongum Ralfs ex Ralfs, at concentrations that do not entirely inhibit cytoplasmic streaming, retarded cell growth and caused malformations of cell shape. While the basic symmetry of the cell was maintained, only the first indentations were formed and the cell body appeared to be swollen. Electron microscopic investigations revealed that vesicle production at the dictyosomes was disturbed by cytochalasin. In contrast to untreated control cells, where vesicles with electron-dense contents (“dark vesicles”) were formed during primary wall formation, vesicles pinched off by the dictyosomes during CB treatment exhibited an “empty” appearance. These vesicles, which correspond to the “dark vesicles” in size, were accumulated around the dictyosomes without being transported to the plasma membrane and were frequently connected to the trans-cisternae of the Golgi bodies. We speculate that CB may influence the transfer of products from the endoplasmic reticulum (ER) to the dictyosomes via transition vesicles, which results in a disturbed vesicle production at the Golgi bodies. CB also causes a shift in ER and dictyosome distribution. Moreover, a cortical actin system appears to be involved in the cell shaping of Euastrum. The arrangement of microtubules around the nucleus is not affected by the drug.     

Intracellular transport of albumin through the secretory apparatus of rat liver parenchymal cells. An immunocytochemical study

The fine structural localization of albumin in rat liver parenchymal cells was determined by an improved immunocytochemical method and serial sectioning. Albumin in the secretory apparatus of the parenchymal cells was present in segments of the rough endoplasmic reticulum, interrupted with negative segments, in transport vesicles, Golgi saccules, finely anastomosed tubules and vesicles on the trans side of the Golgi complex, and in secretion granules. Horizontally sectioned Golgi saccules contained lipoprotein particles on one side and albumin on the other side. After transport, the vesicles that contained albumin fused with the so-called rigid lamellae on the trans-side of the Golgi complex. Ultrathin serial sections revealed no true structural continuity between the endoplasmic reticulum and the cis-aspect of the Golgi complex. We concluded that secretory proteins are transported from the endoplasmic reticulum to the Golgi complex by transport vesicles that bud from the endoplasmic reticulum and fuse with the Golgi saccules. These vesicles fuse regularly with the Golgi saccules on the cis-side and occasionally with tubular elements on the trans-aspect that may belong to the so-called GERL.     

Pentachlorophenol affects mitochondria and induces formation of golgi apparatus-endoplasmic reticulum hybrids in tobacco pollen tubes

Summary The toxic effect of pentachlorophenol (PCP) on the growth and ultrastructure of tobacco pollen tubes was tested using a semivivo technique of tube culture. In this technique the pollen tubes were allowed to grow in the pistilin situ for 24 hr before they protruded from the cut end of the style and came into contact with the medium containing PCP. The inhibitory effect of different PCP concentrations was determined by measuring the length of tube bundles. The intracellular action of PCP was analysed by electron microscopy. This biocide caused four obvious alterations in the pollen tube ultrastructure: (1) swelling of the mitochondrial saccules; (2) enlargement of the dictyosomes by the increase of the cisternal diameter and the number of cisternae per stack; (3) formation of cup-shaped Golgi apparatus-endoplasmic reticulum hybrid structures (GER hybrids) showing continuities of ER and Golgi cisternae; (4) formation of stacked and/or concentric arrangements of rough ER cisternae. It is suggested that swelling of saccules was directly due to the uncoupling of oxidative phosphorylation whereas the changes of the endomembrane system were caused by energy depletion due to the inhibition of ATP synthesis. These changes are consistant with dynamic concepts of dictyosome and ER function when membrane formation exceeds membrane use in the production of secretory and transition vesicles. Thus, the enlargement of the dictyosomes and the formation of GER hybrids are thought to result from inhibition of budding of vesicles from the Golgi apparatus or from both the ER and the Golgi apparatus, respectively.     

The endomembrane system and mechanism of membrane flow in the green alga,Gloeomonas kupfferi (Volvocales,Chlorophyta) I. An ultrastructural analysis

Summary The endomembrane system of the chlamydomonad flagellate,Gloeomonas kupfferi (Chlorophyta), is complex. It consists of a proliferating ER network, a perinuclear complex of 14–18 dictyosomes and 8–12 vacuoles and an anterior contractile vacuole complex. The ER network extends from the nuclear envelope outwards, ensheafhs a dictyosome, extends out through a lobe of the chloroplast and terminates in the thin zone of peripheral cytoplasm between the chloroplast and plasmamembrane. The individual dictyosome is polar with distinct cis- and trans-faces. The cis-face is closely associated with transition vesicles emerging from the adjacent ER. Large vesicles emerge from peripheral swellings of terminal cisternae. The dictyosome-associated ER is connected to the peripheral vacuolar system. During cell division and cytokinesis, changes in the endomembrane system occur. Dictyosomes divide and quickly separate to form perinuclear complexes around the daughter nuclei. Each dictyosome undergoes morphological changes during this wall precursor-producing stage. ER lines the furrow zone and is closely associated with phycoplast microtubules. A discussion of the endomembrane system in membrane flow mechanics is provided.Abbreviations ER endoplasmic reticulum - OsFeCN Osmium ferricyanide     

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.     

The Endomembranes ofchlamydomonas reinhardii: A comparison of the wildtype with the wall mutants CW 2 and CW 15

Summary Ultrastructural observations on the principal endomembranes (endoplasmic reticulum, Golgi apparatus) of synchronously growing of wild type and mutant (CW 2, CW 15) strains ofChlamydomonas reinhardii have been carried out. The dictyosomes of the Golgi apparatus in all three cases are highly polar in morphology but lack intercisternal filaments. A clear spatial relationship between dictyosomes and endoplasmic reticulum is seen and a transfer of vesicles from the latter to the former is easily visualized. Coated vesicles invariably appear to be restricted to the trans-pole of the dictyosomes. The endoplasmic reticulum adjacent to the cis pole of dictyosomes is considerably hypertrophied in the case of the wild type, only partially so in the mutant CW 2 but not at all in the mutant CW 15. In the wild type this swelling is most extreme during the period of wall deposition and for several hours afterwards. The results are discussed in relation to the biosynthesis and intracellular transport of, particularly O-glycosidically linked, glycoproteins.     

Selective staining of dictyosome-like structures (DLS) from spermatocytes of the guinea pig using phosphotungstic acid at low pH

Dictyosome-like structures (DLS) occur abundantly in primary spermatocytes of the guinea pig. DLS superficially resemble dictyosomes of Golgi apparatus in that they consist of stacked cisternae and react similarly to some cytochemical markers. DLS saccules are also present in residual bodies and in the cytoplasmic droplet of the sperm, but the stacked configuration (or dictyosome form) is seldom present at these stages of development. A mixture of 1% phosphotungstic acid in 10% chromic acid selectively stains the DLS and DLS saccules of guinea pig germ cells. The thick cisternae of spermatid Golgi apparatus and the sperm plasma membrane also stain, but endoplasmic reticulum and the parts of the Golgi apparatus other than the thick cisternae do not stain. The specificity of the stain is retained in crude homogenates as well as in purified cell fractions and may be helpful in identification of DLS in cell fractionation studies. Additionally, the information obtained provides clues to the origin and fate of DLS in the developing mammalian germ cells.     

Endoplasmic Reticulum-dictyosome Involvement in the Origin of Refractive Spherules in Sieve Elements of Davallia fijiensis Hook

Young sieve elements from petioles and rachises of Davalliafijiensis Hook were examined with an electron microscope. Evidencewas obtained that implicated both the endoplasmic reticulum(ER) and the Golgi apparatus in the formation of refractivespherules. Numerous connections were observed between smooth,tubular ER and peripheral tubules of the dictyosomes, indicatingthat these two cytoplasmic components are parts of a singleendomembrane system. Davallia fijiensis Hook, endomembrane system, endoplasmic reticulum, dictyosome, refractive spherule, sieve element     

The neural gland in tunicates: fine structure and intracellular distribution of phosphatases

Summary The cells comprising the neural gland in the ascidians Ciona, Styela, and Botryllus have been examined for their fine structural features and enzyme cytochemistry. The gland cells are either cuboidal or irregular in outline. They are full of small vesicles, of which some are pinocytotic, as well as larger vacuoles; they become increasingly vacuolated as their shape decreases in regularity. At the same time, glycogen deposits accumulate and the cisternae of the endoplasmic reticulum become distended. Some of the vacuoles contain an electron dense material or a fibrillar substance, but the cells contain no obvious electron opaque secretory granules associated with an extensive Golgi complex such as occur in the vertebrate adenohypophysis.Acid phosphatase is localized in some of the vesicles and vacuoles, indicating that they are a kind of lysosome, the latter possibly representing autophagic vacuoles. Thiamine pyrophosphatase is also found in many vacuoles as well as in the saccules of the Golgi apparatus which in these cells is in the form of dictyosomes.The results suggest a developmental cycle of increasing cytoplasmic vacuolation, ultimately leading to a breakdown and release of the vacuolar products. The significance of these observations is considered, particularly with respect to the hypothesis that the gland represents the ascidian equivalent of the vertebrate pituitary.I am grateful to Miss Yvonne R. Carter for technical assistance with the photography and to Mr. John Rodford for producing the diagram.     

Developmental and comparative ultrastructure of glandular hairs in the two Urticaceae. I. Urtica dioica

Secretion produced by glandular hairs is deposited mainly in the periplasmic space of the head cells. It stains intensely for both proteins and polysaccharides. The ultrastructure of meristematic, differentiating, mature and senescent head cells as well as the stalk and basal cells has been described in comparison to that in other cell types of the leaf. The specific features of the head cells are the proliferation of the granular endoplasmic reticulum as well as the multiplication of the dictyosomes and mitochondria during transition to the secretion stage. However, the frequency of dictyosomes varies among secreting hairs. The ER produces neither secretory nor transition vesicles and does not anastomose with the plasmalemma. In the absence of transition vesicles, the transport of secretory proteins and enzymes of polysaccharide synthesis from the ER to dictyosomes apparently includes the cytosolic step. Dictyosomes, though not appearing hypersecretory, produce two types of smooth secretory vesicles generated by the trans Golgi reticulum. The vectorial transfer of prosecretion and membranes across the dictyosome stack proceeds via the transport (shuttle) vesicles. It is, therefore, concluded that exocytosis of smooth secretory Golgi vesicles is the sole mechanism of release of both proteins and polysaccharides. Coated vesicles occasionally seen near the plasmalemma are likely to be involved in the endocytotic membrane retrieval. The secretion product disappears during senescence of the hairs and the secretory cells undergo vacuolation by means of local autophagy.     

The structure of the Golgi apparatus: a sperm’s eye view in principal epithelial cells of the rat epididymis

 The Golgi apparatus of epididymal principal cells shares many structural features with other cell types. Saccular regions are arranged in a cis-Golgi network, eight flattened saccules, and several trans-Golgi networks (TGNs). Dilated tubules form intersaccular connecting regions which joint together saccules at the same or different levels between adjacent stacks. Wells exist as large perforations in register with the four cis-most saccules and serve as areas of vesicular interactions. TGNs are variable and can appear to peel off the stack or to be detached from it in the form of an anastomotic tubular network with pale dilated areas corresponding to prosecretory granules connected by short narrow bridges. Elongated or discoid dilated cisternae of endoplasmic reticulum (ER) (sparsely granulated) lie over the cis face of the stack, from which they are separated by an intermediate compartment filled with vesicles and tubules. The ER is also closely juxtaposed to the TGNs and the eighth saccule but interconnections are never seen between them. Vesicles of the COP variety reside at all levels of the stack and appear to bud off the cis-located ER and the edges of the saccules, while clathrin-coated vesicles appear mainly on the trans face of the stack and next to lysosomes. In the supranuclear cytoplasm, clusters of vesicles and tubules, at times budding off enveloping ER, appear to radiate toward the Golgi stacks where they fuse with cis Golgi elements. Taken together, these observations suggest dynamic functions and interactions for the various Golgi elements, associated vesicles, ER, and vesicular tubular clusters. Accepted: 29 January 1998     

ULTRASTRUCTURE AND ACID PHOSPHATASE IN PEDICEL ABSCISSION OF HIBISCUS

Pedicel abscission in Hibiscus rosa-sinensis was investigated by light and electron microscopy. During the pre-abscission period endoplasmic reticulum declined somewhat, dictyosomes increased in number and apparent activity, and mitochondria maintained their numbers. The observations suggested that dictyosomal vesicles were migrating to and fusing with the plasma membrane. The enzyme acid phosphatase was associated with dictyosomes and dictyosomal saccules, with small vacuoles and invaginations of the plasma membrane, and in the paramural region between the plasma membrane and the cell wall. Our interpretation is that acid phosphatase, (and probably also the enzymes involved in cell wall dissolution) are transported via an endoplasmic reticulum-dictyosome-vesicle carrier system to the paramural regions of the cell. In more general terms, our observations support the view that the enzymes involved in the cell wall hydrolysis of abscission are synthesized within a compartmentalized, lysosomal system prior to their release and action.     

Further studies of the glandular tissue of the Sauromatum guttatum (Araceae) appendix

Electron microscopic studies showed that the trans-Golgi network (trans indicates the polarity of cisternae within the Golgi apparatus; it is opposite to the cis-face that is adjacent to the rough endoplasmic reticulum) was involved in the processing of the osmiophilic material present in the appendix of the inflorescence of Sauromatum guttatum. This material accumulated in the rough endoplasmic reticulum and in special pockets of the plasma membrane prior to heat production. Associations between the endoplasmic reticulum and trans-Golgi network were observed. The Golgi apparatus was composed of 5–6 dictyosomes on one side and one or two somewhat detached cisternae on the other side. Various nonosmiophilic Golgi-derived vesicles were observed: small ones covered with spike-like material, large ones with a smooth surface, and irregularly shaped ones. These electron-translucent vesicles seemed to accumulate in specific localities at the plasma membrane surface in the vicinity of the osmiophilic material; they were not found when the aroma was released. During heat production, the Golgi structures shrank and the activity of the trans-Golgi network seemed to be reduced. At the same time, coated pits were seen at the plasma membrane surface. In some cells, hypertrophic Golgi apparatuses were seen with only 2–3 dictyosomes that contained granulated material in their lumens. Finally, the osmiophilic material was also found in the plasmodesmata.     

Changes in the activity of the Golgi apparatus during the cell cycle in antheridial filaments ofChara vulgaris L.

Summary The number of dictyosomes found in one central cell section in antheridial filaments ofChara vulgaris increases proportionally to the cell length during interphase. The activity of Golgi apparatus was expressed by a number of Golgi vesicles surrounding a single dictyosome. These vesicles are most numerous during mitosis and cytokinesis,i.e., prior to and during cell plate formation. In the middle and late S phase the number of Golgi vesicles decreases by about 25%; subsequently, during the early and middle G₂, it increases again. At the end of the G₂ phase, Golgi vesicles are the scarcest.The increase in the number of Golgi vesicles during the G₂ phase coincides with the period of intense cellular elongation, and, thus, it is probably related to the enhanced synthesis of cell wall components.Coated vesicles are most numerous in prophase, metaphase, and early telophase, and during interphase in both late S and G₂ phase. It was found that the number of coated vesicles is proportional to the degree of condensation of nuclear chromatin.This work was supported by the Polish Academy of Sciences within the project 09.7.3.1.4.     

AN ULTRASTRUCTURAL BASIS FOR HYPHAL TIP GROWTH IN PYTHIUM ULTIMUM

Hyphae of the fungus Pythium ultimum extend by tip growth. The use of surface markers demonstrates that cell expansion is limited to the curved portion of the hyphal apex. Growing and non-growing regions are reflected in internal organization as detected by light and electron microscopy. The young hypha consists of three regions: an apical zone, a subapical zone and a zone of vacuolation. The apical zone is characterized by an accumulation of cytoplasmic vesicles, often to the exclusion of other organelles and ribosomes. Vesicle membranes are occasionally continuous with plasma membrane. The subapical zone is non-vacuolate and rich in a variety of protoplasmic components. Dictyosomes are positioned adjacent to endoplasmic reticulum or nuclear envelope, and vesicles occur at the peripheries of dictyosomes. A pattern of secretory vesicle formation by dictyosomes is described which accounts for the formation of hyphal tip vesicles. Farther from the hyphal apex the subapical zone merges into the zone of vacuolation. As hyphae age vacuolation increases, lipid accumulations appear, and the proportional volume of cytoplasm is reduced accordingly. The findings are integrated into a general hypothesis to explain the genesis and participation of cell components involved directly in hyphal tip growth: Membrane material from the endoplasmic reticulum is transferred to dictyosome cisternae by blebbing; cisternal membranes are transformed from ER-like to plasma membrane-like during cisternal maturation; secretory vesicles released from dictyosomes migrate to the hyphal apex, fuse with the plasma membrane, and liberate their contents into the wall region. This allows a plasma membrane increase at the hyphal apex equal to the membrane surface of the incorporated vesicles as well as a contribution of the vesicle contents to surface expansion.     

Connections between dictyosomes,ER and GERL in cotyledons of mung bean (Vigna radiata L.)

Summary The connections and structural inter-relations of dictyosomes and endoplasmic reticulum (ER) in cotyledons of germinating mung beans were studied using thick (0.3 m) sections of aldehyde fixed, zinc iodide-osmium tetroxide (ZIO) impregnated tissue. The sections were examined by conventional (100 kV), rather than high voltage, transmission electron microscopy.Continuity of cisternal ER with tubular ER was confirmed and a direct connection of tubular ER totrans dictyosome cisternae was observed as were GERL networks associated withtrans dictyosome cisternae.Dictyosomes also gave rise to an extensive system of very fine tubules (10–20 nm diam) which have not been described previously in plant tissue. These tubules, which originated at thetrans dictyosome face, extended throughout the cytoplasm and were found connected to cisternal ER and tubular ER.The implications of these observations are discussed with regard to present ideas concerning endomembrane flow and protein sorting by the Golgi apparatus.     

Hydrogenosome-Methanogen Assemblages in the Echinoid Endocommensal Plagiopylid Ciliates,Lechriopyla mystax Lynch, 1930 and Plagiopyla minuta Powers, 19331

ABSTRACT. Lechriopyla mystax Lynch, 1930 and Plagiopyla minula Powers, 1933 contain hydrogenosome-methanogen assemblages similar to those reported for other plagiopylid ciliates. These assemblages are stacks of elongate ovoid hydrogenosomes alternating with methanogens; these stacks are surrounded by cisternae of the rough endoplasmic reticulum that are often accompanied by Golgi complexes. The individual methanogens in the larger ciliate, L. mystax, are about four times the volume of those in the smaller ciliate, P. minuta, but both ciliates appear to contain Gram-negative methanococcoid bacteria, possibly Methanoplanus sp. The endoplasmic reticulum-Golgi complex probably plays a significant role in exploitation of the methanogens by the host ciliate.