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A Ellinger

Publications and source records attributed to A Ellinger.

At least 73 records · Page 4Linked to original sources

Localization of binding sites for concanavalin A, Ricinus communis I and Helix pomatia lectin in the Golgi apparatus of rat small intestinal absorptive cells.

Binding sites for concanvalin A (Con A), Ricinus communis I agglutinin (RCA I), and Helix pomatia lectin (HPA) were localized in the Golgi apparatus of rat small intestinal absorptive cells. A preembedment technique, a modification of the one originally used by Bernhard and Avrameas (Exp Cell Res 64:232, 1971), was employed, with horse-radish peroxidase being used for cytochemical visualization. Incubations were performed on 10 microns thick cryosections of duodenal segments that were fixed in a mixture of 4% formaldehyde and 0.5% glutaraldehyde; fixation was preceded by a 2-min rinse in 0.1 M sodium cacodylate and followed by storage in the same buffer for up to 7 days. Incubation with Con A, which binds preferably to alpha-D-mannose and alpha-D-glucose residues, caused intense reaction of the dilated cisternae of the cis Golgi side; staining was variable in intermediate and trans cisternae. RCA I, recognizing beta-D-galactose residues, could only be demonstrated in intermediate cisternae. Reaction for HPA, which indicates alpha-N-acetyl-D-galactosamine residues, stained intensely 1 to 2 cisternae of the cis Golgi side, as well as being localized in the peripheral regions of the cisternae of the intermediate compartment of the stacks. Deposits of reaction product covered the luminal surface of the cisternal membranes, but usually left the lumen itself, as well as lipid particles, devoid of reaction. The differences in Con A, RCA I, and HPA reactivity between cis, intermediate, and trans cisternae suggest compositional and structural differences of the carbohydrates in the respective compartments; they may reflect conversion processes that are known to occur in the oligosaccharide side chains of glycoconjugates at the Golgi complex level.

Absorption↗

Effect of monensin on the Golgi apparatus of absorptive cells in the small intestine of the rat. Morphological and cytochemical studies.

The effect of short-time treatment with the ionophore monensin, administered intraluminally at concentrations of 5 and 10 microM, was studied on the Golgi apparatus of absorptive cells in the small intestine of the rat. At 2-3 min after treatment most of the Golgi stacks exhibited dilated cisternae. At 4-5 min stacked cisternae were absent; they were replaced by groups of smooth-surfaced vacuoles. Dilatation and vacuolization occurred in the entire stacks without preferential effect on any particular Golgi subcompartment. Monensin did not influence the cytochemical Golgi reaction of thiamine pyrophosphatase and acid phosphatase. The characteristic staining pattern of these two enzymes in all Golgi cisternae of absorptive cells in the proximal small intestine, and the reactivity restricted to trans cisternae in distal segments of the small intestine, were unchanged after treatment with monensin. In the distal small intestine, the cytochemical pattern allowed the monensin-induced vacuoles to be attributed to the former cis- or trans-Golgi face. Further, the cytochemical results demonstrate that vacuolization is not restricted to the stacked cisternae, but includes the trans-most cisterna. The latter, usually located at some distance from the Golgi stacks, has been defined as belonging to the GERL system in several types of cells. The clear response to monensin, an agent that selectively affects the Golgi apparatus, indicates common properties between trans-most and stacked Golgi cisternae.

Acid Phosphatase↗

Colchicine-induced tubular, vesicular and cisternal organelle aggregates in absorptive cells of the small intestine of the rat. I. Morphology and phosphatase cytochemistry.

Treatment of rats with colchicine administered intraperitoneally at a dosage of 0.5 mg per 100 g of body weight for 6 hr induces extensive accumulations of tubular-vesicular and cisternal organelles in the absorptive cells of the small intestine. The formation of these organelle aggregates coincides with a reduction of microtubules and massive changes in the cellular organization including alterations of the Golgi apparatus and the plasma membrane. In most cases the accumulated tubules and vesicles contain a homogeneous electron-dense matrix, the cisternae often having the character of rigid lamellae. The organelle aggregates mainly occupy apical cell portions subjacent to the terminal web as well as basal cellular regions close to the basolateral plasma membrane. Tubular-vesicular as well as cisternal organelles react strongly for thiamine pyrophosphatase (TPPase), inosine diphosphatase (IDPase), acid phosphatase (AcPase) and trimetaphosphatase (TMPase). The staining pattern of TMPase differs from that of the other phosphatases in that the reaction is restricted to the colchicine-induced tubular-vesicular and cisternal aggregates, whereas TPPase, IDPase, and AcPase, respectively, also appear over Golgi stacks, multivesiculated bodies and plasma membrane. This phosphatase reactivity indicates the lysosomal character of the organelle aggregates.

Acid Anhydride Hydrolases↗

Effect of colchicine on rat small intestinal absorptive cells. I Formation of basolateral microvillus borders.

Treatment of rats with colchicine (0.5 mg/100 g of body weight) for more than 3 hr causes formation of microvillus borders along lateral and basal surfaces of absorptive cells in the small intestine. Morphologically, these strongly resemble the apical brush border inclusive of the terminal-web region. Formation of basolateral microvilli is restricted to mature absorptive cells. At 6 hr after administration of colchicine, 3.47% (+/- 1.94%) of the basolateral cell surfaces exhibit "implantation" of microvillus borders. The results show that colchicine induces formation of surface differentiations at lateral and basal surface regions that are restricted to the apical cell surface in controls. Redistribution of constituents of the plasma membrane from apical to basolateral membrane portions, as well as rearrangement in the organization of microfilaments can be considered to underlie formation of basolateral microvillus borders. From the antimicrotubular effect of colchicine it may be deduced that microtubules exert a regulative function in the formation of surface differentiations on absorptive cells of the small intestine and in the maintenance of the polarity of the cells.

Animals↗

Effect of colchicine on rat small intestinal absorptive cells. II. Distribution of label after incorporation of [3H]fucose into plasma membrane glycoproteins.

By means of radioautography the influence was tested of various periods (5, 15, 30, 40 min, 2 hr) of pretreatment with colchicine, administered intraperitoneally to rats at a dosage of 0.5 mg/100 g of body weight, on the intracellular pathway of [3H]fucose in absorptive cells of the small intestine. Administration of colchicine for 30 min and longer time intervals causes delay in the insertion of [3H]fucose into the oligosaccharide chains of glycoconjugates in the Golgi apparatus, and results in redistribution of the label apparent over the different portions of the plasma membrane. In controls, at 2 and 4 hr after administration of [3H]fucose the apical plasma membrane is strongly labeled; 53.7 +/- 3.2% of the silver grains are recorded over apical regions of the plasma membrane that contrast to basolateral portions comprising 25.4 +/- 3.2% of the label. Colchicine causes equalization of the reaction of apical and basolateral regions of the plasma membrane: the number of silver grains attributable to the apical plasma membrane is reduced; following treatment with colchicine, apical portions of the plasma membrane comprise 31.6 +/- 1.8% of the silver grains, 38.6 +/- 3.8% are attributable to basolateral membrane regions. The colchicine-induced equalization of the density of label of apical and basolateral regions of the plasma membrane, in addition to the occurrence of basolateral microvillus borders (demonstrated in the companion paper), suggests microtubules to be important in the maintenance of the polar organization of small intestinal absorptive cells.

Animals↗

Effect of colchicine on the Golgi complex of rat pancreatic acinar cells.

Colchicine administered to adult rats at a dosage of 0.5 mg/100 g of body weight effected a disorganization of the Golgi apparatus in pancreatic acinar cells. The results obtained after various periods of treatment (10 min to 6 h) showed (a) changes in all components of the Golgi complex, and (b) occurrence of large vacuoles that predominated in cytoplasmic areas outside the Golgi region. The alterations in Golgi stacks concerned elements of the proximal and distal side: (a) accumulation of transport vesicles, (b) formation of small, polymorphic secretion granules, and (c) alterations in the cytochemical localization of enzymes and reaction product after osmification. Transport vesicles accumulated and accompanied short, dilated cisternae, which lack mostly the reaction products of thiamine pyrophosphatase, inosine diphosphatase, and acid phosphatase, and osmium deposits after prolonged osmification. After 4 to 6 h of treatment, accumulated transport vesicles occupied extensive cellular areas; stacked cisternae were not demonstrable in these regions. The changes on the distal Golgi side included GERL elements: condensing vacuoles were diminished; they were substituted by small, polymorphic zymogen granules, which appeared to be formed by distal Golgi cisternae and by rigid lamellae. Unusually extended coated regions covered condensing vacuoles, rigid lamellae, and polymorphic secretion granules. A cytochemical distinction between Golgi components and GERL was possible neither in controls nor after colchicine treatment. The cytochemical alterations in Golgi components were demonstrable 20-30 min following administration of colchicine; at 45 min, initial morphological changes--augmentation of transport vesicles and formation of polymorphic zymogen granules--became apparent. 20 min after administration of colchicine, conspicuous groups of large vacuoles occurred. They were located mostly in distinct fields between cisternae of the endoplasmic reticulum, and were accompanied by small osmium--reactive vesicles. Stacked cisternae were not demonstrable in these fields. Vacuoles and vesicles were devoid of reaction products of thiamine pyrophosphatase, inosine diphosphatase, and acid phosphatase. The results provide evidence that formation of stacked Golgi cisternae is impaired after colchicine treatment. The colchicine--induced disintegration of the Golgi complex suggests a regulatory function of microtubules in the organization of the Golgi apparatus.

Acid Anhydride Hydrolases↗

The trans Golgi face in rat small intestinal absorptive cells.

In the small intestine cell differentiation from immature crypt cells to mature absorptive cells localized along the villi is accompanied by alterations in the organization of the trans Golgi side. In immature crypt cells the transmost Golgi cisterna is usually located closely adjacent to the other cisternae thus being a component of the stack. Concomitantly with cell differentiation the transmost cisterna of an increasing number of Golgi stacks sets off from the other cisternae being then located at various distances to the stacks. This transmost cisterna has, as in several other cell types, been interpreted as "GERL" (Golgi associated endoplasmic reticulum lysosomes [20, 28]) and thus, has been postulated to represent a specialized region of the endoplasmic reticulum. Our results, however, have shown that the cytochemical staining pattern which has been used as a basis for the differentiation of GERL from Golgi components is not present in crypt cells nor in mature absorptive cells of the proximal small intestine: identical cisternae react for thiamine pyrophosphatase, inosine diphosphatase, and acid phosphatase. Thiamine pyrophosphatase and inosine diphosphatase--enzymes characteristic for Golgi cisternae--are apparent over transmost cisternae defined as GERL, too, and in addition, acid phosphatase--postulated as GERL-marker--is demonstrable over stacked Golgi cisternae. This overlapping cytochemical reaction, as well as the alterations during cell differentiation, indicate that those structures which have been described as GERL are to be interpreted as Golgi components rather than as endoplasmic reticulum. On the other hand, endoplasmic reticulum is a constant component of the trans Golgi face in undifferentiated crypt-base cells and in maturing cells of the crypt-top region. From its localization closely adjacent to trans Golgi cisternae it may be termed "Golgi-associated endoplasmic reticulum"; however, these cisternae of endoplasmic reticulum are constantly devoid of acid phosphatase. No indications exist for continuities with the thiamine pyrophosphatase-, inosine diphosphatase-, and acid phosphatase-positive transmost Golgi cisternae, and for an engagement in production of lysosomes.

Acid Anhydride Hydrolases↗

Isolation of membrane-associated folded chromosomes from Anacystis nidulans.

Particles containing folded DNA were isolated from the blue-green alga Anacystis nidulans. The structure of these particles is vesicle-like and similar to that of membrane-associated nuclear bodies which had been isolated from Escherichia coli under comparable conditions. The sedimentation constant is between 8000 and 9000 Svedbergs. The DNA is inside the particles and is attached to the thylakoid membranes.

Cell Fractionation↗

The influence of the cell cycle on structure and number of nucleoli in cultured human lymphocytes.

The nucleoli of lymphocytes undergo a typical sequence of structural changes after stimulation by phytohaemagglutinin. These changes are independent of the cell cycle. Neither the inhibition of DNA-synthesis (by adenosine and methotrexate), nor the elimination of postmitotic interphase nuclei (by a colchicine block of mitoses), nor the release from such blocks has a noticeable effect on nucleolar structure or on the sequence of nucleolar changes. The number of nucleoli per cell is clearly influenced by the cell cycle. Mitosis leads to a marked increase in the number of nucleoli, whereas in all stages of interphase a decrease occurs.

Cell Nucleolus↗

The Golgi apparatus of rat small intestinal absorptive cells. I. Morphology and cytochemical staining pattern in proximal and distal small intestinal regions.

Comparative studies on the Golgi apparatus of rat small intestinal absorptive cells from duodenal, jejunal and ileal segments showed that the characteristic polarity of Golgi stacks is apparent along the entire small intestine; dilated cisternae containing lipoprotein particles are preferably localized at one Golgi side (cis face); narrow, regular cisternae predominate at the other side (trans face). Osmium deposits after prolonged osmification are restricted to cis Golgi cisternae. Pronounced differences between proximal and distal small intestinal regions exist in the cytochemical staining pattern of Golgi stacks after localization of thiamine pyrophosphatase, inosine diphosphatase and acid phosphatase. In duodenal and proximal jejunal absorptive cells demonstration of thiamine pyrophosphatase and inosine diphosphatase causes heavy deposits of reaction product over all Golgi cisternae; in the distal jejunum and in the ileum only 1-3 cisternae at the trans Golgi side are reactive. Similarly, the number of acid phosphatase-positive cisternae decreases from proximal to more distal small intestinal regions. Thiamine pyrophosphate, nucleoside diphosphatase and acid phosphatase are assumed to be active in glycosylation being involved in the removal of uridine diphosphate and other nucleoside phosphates (Tartakoff, 1980; Farquhar and Palade, 1981; Roth and Berger, 1982). Accordingly, the differences in the Golgi apparatus staining pattern after localization of these enzymes between proximal and distal small intestinal segments may reflect regional differences in the glycosylation of apoproteins in lipoprotein particles and in the kinetics of glycosylated plasma membrane and lysosomal constituents.

Acid Anhydride Hydrolases↗

The Golgi apparatus of rat small intestinal abortive cells. II. Morphology and cytochemical staining pattern during cell differentiation.

Differentiation of small intestinal absorptive cells is accompanied by morphological alterations of the Golgi apparatus which reflect the extension of its functional tasks. Golgi stacks of undifferentiated crypt cells are composed of a group of 4-7 cisternae with decreasing luminal width from one side (cis face) to the other side (trans face). In maturing cells of the upper crypt region vacuolar-like dilatations of cisternae become prominent. In mature absorptive cells dilated cisternae predominantly localized at the cis side contain lipoprotein particles thus reflecting their engagement in processing and packaging of lipoproteins. In the proximal small intestine in contrast to distal small intestinal regions, the cytochemical staining pattern of Golgi stacks after localization of thiamine pyrophosphatase, inosine diphosphatase and acid phosphatase alters coincidently with cell differentiation. In undifferentiated crypt cells only few trans Golgi cisternae exhibit reaction product. Mature absorptive cells show all cisternae strongly reactive for thiamine pyrophosphatase and inosine diphosphatase, and the majority of cisternae reactive for acid phosphatase. The alterations in the Golgi apparatus reaction pattern after localization of enzymes, assumed to be active in glycosylation (Tartakoff, 1980; Farquhar and Palade, 1981; Roth and Berger, 1982), may express alterations in the functional tasks during cell differentiation, and may reflect differences in the kinetics of glycosylated plasma membrane and lysosomal constituents between immature crypt cells and mature absorptive cells.

Acid Anhydride Hydrolases↗

Effect of colchicine on the Golgi apparatus and on GERL of rat jejunal absorptive cells. Ultrastructural localization of thiamine pyrophosphatase and acid phosphatase activity.

Ultrastructural localization of thiamine pyrophosphatase (TTP) and acid phosphatase (AcPase) activity was performed on jejunal absorptive cells of rats pretreated with the antimicrotubular agent colchicine and of control animals. Demonstration of TPP activity showed that most of the dislocated Golgi stacks after colchicine application lacked positively staining cisternae of the mature side. This cytochemical finding is in agreement with the morphologically demonstrable changes of the Golgi stacks resulting in a loss of polarity and give evidence for a colchicine-induced deficiency of the Golgi apparatus. The cytochemical localization of AcPase activity showed deposits of reaction product over lysosomes and GERL and demonstrated a dislocation of GERL occurring concomitantly with the changes of the Golgi apparatus. The antimicrotubular effect of colchicine is well documented; thus the morphological and cytochemical changes of the Golgi apparatus and of GERL might be due to a disturbed microtubular function after application of this agent suggesting an influence of microtubules in the maintenance of the integrity of these organelles. This hypothesis includes the possibility of an involvement of microtubules in formation and differentiation of Golgi stacks and GERL as well as a kind of "skeletal"function being responsible for their characteristic structure and fashion.

Acid Phosphatase↗

Nucleolar changes in human phytohaemagglutinin-stimulated lymphocytes.

The nucleoli of lymphocytes from circulating peripheral blood and from phytohaemagglutinin (PHA)-stimulated cultures (from 2 h-96h) were studied using a silver method, RNA-specific fluorescent staining, and electron microscopy of ultrathin sections. In peripheral blood about 75% of the lymphocytes have one "ring-shaped" nucleolus composed of a distinct fibrillar centre surrounded by a dense pars fibrillaris and little granular material; the remaining lymphocytes showing two or more small "ring-shaped" nucleoli. With PHA stimulation, the number of cells with several nucleoli increases first (from 2 h--12 h). Next, cells containing one or, at most, two large nucleoli with nucleolonema devoid of fibrillar centers are seen (from 4 h on). 34 h after PHA, nucleoli of the "compact" type containing one or more fibrillar centres appear and comprise about 60% of the cells after 72 h. The appearance of more than one nucleolus per cell shortly after PHA administration suggests an activation of additional nucleolar organizer regions (NOR), which fuse to form one or two large nucleoli with nucleolonema. These are then transformed into "compact" nucleoli. The fibrillar centers stasin preferentially with silver. They contain nonchromosomal proteins and may serve as stores for nucleolar proteins. The fusion of activated NORs during the first cell cycle explains the relatively high frequency of satellite associations in first mitoses compared to later mitoses after stimulation.

Cell Nucleolus↗

Effect of different preparation procedures on the ultrastructure of gonococci.

Culture samples of Neisseria gonorrhoeae and Escherichia coli were studied by transmission and electron microscopy to evaluate the effect of different preparation procedures on the pilation of these organisms. Unfixed as well as fixed bacteria showed only few, long, filamentous appendages when investigated in ultrathin sections, negatively stained specimens, or critical-point dried preparations. Snap-frozen specimens of E. coli showed many short and thin pili after being shadowed with carbon and platinum whereas those of N. gonorrhoeae showed only some type-C-like pili. Thus, the number and morphological appearance of pili appear to be greatly influenced by the preparation techniques used for study by electronmicroscopy. Conclusions as to the type and the infectivity of a bacterial strain can, therefore, not be based on purely morphological criteria.

Bacteriological Techniques↗

Endocytic routes to the Golgi apparatus.

The endocytic routes of labelled lectins as well as cationic ferritin were studied in cells with a regulated secretion, i.e. pancreatic beta cells, and in constitutively secreting cells, i.e. fibroblasts and HepG2 hepatoma cells, paying particular attention to routes into the Golgi apparatus. Considerable amounts of internalised molecules were taken up into the trans Golgi network (TGN) and into Golgi subcompartments in all three cell types as well as in secretory granules of the pancreatic beta cells. The internalised materials did not pass rapidly the TGN and Golgi stacks, but were still present hours after internalisation, being then particularly concentrated in TGN-elements and in the transmost Golgi cisterna. Endocytosed materials reached forming secretory granules present in the TGN. Further, direct fusion between endocytotic vesicles and mature secretory granules was observed. Golgi subcompartments as well as endocytic TGN containing endocytosed materials were in close apposition to specialised regions of the endoplasmic reticulum. The Golgi apparatus including its parts containing endocytosed materials were transformed into a tubular reticulum upon treatment with the fungal metabolite Brefeldin A. Rarely, internalised material was observed in the lumen of the endoplasmic reticulum, thus providing evidence for an endocytic plasma membrane to endoplasmic reticulum route.

Animals↗