scholarly journals Role of microtubules in the organization and localization of the Golgi apparatus.

1984 ◽  
Vol 99 (1) ◽  
pp. 113s-118s ◽  
Author(s):  
I V Sandoval ◽  
J S Bonifacino ◽  
R D Klausner ◽  
M Henkart ◽  
J Wehland
Keyword(s):  
Golgi Apparatus ◽  
Close Association ◽  
A549 Cells ◽  
Cell Periphery ◽  
Microtubule Depolymerization ◽  
Microtubule Organizing Center ◽  
Normal Medium ◽  
Large Numbers ◽  
Alpha Beta ◽  
Tubulin Paracrystals

Normal interphase PtK2 and A549 cells display long microtubules radiating from the microtubule-organizing center (MTOC) to the plasma membrane. Both MTOC and Golgi apparatus are contained in the same perinuclear area. Treatment of cells with 1 microM colcemid for 2 h results in microtubule depolymerization and fragmentation of the Golgi apparatus into elements scattered throughout the cytoplasm. Both normal microtubules and the Golgi apparatus assemble again following removal of colcemid. Injection of the alpha, beta-nonhydrolyzable GTP analog, guanosine 5'(alpha, beta-methylene)diphosphate [pp(CH2)pG], into interphase cells growing in normal medium results in the formation of microtubule bundles resistant to colcemid and prevents the fragmentation of the Golgi apparatus. Injection of pp(CH2)pG into cells incubated with colcemid results in substitution of tubulin ribbons for microtubules and has no effect on the Golgi-derived elements scattered throughout the cytoplasm. Removal of colcemid 1 h after the injection of pp(CH2)pG results in polymerization of large numbers of short, single randomly oriented microtubules, whereas the Golgi apparatus remains fragmented. Treatment of cells with 10 microM taxol for 3 h results both in polymerization of microtubule bundles without relation to the MTOC in the cell periphery and fragmentation of the Golgi apparatus. The Golgi-derived fragments are present exclusively in regions of the peripheral cytoplasm enriched in microtubules. The codistribution of microtubules and Golgi elements can be reversed in taxol-treated cells by injection of a monoclonal (YL 1/2) antibody reacting specifically with the tyrosylated form of alpha-tubulin. Cells incubated with colcemid after treatment with taxol have large numbers of Golgi-derived elements in close association with colcemid-resistant microtubule bundles. Incubation of cells with 50 microM vinblastine for 90 min results in microtubule dissembly, formation of tubulin paracrystals, and fragmentation of the Golgi apparatus into elements without relation to the tubulin paracrystals.

scholarly journals The Golgi apparatus remains associated with microtubule organizing centers during myogenesis.

1985 ◽  
Vol 101 (2) ◽  
pp. 630-638 ◽  
Author(s):  
A M Tassin ◽  
M Paintrand ◽  
E G Berger ◽  
M Bornens
Keyword(s):  
Golgi Apparatus ◽  
Nuclear Periphery ◽  
Spatial Relationship ◽  
Cell Types ◽  
Microtubule Depolymerization ◽  
Animal Cells ◽  
Microtubule Organizing Center ◽  
Microtubule Disruption ◽  
Tight Association

In vitro myogenesis involves a dramatic reorganization of the microtubular network, characterized principally by the relocalization of microtubule nucleating sites at the surface of the nuclei in myotubes, in marked contrast with the classical pericentriolar localization observed in myoblasts (Tassin, A. M., B. Maro, and M. Bornens, 1985, J. Cell Biol., 100:35-46). Since a spatial relationship between the Golgi apparatus and the centrosome is observed in most animal cells, we have decided to follow the fate of the Golgi apparatus during myogenesis by an immunocytochemical approach, using wheat germ agglutinin and an affinity-purified anti-galactosyltransferase. We show that Golgi apparatus in myotubes displays a perinuclear distribution which is strikingly different from the polarized juxtanuclear organization observed in myoblasts. As a result, the Golgi apparatus in myotubes is situated close to the microtubule organizing center (MTOC), the cis-side being situated at a fixed distance from the nuclear envelope, a situation which suggests the existence of a structural association between the Golgi apparatus and the nuclear periphery. This is supported by experiments of microtubule depolymerization by nocodazole, in which a minimal effect was observed on Golgi apparatus localization in myotubes in contrast with the dramatic scattering observed in myoblasts. In both cell types, electron microscopy reveals that microtubule disruption generates individual dictyosomes; this suggests that the connecting structures between dictyosomes are principally affected. This structural dependency of the Golgi apparatus upon microtubules is not apparently accompanied by a reverse dependency of MTOC structure or function upon Golgi apparatus activity. Golgi apparatus modification by monensin, as effective in myotubes as in myoblasts, is without apparent effect on MTOC localization or activity and on microtubule stability. The main result of our study is to show that in a cell type where the MTOC is dissociated from centrioles and where antero-posterior polarity has disappeared, the association between the Golgi apparatus and the MTOC is maintained. The significance of such a tight association is discussed.

scholarly journals Associations of elements of the Golgi apparatus with microtubules.

1984 ◽  
Vol 99 (3) ◽  
pp. 1092-1100 ◽  
Author(s):  
A A Rogalski ◽  
S J Singer
Keyword(s):  
Golgi Apparatus ◽  
Cultured Cells ◽  
Temperature Shift ◽  
Temperature Sensitive ◽  
Cell Periphery ◽  
Microtubule Organizing Center ◽  
Organizing Center ◽  
Free Microtubules ◽  
Vesicular Stomatitis ◽  
In Cells

The intracellular spatial relationships between elements of the Golgi apparatus (GA) and microtubules in interphase cells have been explored by double immunofluorescence microscopy. By using cultured cells infected with the temperature-sensitive Orsay-45 mutant of vesicular stomatitis virus and a temperature shift-down protocol, we visualized functional elements of the GA by immunolabeling of the G protein of the virus that was arrested in the GA during its intracellular passage to the plasma membrane 13 min after the temperature shift-down. Complete disassembly of the cytoplasmic microtubules by nocodazole at the nonpermissive temperature before the temperature shift led to the dispersal of the GA elements, from their normal compact perinuclear configuration close to the microtubule-organizing center (MTOC) into the cell periphery. Washout of the nocodazole that led to the reassembly of the microtubules from the MTOC also led to the recompaction of the GA elements to their normal configuration. During this recompaction process, GA elements were seen in close lateral apposition to microtubules. In cells treated with nocodazole followed by taxol, an MTOC developed, but most of the microtubules were free of the MTOC and were assembled into bundles in the cell periphery. Under these circumstances, the GA elements that had been dispersed into the cell periphery by the nocodazole treatment remained dispersed despite the presence of an MTOC. In cells treated directly with taxol, free microtubules were seen in the cytoplasm in widely different, bundled configurations from one cell to another, but, in each case, elements of the GA appeared to be associated with one of the two end regions of the microtubule bundles, and to be uncorrelated with the locations of the vimentin intermediate filaments in these cells. These results are interpreted to suggest two types of associations of elements of the GA with microtubules: one lateral, and the other (more stable) end-on. The end-on association is suggested to involve the minus-end regions of microtubules, and it is proposed that this accounts for the GA-MTOC association in normal cells.

Microtubule depolymerization inhibits transport of cathepsin D from the Golgi apparatus to lysosomes

1990 ◽  
Vol 96 (4) ◽  
pp. 711-720
Author(s):  
J. Scheel ◽  
R. Matteoni ◽  
T. Ludwig ◽  
B. Hoflack ◽  
T.E. Kreis
Keyword(s):  
Golgi Apparatus ◽  
Cathepsin D ◽  
Lysosomal Enzymes ◽  
Spatial Organization ◽  
Microtubule Depolymerization ◽  
Microtubule Organizing Center ◽  
Microtubule Network ◽  
Organizing Center ◽  
Efficient Transport ◽  
Golgi Network

Lysosomes as well as a prelysosomal compartment rich in the mannose 6-phosphate receptor are clustered close to the Golgi apparatus in the perinuclear region of the microtubule organizing center in interphase human skin fibroblasts. The spatial organization of these organelles depends on an intact microtubule network. Depolymerization of the microtubules by treatment of cells with nocodazole leads to random scattering of Golgi elements, the prelysosomal compartment, and lysosomes throughout the cytoplasm. To test whether microtubules and the spatial organization of these organelles are important for efficient transport of lysosomal enzymes, the effect of microtubule depolymerization on the maturation of newly synthesized cathepsin D was studied. An up to fivefold inhibition of proteolytic maturation of cathepsin D was observed in drug-treated cells. This effect was due to a decreased rate of transport of cathepsin D from the Golgi apparatus to lysosomes. Depolymerization of microtubules did not inhibit transport of cathepsin D from the endoplasmic reticulum to the trans-Golgi network. Furthermore, synthesis of the phosphomannosyl marker present on cathepsin D was not affected by nocodazole. These results suggest that efficient transport of cathepsin D from the Golgi apparatus to a prelysosomal compartment and lysosomes is facilitated by microtubules and the spatial organization of these organelles.

scholarly journals AP-4 vesicles contribute to spatial control of autophagy via RUSC-dependent peripheral delivery of ATG9A

2017 ◽  
Author(s):  
Alexandra K. Davies ◽  
Daniel N. Itzhak ◽  
James R. Edgar ◽  
Tara L. Archuleta ◽  
Jennifer Hirst ◽  
...  
Keyword(s):  
Membrane Trafficking ◽  
Close Association ◽  
Adaptor Protein ◽  
Cell Types ◽  
Subcellular Localisation ◽  
Cell Periphery ◽  
Accessory Proteins ◽  
Unknown Aetiology ◽  
Spatial Control ◽  
Cargo Proteins

AbstractAdaptor protein 4 (AP-4) is an ancient membrane trafficking complex, whose function has largely remained elusive. In humans, AP-4 deficiency causes a severe neurological disorder of unknown aetiology. We apply unbiased proteomic methods, including ‘Dynamic Organellar Maps’, to find proteins whose subcellular localisation depends on AP-4. We identify three transmembrane cargo proteins, ATG9A, SERINC1 and SERINC3, and two AP-4 accessory proteins, RUSC1 and RUSC2. We demonstrate that AP-4 deficiency causes missorting of ATG9A in diverse cell types, including patient-derived cells, as well as dysregulation of autophagy. RUSC2 facilitates the transport of AP-4-derived, ATG9A-positive vesicles from the TGN to the cell periphery. These vesicles cluster in close association with autophagosomes, suggesting they are the “ATG9A reservoir” required for autophagosome biogenesis. Our study uncovers ATG9A trafficking as a ubiquitous function of the AP-4 pathway. Furthermore, it provides a potential molecular pathomechanism of AP-4 deficiency, through dysregulated spatial control of autophagy.

Golgi Fragmentation in Human Patients with Chronic Atrial Fibrillation: A New Aspect of Remodeling

2018 ◽  
Vol 67 (02) ◽  
pp. 098-106 ◽  
Author(s):  
Luisa Jungk ◽  
Heike Franke ◽  
Aida Salameh ◽  
Stefan Dhein
Keyword(s):  
Atrial Fibrillation ◽  
Golgi Apparatus ◽  
Fragment Size ◽  
Chronic Atrial Fibrillation ◽  
Ionic Channels ◽  
Longitudinal Axis ◽  
Cyclin Dependent Kinase ◽  
Microtubule Depolymerization ◽  
Tissue Samples ◽  
Golgi Fragmentation

Background Atrial fibrillation (AF) is the most common chronic arrhythmia in elderly people and is accompanied by remodeling processes. While much is known about changes in ionic channels and in extracellular matrix, less is known about possible changes of intracellular structures. Objective We wanted to investigate, whether AF may also affect the structure of the Golgi apparatus and the microtubular network. Methods One-hundred fifty-three cardiac surgery patients were investigated [n = 24 in sinus rhythm (SR) and n = 129 with chronic AF of >1 year duration]. Tissue samples of the left atrial free wall were examined immunohistochemically. Golgi apparatus was detected by GM130 and its phosphorylated isoform. Furthermore, we investigated the length of the microtubules by α-tubulin staining. We also measured stathmin (phospho-S37), which is known to induce microtubule depolymerization. In addition, we investigated the cyclin-dependent kinase cdk5-activation, a typical stimulus for Golgi fragmentation, by measuring membrane-associated cdk5. Results We found significant fragmentation of the Golgi apparatus in AF together with a reduced fragment size. Significant more fragments of the Golgi were found lateral to the nucleus in AF, while the Golgi in SR was located more to the polar side of the nucleus, that is, in the longitudinal axis of the cell. This was accompanied by a significant reduction of the number of tubulin strands longer than 10 µm. These changes did not go along with an activation of stathmin, but with an increase in membrane association of cdk5. Conclusions The present data may show that AF associated remodeling also involves intracellular remodeling of the Golgi-microtubular apparatus.

Effect of shear stress upon localization of the Golgi apparatus and microtubule organizing center in isolated cultured endothelial cells

1993 ◽  
Vol 104 (4) ◽  
pp. 1145-1153 ◽  
Author(s):  
D.E. Coan ◽  
A.R. Wechezak ◽  
R.F. Viggers ◽  
L.R. Sauvage
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Cell Migration ◽  
Golgi Apparatus ◽  
Flow Direction ◽  
Microtubule Organizing Center ◽  
Directed Cell Migration ◽  
Time Period ◽  
Significant Difference ◽  
Organizing Center

Despite substantial evidence to suggest that directed cell migration is dependent upon positioning of the Golgi apparatus (GA) and the microtubule organizing center (MTOC), some controversy exists about whether such a relationship is relevant to endothelial cells under flow. The present study was undertaken to provide an indepth investigation of the relationship between shear stress, GA/MTOC localization, cell migration and nuclear position. Bovine carotid endothelial cells were exposed to 22 or 88 dynes/cm2 for 0.5, 2, 8 or 24 h, and localization of their GA/MTOCs was determined relative to the direction of flow. In no-flow control specimens, (0, 0.5, 2, 8 and 24 h) there was no change in the equally distributed GA/MTOCs. In contrast, during the first 8 h at 88 dynes/cm2 and by 2 h at 22 dynes/cm2 there was a significant increase in the number of cells with GA/MTOCs localized upstream to flow direction. The effect was temporary, however, and by 24 h there was no significant difference between the no-flow, 22 and 88 dynes/cm2 specimens. Analysis of GA/MTOC localization with respect to the direction of cell migration determined that 72.5% of no-flow cells possessed GA/MTOCs localized to the sides of nuclei nearest the direction of migration. In contrast, 64% of the specimens shear stressed over the same time period had GA/MTOCs localized to the sides of nuclei opposite the direction of migration. These results suggest that positioning of the GA/MTOC in endothelial cells is not dependent completely upon the direction of migration.(ABSTRACT TRUNCATED AT 250 WORDS)

Memoirs: A Study of the Cytoplasmic Inclusions and Nucleolar Phenomena during the Oogenesis of the Mouse

1933 ◽  
Vol s2-75 (300) ◽  
pp. 697-721
Author(s):  
R.A. R. GRESSON
Keyword(s):  
Golgi Apparatus ◽  
Close Association ◽  
Large Body ◽  
Follicle Cell ◽  
Nuclear Body ◽  
Follicle Cells ◽  
Present Contribution ◽  
Cytoplasmic Inclusions ◽  
Golgi Bodies ◽  
Nucleolar Material

1. The Golgi apparatus of the germinal epithelium consists of a dark mass of material situated at one pole of the nucleus. The mitochondria occur scattered throughout the cytoplasm. 2. The Golgi material of the very early oocyte consists of rods and granules clumped together to form a large body at one pole of the nucleus; smaller masses of Golgi material may also be present. 3. In the young oocyte, surrounded by a follicle wall, a single juxta-nuclear body is present; at a later stage the individual Golgi elements break away from the juxta-nuclear body and become distributed throughout the ooplasm. 4. In the late oocytes the Golgi elements occur in close association with the mitochondrial clumps and also scattered through the ooplasm. In tubal eggs the Golgi bodies are smaller in size and more numerous than in the ovarian ova. 5. It is concluded that the large mitochondria of Lams and Doorme correspond to the oocyte Golgi elements of the present contribution. The behaviour of the Golgi material during the growth of the ovum resembles that of the eggs of other mammals. The present findings on the structure of the juxta-nuclear Golgi material agrees with Nihoul's account for the rabbit. 6. The mitochondria of the young oocytes occur scattered through the ooplasm, but are more numerous in the vicinity of the nucleus and Golgi material. Later, the majority of the mitochondria become collected into clumps; in the tubal eggs the mitochondrial clumps are more numerous. 7. The Golgi apparatus of young follicles is situated between the follicle-cell nucleus and the pole of the cell directed towards the oocyte; in follicles consisting of several layers the position of the Golgi apparatus varies, while in fully-formed follicles the Golgi material of many of the cells surrounding the follicular cavity are directed towards the cavity. This agrees with Henneguy's findings for the Golgi apparatus of the follicle-cells of the guinea-pig. The mitochondria of the follicle-cells occur scattered through the cytoplasm but are more numerous towards the pole of the cell adjoining the oocyte. 8. The number of nucleoli present in the early oocyte varies from one to five; the majority of the older oocytes contain a single nucleolus but two may be present. Extrusion into the ooplasm of nucleolar material takes place; the nucleoli and the nucleolar extrusions are basophil (Mann's methyl-blue eosin). 9. Fatty yolk is not present in the mouse ovum. It is suggested that the Golgi elements and mitochondria play some part in yolk-formation, and that some of the granules formed by the fragmentation of the nucleolar extrusions are added to the yolkglobules already present. The yolk-globules of unsegmented tubal eggs are situated towards one pole of the cell; at the twocell stage they appear to be evenly distributed between the two cells. 10. In degenerating eggs the mitochondria are clumped; the Golgi bodies occur in small groups or are closely applied to the mitochondrial clumps. In eggs which have undergone fragmentation the Golgi bodies occur in groups, while the majority of the mitochondria are clumped. The fat-globules, previously recorded by Kingery in degenerating eggs, were identified. In material treated by Ciaccio's method for the identification of fats, appearances suggest that the Golgi elements, and possibly the mitochondria, give rise to fat. Yolk-globules could not be distinguished in the late stages of these eggs.

Cooperative defence and other benefits among exposed spiny lobsters: inferences from group size and behaviour

2001 ◽  
Vol 52 (8) ◽  
pp. 1113 ◽  
Author(s):  
William F. Herrnkind ◽  
Michael J. Childress ◽  
Kari L. Lavalli
Keyword(s):  
Drag Reduction ◽  
Group Size ◽  
Field Data ◽  
Close Association ◽  
Size Distributions ◽  
Frequency Distributions ◽  
Large Numbers ◽  
Spiny Lobsters ◽  
Migratory Period ◽  
Previous Group

Caribbean spiny lobsters show strikingly coordinated queuing behaviour and resting, outward-facing radial formations, especially during mass migrations when large numbers cross shelter-poor substrate in daylight. The close association of individual lobsters during these behaviours could be due to chance or some benefit of association such as dilution (and associated selfish-herd effects), group vigilance, cooperative defence, and/or drag reduction during migration. To infer probable beneficial functions, we examined the frequency distributions of individuals and groups using a seven-year set of field data and additional behavioural observations in large seawater enclosures. Group size distributions were not significantly aggregated in dens during the non-migratory period but became highly aggregated during migration. The group size distributions of lobsters initially leaving dens and those observed moving in the open were statistically different from one another, indicating that group sizes at each of these steps in the migration are not simply the result of previous group sizes. The distribution of group sizes suggests that, during movement in the open, dilution, vigilance, cooperative defence, and/or drag reduction may all favour the formation of queues. During resting in the open, dilution, vigilance, and cooperative defence may continue to favour individuals that remain in formation within the group.

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