The pathway of intracellular transport of dehydrocholic acid was investigated in the hepatocytes of rats by transmission electron microscopy with conventional and freeze fracture methods. Both in normal and experimental groups, the cis Golgi cisterns were sacculated and faces toward the bile canaliculus. In the experimental group, however, the cis Golgi cisterns showed buds, which were probably separated to be vesicles. Some of the buds were connected to the cisterns with the narrow neck. The vesicles were increased in the vicinity of bile canaliculi. The fusion between vesicles and bile canaliculus were frequently observed in the experimental group. This was particularly well shown in the freeze fracture replica. In the thin section, the vesicles were devoid of visible contents as seen in the bile canaliculli. The evidence suggests that the vesicles are derived from the cis Gogi cistern in the way that buds pinch off, serve as vehicles to transport dehydrocholic acids and fuse to bile canaliculi for exocytosis.
Parenchyma of the cat pineal body consisted of pinealocytes and glial cells. The pinealocyte, predominant cell type, was characterized by having large mitochondria with pale matrix, abundant polyribosomes, moderately-developed Golgi apparatus, centrioles and occasional cilia. The pinealocyte had one thick and long cytoplasmic process at the one pole of the cell, and slender and shorter processes at the other pole, and in addition occasional short processes from the cell body. These processes contained longitudinally arranged microtubules, and a few mitochondria. Thick processes teminated as bulgings either in the intercellular process-rich area, or in the perivascular border which was formed by glial cell processes. These endings of pinealocyte processes had many small vesicles, mitochondria, and occasional dense bodies. Glial cells with abundant filaments of intermediate type and clear cytoplasmic matrix were fibrous astrocyte. Perikarya of the astrocytes had small and dense mitochondria, moderately developed Golgi apparatus, dense bodies and variable amount of intermediate filaments. Glial cell processes run through the intercellular spaces among the pinealocyte processes. Glial cell of protoplasmic type had no or a few filaments, but it had well-organized rough endoplasmic reticulum, dense mitochondria, well developed Golgi apparatus and many dense granules. Intercellular canaliculi formed by adjacent pinealocytes and glial cell processes were often noted. Within the parenchyma, sympathetic and parasympathetic axons and their endings were noted. These endings were present mostly in the intercellular spaces without having membrane specialization, however, in rare instances, ending with small clear and dense cored vesicles, and large dense cored vesicles formed specialized synapse with a pinealocyte process. Within the perivascular spaces nerve fibers and endings, Schwann cells and pericyte were noted. In rare case pinealocyte process penetrated into the perivascular space through the interuptions of glial border. These results suggest that pinealocyte of the cat has less significance in secretory function and is rather neural type of cell.
In this study, an attempt was made to investigate the probable organelles participating in the secretion of biligrafin. The animals (ICR male mice, 25-30gm) were divided into normal control and 6 biligrafin injected groups to which 30% biligrafin (0.006ml/gm b.w.) were injected at 10, 20, 40, 80, 160 and 320 min prior to the sampling. The mice of each group were perfused through the heart with ice-cold 2.5% glutaraldehyde buffered with 0.1M Na-cacodylate (pH. 7.4) under the Na-pentobarbital (Nembtal 0.0015mg/gm b.w.) anesthesia and liver tissues were taken from each group. Some specimens were immersed 1 hr in the same solution used in the perfusion. After an overnight rinse in 0.1M Na-cacodylate buffer containing 10% DMSO and 7.6% sucrose, $75{\mu}m$ fronzen sections were made for cytochemical study. The sections were incubated in thiamin pyrophosphatase (TPPase) and inosine diphosphatase (ID Pase) media for 70 min at $37^{\circ}C$ respectively and acid phosphatase (AcPase) medium for 40 min at $37^{\circ}C$. They were postfixed in 1 % $OsO_4$ for 1 hr. The other specimens were immersed for 8 hrs in the fixative consisting of 2.5% glutaraldehyde and 3.0% paraformaldehyde buffered with Na-cacodylate (pH. 7.4). All of the osmificated specimens were processed for electron microscopy. In both normal and biligrafin injected groups, endoplasmic reticulum (ER), vacuoles, Golgi apparatus and lysosomes were seen in the vicinity of bile canaliculus. In the biligrafin injected groups, however, the Golgi apparatus appeared to be decreased and ER and vacuoles were dilated and increased. The rough endoplasmic reticulum (RER) having a few attached ribosomes appeared to be the round saccule, especially at 20 min after biligrafin injection. Smooth endoplasmic reticulum (SER) seemed to be formed by the detachment of ribosomes at the cisternal end of RER. The cistern of SER showed saccules which probably budded off to form the vacuole. The vacuoles were devoid of visible centents. This finding seemed to be in agreement with the biochemical property of the bile constituents. The fusion between the vacuoles and bile canaliculus were frequently seen in the groups injected with biligrafin. The lysosome did not show any changes in the biligrafin injected groups. Accumulation of some material and lipid droplets were seen at the 40 and 80 min after biligrafin injection, especially at the latter. At 160 and 320 min after biligrafin injections, however, they were decreased successively while the RER stack, free ribosomes and polysomes were increased. Although the reactive products of TPPase and IDPase were observed in the ER saccules and vesicles of the normal control and biligrafin injected groups, the fusion between the bile canaliculus and saccules or vesicles could easily be seen in the latter. The AcPase activity, however, was observed in the cistern at the maturing face of Golgi apparatus and lysosomes in both normal and biligrafin groups. The results suggest that the biligrafin is excreted via the vesicles, vacuoles or sacoules probably derived from the SER without the participation of Golgi apparatus and lysosomes, and the excess amount of material is stored as inclusions during the repairing of the organelles being overactive.
Kim, Jung-Sam;Chung, Ji-Sook;Chung, Kyung-A;Roh, Young-Bok
Applied Microscopy
/
v.25
no.2
/
pp.20-28
/
1995
This study observes the change of small intestine mucosa paneth cell by changing the amount of radiation to rat. It uses the rat(Wistar) of 250-300g as the experimental animal and irradiation equipment is Gammacell 3000Elan System. and the irradiation is conducted for 500Rad group for 34sec., 1000rad for 68sec., and 1500Rad for 102sec. once on the whole body of each group, eachgroup is anesthetized with ether after 24hours. its small intestine is extrated and then it is observed by transmission electronic microscopy. The experimental results are as follows : 1. 500 Rad Group The Slightly elongated form of mitochondria and rough endoplasmic reticulum are observed in 500 Rad group. 2. 1000 Rad Group Golgi apparatus is appeared as the extended plasmodium, secretory granules exist only external membrane due to the self-fusion, the number of mitochondria that are changed as L-type are reduced, rough endoplasmic reticulum is distributed with the expanded form. 3. 1500 Rad Group The number of Golgi apparatus and granules is remarkably reduced, mitochondria is changed into C-type and free ribosomes can be observed instead of the reduction of rough endoplasmic reticulum.
The tissue reactions concerned in alveolar bone remodelling at the pressure zones of rat molar periodontium associated with the application of force (15 gm) to the maxillary first molar teeth of the albino rats were studied by the transmission electron microscopy. Osteoclasts referrable to bone resorption were observed thereafter 3 hour survival period and undermining resorption was generated thenceforth 2 day survival period. Bone resorption, reversal zone and new bone formation were simultaneously observed adjacent to the zone of undermining resorption in the 7 day survival period. Osteoclasts with well developed primary lysosome, ruffled border, clear zone, granules and Golgi apparatus were detected at the zone of the bone resorption, and dark and bright cells adjacent to the osteoclasts as well. Mononuclear cells and perpendicularly arranged collagenous fibers were observed in the reversal zone and, on the other hand, osteoblasts with well developed Golgi apparatus and rough endoplasmic reticulum were detected at the zone of bone formation.
This study was conducted to investigate stimulatory effect of epidermal growth factor (EGF) on nuclear maturation and the expression level of EGF-receptor (EGFR), GM-130 (a marker of Golgi apparatus), transport protein Sec61 subunit beta ($Sec61{\beta}$), and coatomer protein complex subunit gamma 2 (COPG2) in porcine oocytes. The cumulus-oocyte complexes were collected from follicle with 3-6 mm in diameter. They were incubated in medium with/without EGF for 22 h (IVM I) and subsequently incubated hormone-free medium with/without EGF for 22 h (IVM II). Nuclear maturation state was checked by aceto-orcein stain. Protein expression of EGFR, GM-130, $Sec61{\beta}$, and COPG2 were measured by immunofluorescence. In results, nuclear maturation of oocytes in EGF non-treated oocytes were significantly lower than EGF-treated groups at IVM I or IVM II stage (P<0.05), whereas maturational rate in EGF treatment groups at both of IVM stage was higher in among the all treatment groups (P<0.05). EGFR, GM-130, $Sec61{\beta}$ and COPG2 were expressed in the cytoplasm of oocytes. Especially, GM-130 and EGFR were strongly expressed, but $Sec61{\beta}$ and COPG2 were weakly expressed in cortical area of cytoplasm. The protein level of GM-130, $Sec61{\beta}$, and COPG2 were significantly higher in the EGF-treated groups (P<0.05). However EGFR was no difference between non EGF-treated groups and control. In conclusion, EGF plays an important role in the systems for oocyte maturation with endoplasmic reticulum and Golgi apparatus. In addition, the protein levels of $Sec61{\beta}$ and COPG2 could be changed by EGF in the porcine oocytes during maturation.
COPI vesicles are essential to the retrograde transport of proteins in the early secretory pathway. The COPI coatomer complex consists of seven subunits, termed ${\alpha}-$, ${\beta}-$, ${\beta}^{\prime}-$, ${\gamma}-$, ${\delta}-$, ${\varepsilon}-$, and ${\zeta}$-COP, in yeast and mammals. Plant genomes have homologs of these subunits, but the essentiality of their cellular functions has hampered the functional characterization of the subunit genes in plants. Here we have employed virus-induced gene silencing (VIGS) and dexamethasone (DEX)-inducible RNAi of the COPI subunit genes to study the in vivo functions of the COPI coatomer complex in plants. The ${\beta}^{\prime}-$, ${\gamma}-$, and ${\delta}$-COP subunits localized to the Golgi as GFP-fusion proteins and interacted with each other in the Golgi. Silencing of ${\beta}^{\prime}-$, ${\gamma}-$, and ${\delta}$-COP by VIGS resulted in growth arrest and acute plant death in Nicotiana benthamiana, with the affected leaf cells exhibiting morphological markers of programmed cell death. Depletion of the COPI subunits resulted in disruption of the Golgi structure and accumulation of autolysosome-like structures in earlier stages of gene silencing. In tobacco BY-2 cells, DEX-inducible RNAi of ${\beta}^{\prime}$-COP caused aberrant cell plate formation during cytokinesis. Collectively, these results suggest that COPI vesicles are essential to plant growth and survival by maintaining the Golgi apparatus and modulating cell plate formation.
Brefeldin A (BFA), a lactone antibiotic isolated from the fungus Eupenicillium brefeldianum, inhibits the transport of secreted and membrane proteins from the endoplasmic reticulum (ER) to the Golgi apparatus. BFA disrupts Golgi function, the accumulation of unfolded proteins in ER, and the induction of ER stress. Prolonged ER stress induces apoptosis at least in part through the transcription factor C/EBP (CCAAT/enhancer binding protein) homologous protein (CHOP),which is activated by the unfolded protein response (UPR). In this paper, we demonstrate that BFA-induced endoplasmic reticulum stress leads to different CHOP expression in primary astrocyte cells and C6 glioma cells. BFA induced lower CHOP expression levels in primary astrocyte cells than in C6 glioma cells; however, other ER stress inducers (thapsigargin and tunicamycin) resulted in similar expression patterns in these two cell types. Interestingly, the three different ER stress inducers (BFA, thapsigargin, and tunicamycin) induced similar levels of CHOP mRNA expression in primary astrocyte cells. The ubiquitin-proteasome inhibitor MG132 also markedly up-regulated the BFA-mediated CHOP protein expression in primary astrocyte cells. BFA also induced higher proteasome activity in primary astrocyte cells than in C6 glioma cells. Taken together, our results suggest that higher proteasomal activity might down-regulate BFA-induced CHOP expression in primary astrocyte cells.
In order to define the morphological changes of the secreting cells of prothoracic gland during larva-pupal molt, ultrastructural observations were carried out using Bombyx mori L. as the experimental material. At first stage of present experiment, 4 day old 5th instar larva, the polyhedral secreting cells were centrally located in the prothoracic gland surrounded by the connective sheath. The secreting cells were attached to the neighboring cells by the prominent desmosomes, and the plasma membrane contacted with connective sheath were highly infolded. In cytoplasm, the most of the cell organelles, such as rod-like mitochondria, rough surfaced endoplasmic reticulum, ribosome were developed. As the stages advance from larva to pupa, general feature of the secreting cells were retained, but structural changes of the various cytoplasmic organelles-ribosome, rough surfaced endoplasmic reticulum, mitochondria, Golgi apparatus, lamellar body, and vesicle-were noted. In the perinuclear cytoplasm of the secreting cells at the stage of 6 day old 5th instar larva, it is peculiar that only a large amount of ribosomes were distributed and the other organelles were retreated from the juxtanuclear region. Just before and after spining cocoon, these features were more remarkable. Rough surfaced endoplasmic reticulum were gradually increased from the stage just before spining cocoon to the pharate pupa. Rod-like mitochondria with irregular cristae and the matrix showing low density were distributed throughout the cytoplasm in the secreting cells of the 4 day old 5th instar larva. Sometimes, longitudinally distended and curved mitochondria were observed. At the stage of pharate pupa, most of mitochondria were deformed. The rod-like mitochondria of the secreting cells of pupal prothoracic gland were narrower than those of 4 day old 5th instar larva, and the electron density of the mitochondrial matrix is increased in pupa. Golgi apparatus were a few in number in both stages, last instar larva and spining cocoon. In stages of the pharate pupa, the Golgi apparatus were frequently observed. Cytoplasmic vesicles were observed for the first time in the secreting cells of one day after spining cocoon, and the number and the size of cytoplasmic vesicles were distinctly increased inpharate pupa and just after pupation. In the secretory cells of the PG, it in concluded that the RER was closely related to syntheting the enzymes seem to produce the ecdysone.
Proceedings of the Korean Society of Applied Pharmacology
/
1998.11a
/
pp.167-168
/
1998
Swainsonine is a toxic indolizidine alkaloid found in the plant, Swainsona species (Dorling, 1978; Colgate, 1979). It is a potent inhibitor of the glycoprotein processing pathway in the Golgi apparatus. Specifically, it inhibits mannosicase II resulting in abherrant high mannose glycans. Recent studies showed that swainsonine prevents metastasis of tumor cells and it inhibits solid growth tumor, at least partially (Goss et al., 1994; Baptista et al, 1994).
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