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.     

Studies on the effects of vinblastine and colchicine on Trypanosoma gambiense

Vinblastine and colchicine induce the anucleate form of Trypanosoma gambiense. Light microscopic studies indicated that the anucleate form was not always produced as a result of inhibition of nuclear duplication, but was formed as a result of delay or inhibition of separation of the two nuclei after completion of nuclear division. Studies showed that vinblastine and colchicine caused disorder in arrangement of axonemal microtubules of the extracellular flagella and increased formation of both protofilaments and the axoneme composed of protofilaments in trypanosomes. Moreover, treatment with colchicine resulted in disintegration of previously existing pellicular microtubules and formation of cytoplasmic projections that appeared as protrusions from a small part of the surface membrane.     

Effect of colchicine and vinblastine on identified leech neurons

An identified neuron of the leech central nervous system is affected by the application of colchicine or vinblastine to its axon. It develops characteristic changes of membrane electrical properties, which are similar to those observed after surgical axotomy. The ionic mechanisms associated with the impulses induced by axotomy and colchicine treatment are not equivalent.     

An effect of colchicine on galactosyl- and sialyltransferases of rat liver Golgi membranes

Colchicine inhibited the activity of the galactosyl- and sialyltransferases of rat liver Golgi membranes. The sialyltransferase was more sensitive to the drug than galactosyltransferase since it was inhibited to a greater extent and at lower concentrations of colchicine than the galactosyltransferase. Two soluble enzymes, i.e. that from rat serum and that isolated from bovine milk, were not inhibited by colchicine. Even with very high concentrations of colchicine a marked stimulation of activity was observed. The data suggest that the inhibition observed in the Golgi membranes is in some way related to the arrangement of the enzymes in the lipid bilayer. In support of this hypothesis, the milk galactosyltransferase became very sensitive to colchicine after incorporation of the enzyme into lipid vesicles. The incorporation of colchicine into Golgi membranes was shown to decrease the order parameter as determined by electron spin resonance which reflects an increased fluidity of the Golgi membranes. A change in fluidity may be responsible for the inhibition of enzyme activity at least in part.     

Influence of vincristine on the Golgi complex of leukaemic lymphoblasts

In vitro and in vivo effects of vincristine on the Golgi complex of leukaemic lymphoblasts were studied. The cells incubated in vitro for 4 hours with vincristine of 1.25 x 10(-5) M concentration lacked microtubules, but regularly contained paracrystals and parallel arrays of macrotubules associated with ribosomes. The Golgi complex in control lymphoblasts was represented by 1-3 dictyosomes (stacks of cisternae) grouped in one area. After exposure to vincristine the dictyosomes lay at a considerable distance from each other. In many of them the cisternae were shorter than in controls and distended or transformed into large vacuoles. In cells incubated in vitro with lower concentrations of vincristine (1.25 x 10(-6) and 1.25 x 10(-7) M) and in cells obtained after the second therapeutic dose of vincristine (in the course of normal clinical treatment) neither changes in the Golgi complex nor formation of paracrystals and macrotubules were observed.     

Effects of vinblastine and colchicine on the rat adrenal cortex: morphometric and cytochemical studies

The effects of administration of anti-microtubular drugs--vinblastine and colchicine--on the ultrastructure of the zona fasciculata cells of young rat adrenal were studied. Young male rats were injected with vinblastine and sacrificed 2 hr later or with colchicine and sacrificed 3 hr after drug administration. Animals injected with isotonic saline in same experimental conditions served as controls. Ultrastructural alterations provoked by both drugs, vinblastine or colchicine, were identical and were most prominent in the Golgi areas. They appeared enlarged and crowded with round, or slightly elongated light vesicles, acid phosphatase, and osmium negatives. The Golgi dictyosomes, although keeping their normal morphology, were less numerous and presented cisternae which were narrower and shorter than controls. Electron-dense vesicles, round or elongated, and acid phosphatase positive--lysosomes--were observed in great number in the Golgi areas, intermingled with light vesicles. The relative volume of light vesicles and lysosomes of the treated animals was significantly increased when compared with controls, but the relative volume of dictyosomes was significantly decreased. Also the numerical density of light vesicles and lysosomes of the injected rats was significantly increased when compared with controls. These alterations are highly suggestive of the Golgi involvement in the adrenal secretory process.     

Chondroptosis: An immunohistochemical study of apoptosis and Golgi complex in chondrocytes from human osteoarthritic cartilage

The Golgi complex is thought to play an important role in the apoptotic process of osteoarthritic (OA) chondrocytes. However, the exact relationship between modifications of the Golgi complex and apoptosis in human OA cartilage requires to be established. We compared the patterns and immunolabeling intensities for anti-Golgi 58 K protein with apoptosis markers such as TUNEL and caspase-2L in OA cartilage removed from patients during knee total replacement surgery. We observed important modifications in labeling of the Golgi 58 K protein in OA chondrocytes compared with normal cell. Immunohistochemical analysis revealed co-localization between 58 K protein and caspase-2L, suggesting that this enzyme was localized in Golgi complex of OA chondrocytes. In addition, these cells labeled positive with the TUNEL technique, but in different proportions to caspase-2L. Our results support the concept, previously reported, that apoptosis in OA cartilage (chondroptosis) might be a variant of the classical apoptosis.     

Alteration of hexose transport in avian erythrocytes by vinblastine and colchicine

Vinblastine and colchicine, compounds which effect the state of aggregation of microtubules, were investigated to determine if changes in the rate of sugar transport were produced by these compounds. Vinblastine accelerated 3-O-methylglucose entry into avian erythrocytes. At a concentration of 1.5 mm, transport was accelerated two-fold. The effect of vinblastine was not attributed to cell energy depletion or to increased entry by simple diffusion. Stimulation of transport did not require preincubation of the cells with vinblastine, and the effect was reversible. Colchicine (2 mm) inhibited 3-O-methylglucose entry in aerobic or anoxic intact red cells and in red cell ghosts. A change in the state of aggregation or activity of the microtubular system could represent a model for regulation of a membrane carrier. The present results would lend support to this model.     

Morphogenesis of the hydrogenosome: An ultrastructural study

Summary— The morphogenesis of hydrogenosomes in several trichomonad species (Tritrichomonas foetus, Trichomonas vaginalis, Tritrichomonas suis, Trichomonas gallinae, Tritrichomonas augusta and Monocercomonas sp) was investigated by transmission electron microscopy of thin sections and freeze-fracture replicas of whole cells or the isolated organelle. Close proximity, and even continuity, between endoplasmic reticulum and hydrogenosomes was observed. Structures were seen connecting hydrogenosomes to each other and to cytoplasmic structures. Morphological evidence is presented showing that in all the trichomonads here studied, hydrogenosomes, like mitochondria, may divide by two distinct processes: segmentation and partition. In the segmentation process, the hydrogenosome grows, becoming enlongated with the appearance of a constriction in the central portion. Microfibrillar structures appear to help the furrowing process, ending with a total fission of the organelle. In the partition process, the division begins by an invagination of the inner hydrogenosome membrane, forming a transversal septum, separating the organelle matrix into two compartments. We suggest that myelin-like structures seen either in close contact with or in the vicinity of the hydrogenosomes may be a source of membrane lipids for hydrogenosome growth.     

Effect of colchicine and pilocarpine on the secretory activity of rat type II alveolar cells: an ultrastructural study

Colchicine in a total dose of 0.6 mg/100 g body weight per day was shown to reduce the level of apical surfactant secretion by type II alveolar cells in random-bred male albino rats, thereby demonstrating that the cytoplasmic microtubules participate in the release of surfactant into the alveolar lumen. In addition, basal secretion of surface-active material was found in 51% of all the cells. In a single dose of 8 mg/100 g b.w., pilocarpine stimulated apical surfactant secretion. If injected after colchicine, it slightly increased the number of type II alveolar cells ready to release surfactant, but actual secretion was not observed; the level of basal secretion did not increase. It has been suggested that microtubular function is not completely responsible for basal secretion and is only partly responsible for apical surfactant secretion.     

Influence of substrate curvature on osteoblast orientation and extracellular matrix deposition

Background

The effects of microchannel diameter in hydroxyapatite (HAp) substrates on osteoblast behavior were investigated in this study. Microchannels of 100, 250 and 500 μm diameter were created on hydroxyapatite disks. The changes in osteoblast precursor growth, differentiation, extra cellular matrix (ECM) secretion and cell attachment/orientation were investigated as a function of microchannel diameter.

Results

Curvature did not impact cellular differentiation, however organized cellular orientation was achieved within the 100 and 250 μm microchannels (mc) after 6 days compared to the 12 days it took for the 500mc group, while the flat substrate remained disorganized. Moreover, the 100, 250 and 500mc groups expressed a specific shift in orientation of 17.45°, 9.05°, and 22.86° respectively in 24 days. The secreted/mineralized ECM showed the 100 and 250mc groups to have higher modulus (E) and hardness (h) (E?=?42.6GPa; h?=?1.6GPa) than human bone (E?=?13.4-25.7GPa; h?=?0.47-0.74GPa), which was significantly greater than the 500mc and control groups (p?<?0.05). It was determined that substrate curvature affects the cell orientation, the time required for initial response, and the shift in orientation with time.

Conclusions

These findings demonstrate the ability of osteoblasts to organize and mineralize differentially in microchannels similar to those found in the osteons of compact bone. These investigations could lead to the development of osteon-like scaffolds to support the regeneration of organized bone.

     

An ultrastructural study of histochemical staining of complex carbohydrates in the mouse posterior vitreous body

The vitreous body contains complex carbohydrates that can be demonstrated morphologically. Vitreous hyaluronic acid is very soluble but it can be precipitated by cetylpyridinium chloride (CPC) while being cross-linked by glutaraldehyde. Oligosaccharide chains of vitreous glycoproteins are fixed with glutaraldehyde alone. Mouse eyes were fixed with glutaraldehyde or glutaraldehyde and CPC and the complex carbohydrates of the posterior vitreous cortex were studied by electron microscopy. Cationic dyes were used in the fixative or for block-staining on most fixed tissue blocks to allow detailed observations of complex carbohydrates. Most blocks were postfixed with OsO4. The hyaluronic-acid domain on vitreous collagen fibrils sequentially contracted and expanded in size with various histochemical manipulations. Contraction of the domain of hyaluronic acid generally indicates an increased charge density. OsO4 contributes considerable charge density upon forming osmate esters, but tissue postfixed with OsO4 contained large globular forms of hyaluronic acid rather than the small globules observed in non-osmicated preparations. A model is proposed to explain the seemingly paradoxical findings by reference to suggested mechanisms of polysaccharide-ligand-OsO4 interactions.     

Membrane traffic: a glitch in the Golgi matrix

Golgins are coiled-coil proteins thought to form a matrix important for shaping and organising Golgi cisternae and directing long-range recognition events in vesicular transport. This model is brought into question by new evidence that two golgins, GM130 and golgin-84, contribute to but are not essential for protein transport and Golgi structure.     

An ultrastructural study of iron and manganese deposition associated with extracellular polymers of pedomicrobium-like budding bacteria

Morphological characteristics of two Pedomicrobium-like budding bacteria are described. A structured surface layer was regularly observed on strain 868. Ruthenium red- and Alcian blue-staining polymers were found on both strains.When either strain was grown in the presence of iron or manganese, the corresponding oxides accumulated on their surfaces. In thin sections iron oxides appeared as fine threads, arrays of particles or dense coatings, depending on the source of iron. Manganese oxides appeared as branching filaments or convoluted ribbons. Both metal oxides stained with ruthenium red. Extraction of the oxides followed by ruthenium red staining revealed that polyanionic polymers previously deposited on the cells were associated with the metals.Treatment of cultures with glutaraldehyde, HgCl₂, or heat, inhibited manganese but not iron deposition, suggesting that iron oxides accumulated by passive, non-biological processes. Manganese oxides apparently accumulated under control of a biological manganese-oxidizing factor. Incomplete inhibition of manganese deposition observed in cell suspensions suggested that, if the oxidizing factor was an enzyme, it was unusually stable.Based on these results, possible mechanisms of iron and manganese deposition in association with extracellular polymers are suggested.