Translocation of Arf1 to the Secretory Granules in Rat Parotid Acinar Cells

1998 ◽  
Vol 357 (1) ◽  
pp. 147-154 ◽  
Author(s):  
Yoko Dohke ◽  
Miki Hara-Yokoyama ◽  
Junko Fujita-Yoshigaki ◽  
Richard A. Kahn ◽  
Yasunori Kanaho ◽  
...  
1984 ◽  
Vol 32 (4) ◽  
pp. 403-412 ◽  
Author(s):  
A R Hand ◽  
C Oliver

The structure and cytochemistry of the Golgi apparatus and GERL of rat parotid acinar cells was studied after in vivo secretory stimulation with isoproterenol. Discharge of mature secretory granules was complete within 1 hr after isoproterenol injection, but immature granules in the Golgi region or near the lumen were not released. At early times (1-5 hr) after isoproterenol, acid phosphatase (AcPase) activity was markedly increased in GERL and immature secretory granules compared to uninjected controls. GERL appeared increased in extent and numerous continuities with immature granules were observed. Reaccumulation of mature secretory granules was first evident at 5 hr, and was almost complete by 16 hr after isoproterenol. Thiamine pyrophosphatase (TPPase) activity, normally restricted to the trans Golgi saccules, was frequently present in immature granules during this time. Narrow cisternae resembling GERL, occasionally in continuity with immature granules, also contained TPPase reaction product. By 16-24 hr after stimulation, the activity and distribution of AcPase and TPPase were similar to control cells. These results demonstrate the dynamic nature of the Golgi apparatus and GERL in parotid acinar cells, and emphasize the close structural and functional relationship between these two structures.


2015 ◽  
Vol 60 (4) ◽  
pp. 642-649 ◽  
Author(s):  
Osamu Katsumata-Kato ◽  
Megumi Yokoyama ◽  
Miwako Matsuki-Fukushima ◽  
Takanori Narita ◽  
Hiroshi Sugiya ◽  
...  

1984 ◽  
Vol 32 (5) ◽  
pp. 477-485 ◽  
Author(s):  
A I Doine ◽  
C Oliver ◽  
A R Hand

Morphological and cytochemical changes in the Golgi apparatus and GERL of differentiating parotid acinar cells were examined in Sprague-Dawley rats from 5 days to young adult. At day 5, the Golgi apparatus consisted of 3-6 narrow saccules, with short segments of GERL lying adjacent to the trans Golgi saccule. As the glands matured, the Golgi apparatus increased in size and the saccules became broadened and fenestrated reaching a maximum from days 15-20. The saccules subsequently narrowed slightly and by day 25 resembled those seen in young adults. Numerous cisternae of GERL could be seen at the trans face during this period. While the glands were maturing, marked changes occurred in the distribution of thiamine pyrophosphatase (TPPase) activity in the Golgi saccules. In the immature cells, TPPase activity was restricted to 1 or 2 trans Golgi saccules. However, by day 10 TPPase could also be localized in immature secretory granules and in GERL-like cisternae. Unreactive segments of GERL were also present. This pattern of localization persisted until day 20, after which the TPPase activity in the GERL-like cisternae diminished gradually until by day 40 TPPase again was localized in 1-2 trans Golgi saccules and an occasional immature secretory granule. Acid phosphatase (AcPase) activity was localized primarily in lysosomes in the very young animals and increased in GERL with age up to day 15. From days 15 to 20 there was a decrease in the amount of activity seen in GERL, but from day 20 on, the AcPase activity increased until it reached that seen in young adult animals. These results indicate that the presence of TPPase activity in GERL-like cisternae and immature secretory granules may be dependent upon the developmental as well as the physiologic state of the acinar cells and lend further support to the suggestion that GERL is derived from the trans Golgi saccules.


1974 ◽  
Vol 16 (2) ◽  
pp. 309-331
Author(s):  
J. M. RADLEY

The mitotic arrest of acinar cells in the rat parotid gland in response to isoprenaline has been investigated at the ultrastructural level. The arrested cells were characterized by the presence of both centriole pairs at the cell centre, around which chromosomes formed an approximately spherical array. Microtubules radiated out from both centriole pairs, indicating that they were part of a bipolar system. The microtubule population included both non-kinetochore and kinetochore microtubules. Metaphase arrest appeared to be due to failure of the poles to separate, or to remain separated in the case of cells already in metaphase at the time of drug administration. This could be explained by the drug interfering with the ability of interpolar microtubules from opposite poles to interact with one another. The distribution of the chromosomes around the centrioles appears to depend on the presence of kinetochore microtubules, which are thought to act as tethers, either by themselves or in conjunction with non-kinetochore microtubules. It is suggested that tension is necessary between the kinetochores and poles to attain the arrest configuration. Chromosomes were observed in which both kinetochores had attached microtubules but whether or not they were linked to opposite poles remains to be investigated. Two types of arrested cell were distinguished by their content of secretory granules. Cells already in mitosis at the time of isoprenaline administration were not depleted by drug action, and during the period of arrest numerous large secretory granules were intermingled with the chromosomes. Those cells which entered mitosis after the drug was given were initially blocked in the G2 phase, during which time they were depleted of secretory granules. On entering mitosis, these cells possessed only small secretory granules, which were newly synthesized. In the metaphase-arrested state the small secretory granules were clustered around the centriolar complexes, demonstrating the presence of a poleward force operating on them. It is possible that the same force acts on the secretory granules in the first type of arrested cell, but because of the relatively large size of the granules, the clustering around the poles is not pronounced. The nature of the forces acting on the chromosomes and on the secretory granules is discussed. The functioning of these forces during the arrest state has to be considered in any general model for mitosis.


1989 ◽  
Vol 37 (11) ◽  
pp. 1645-1652 ◽  
Author(s):  
C Oliver ◽  
R Dromy ◽  
T K Hart

Exocrine acinar cells possess two cytochemically distinct populations of secondary lysosomes. One population is Golgi associated and has demonstrable acid phosphatase (AcPase) activity, whereas the second is basally located and lacks AcPase activity but has trimetaphosphatase (TMPase) activity. The basal lysosomes are tubular in shape and rapidly label with horseradish peroxidase (HRP) after intravenous injection. In the present study using isolated rat parotid acinar cells, the two lysosomal populations were separated by cell fractionation on Percoll density gradients and were analyzed biochemically and by EM cytochemistry. On 35% Percoll gradients, two peaks of AcPase and beta-hexosaminidase, both lysosomal marker enzymes, and succinic dehydrogenase, an enzyme marker for mitochondria, could be resolved. The major peaks of beta-hexosaminidase and succinic dehydrogenase and the minor peak of AcPase corresponded with the dense lysosome fraction. The major peak of AcPase and the minor peaks for beta-hexosaminidase and succinic dehydrogenase coincided with the light membrane fraction. Galactosyl transferase (a marker enzyme for Golgi saccules) and 5'-nucleotidase (a plasma membrane marker) were also associated with this fraction. By electron microscopy, the light membrane fraction was seen to contain tubular elements, multivesicular bodies (MVB), Golgi saccules, GERL, immature secretory granules, and some mitochondria. Electron microscopic cytochemical examination showed that these tubular structures were lysosomes. The dense lysosome fraction contained lysosomes positive for both AcPase and TMPase. After continuous incubation of isolated acinar cells with HRP, reaction product was rapidly localized to the light membrane fraction (greater than 2 min), where it was found in vesicles and tubular lysosomes. By 10 min it was present in MVB and tubular lysosomes, but by 60 min no HRP reaction product had appeared in the dense lysosomes. These results demonstrate that the tubular lysosomes are separable from dense lysosomes, typical secondary lysosomes, and are involved in the initial stages of endocytosis.


1984 ◽  
Vol 29 (12) ◽  
pp. 953-958 ◽  
Author(s):  
B. Carlsöö ◽  
Å. Danielsson ◽  
R. Henriksson ◽  
G. Jönsson ◽  
S. Sundström

1985 ◽  
Vol 231 (2) ◽  
pp. 431-438 ◽  
Author(s):  
S R Grant ◽  
E E Kousvelari ◽  
D K Banerjee ◽  
B J Baum

beta-Adrenergic stimulation of rat parotid acinar cells markedly increases [3H]mannose incorporation into N-linked glycoproteins [Kousvelari, Grant, Banerjee, Newby & Baum (1984) Biochem. J. 222, 17-24]. More than 90% of this protein-bound [3H]mannose was preferentially incorporated into four secretory glycoproteins. The ratio of [3H]mannose/[14C]leucine present in these individual proteins was 1.7-4-fold greater with isoproterenol-treated cells than with untreated controls. In isoproterenol-stimulated cells, [3H]mannose incorporation into mannosylphosphoryl dolichol and oligosaccharide-PP-dolichol was increased 2-3-fold over that observed in unstimulated cells. Similarly, formation of mannosylated oligosaccharide-PP-dolichol was increased approx. 4-fold in microsomes prepared from isoproterenol-treated cells. Also, turnover of oligosaccharide-PP-dolichol was significantly increased (5-fold) by β-adrenergic stimulation; the half-life for oligosaccharide-PP-dolichol decreased from 6 min in control cells to 1.2 min in isoproterenol-stimulated cells. By 15 min after isoproterenol addition to acinar cells, the specific radioactivity of parotid oligosaccharide moieties increased about 3-fold over the value observed in the absence of the agonist. Taken together, these results strongly suggest that elevation of N-linked protein glycosylation in rat parotid acinar cells after β-adrenoreceptor stimulation resulted from significant enhancement in the synthesis of mannosylphosphoryl dolichol and oligosaccharide-PP-dolichol and the turnover of oligosaccharide-PP-dolichol.


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