• International Journal of Oral Biology
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• v.33 no.4
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• pp.125-129
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• 2008

Inositol 1,4,5-trisphosphate ($IP_3$) plays an important role in the release of $Ca^{2+}$ from intracellular stores into the cytoplasm in a variety of cell types. $IP_3$ translocation dynamics have been studied in response to many types of cell signals. However, the dynamics of cytosolic $IP_3$ in salivary acinar cells are unclear. A green fluorescent protein (GFP)-tagged pleckstrin homology domain (PHD) was constructed and introduced into a phospholipase C ${\delta}1$ (PLC ${\delta}1$) transgenic mouse, and then the salivary acinar cells were isolated. GFP-PHD was heterogeneously localized at the plasma membrane and intracellular organelles in submandibular gland and parotid gland cells. Application of trypsin, a G protein-coupled receptor activator, to the two types of cells caused an increase in GFP fluorescence in the cell cytoplasm. The observed time course of trypsin-evoked $IP_3$ movement in acinar cells was independent of cell polarity, and the fluorescent label showed an immediate increase throughout the cells. These results suggest that GFP-PHD in many tissues of transgenic mice, including non-cultured primary cells, can be used as a model for examination of $IP_3$ intracellular dynamics.

• Journal of Korean Academy of Oral and Maxillofacial Radiology
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• v.15 no.1
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• pp.27-40
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• 1985

This study was undertaken to observe the histopathologic changes in salivary gland of the white rats when exposed to megavoltage fractionated dose of cobalt-60 irradiation and 78 female white rats, weighing approximately 180gm, were divided into control and 3 experimental groups. Irradiation on experimental groups was delivered by using 6000 curies MeV ALCYON cobalt-60 teletherapy unit with exposure rate 183 rads per minute, in source skin distance 80cm, 600 rads every 3 days. In experimental groups, Group Ⅰwas irradiated of total dose 1200 rads for a period of 6 days, Group Ⅱ was irradiated of total dose 2400 rads for a period of 12 days and Group Ⅲ was irradiated of total dose of 4800 rads for a period of 24 days. The animals were sacrificed serially at 3 hours, 6 hours, 10 hours, 1st day, 4th day, 7th day after each completion of irradiation exposure. At sacrifice, salivary glands were excised and examined microscopically and electromicroscopically. The results were as follows: 1. The acinar cells of parotid and submaxillary gland showed damage varied with dose, 1200 rads resulted in very mild injury while 4800 rads caused most extensive injury. 2. The acinar cells of parotid and submandibular gland showed similar ultrastructural alterations, appeared as pleomorphic nucleus, decreased numbers and pleomorphism of secretory granules, distention of rough endplasmic reticulum, expansion and pallor appearance of mitochondria, and hypertrophy of Golgi complex. 3. Parotid serous cells were the most sensitive components, displaying morphological alterations of radiation damage as early as 3 hours, followed by submandibular seromucinous cells and secretory tubular cells. 4. The mucous cells of sublingual gland, as well as the whole ductal lining cells of each salivary gland, displayed no significant alterations. No evidence of microvascular injury through whole experimental groups indicated that microvascular impairment does not contribute to early salivary gland injury.

• Imaging Science in Dentistry
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• v.35 no.3
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• pp.147-156
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• 2005

Purpose : To observe the histopathological changes and caspase-3 expression in the submandibular gland in streptozotocin-induced diabetic rats after irradiation. Materials and Methods : The male Sprague-Dawley rats weighing approximately 250 gm were divided into four groups: control, diabetes, irradiation, and diabetes-irradiation groups. Diabetes mellitus was induced in the rats by injecting streptozotocin. Rats in the control and irradiation groups were injected with citrate buffer only. After 5days, rats in irradiation and diabetes-irradiation groups were irradiated with a single absorbed dose of 10 Gy to the head and neck region. All the rats were sacrificed at 3, 7, 14, 21, and 28 days after irradiation. The specimen including the submandibular gland were sectioned and observed using histopathological and immunohistochemical methods. Results : In the irradiation group, the condensed nucleus, karyolysis, and degeneration of the acinar cells and atrophy of the duct cells were observed in the early experimental phase. However, the acinar cells were found to be normal at 28 days after irradiation. In the diabetes group, the condensed nucleus, karyolysis, atrophy, and degeneration of the acinar cells were observed in the early experimental phase. However, the acinar cells were found to be normal at 21 days after diabetic state induction. In the diabetes-irradiation group, the ductal epithelial cells were predominant in their glandular tissues at 28 days after irradiation. In all of the experimental groups, the most prominent change of the acinar cells and ductal cells were observed at 14 days after diabetic state induction and irradiation. Conclusion The expression of caspase-3 in the acinar cells and ductal cells of the submandibular gland was weak after irradiation, but that in the acinar cells, ductal cells, and fibrous cells of the submandibular gland was prominent after diabetic state induction.

• Journal of dental hygiene science
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• v.22 no.2
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• pp.83-89
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• 2022

Background: Estrogen deficiency affects the structure and function of the salivary glands in women, leading to a decrease in salivary secretion and a change in the composition of saliva. Previous studies on changes in the salivary glands that cause estrogen deficiency have reported only partial results for the parotid and submandibular glands, and there are few comparative morphological studies of histological changes between the parotid and submandibular glands in ovariectomized rats (OVX) leading to estrogen deficiency. This study aimed to analyze the histopathological and histochemical changes in the parotid and submandibular salivary glands causing estrogen deficiency by using OVX, and to discuss the mechanism on these changes. Methods: The parotid and submandibular glands from sacrificed control and OVX groups were fixed with cold 4% paraformaldehyde in phosphate buffer (pH 7.2). The tissues were dehydrated using a series of graded ethyl alcohol and embedded in paraffin. For histopathological analysis, sections cut to a thickness of 6 to 7 ㎛ were stained with hematoxylin and eosin (H&E). For histochemical analysis, Periodic acid-Schiff (PAS), Alcian blue (AB, pH 2.5), and PAS+AB (pH 2.5 and pH 1) staining was performed. Results: Histopathological analysis of OVX tissue showed that the parotid and submandibular salivary glands were broadly and clearly separated and divided into lobes. In OVX, acinar and ductal cells with condensed polymorphic or pyknotic nucleus, which are presumed to be characteristic of apoptotic cells, and degenerated cells with lipid deposition in cytoplasmic granules and ruptured membranes were increased. Histochemical analysis of OVX, confirmed an increase in the number and acidification of acinar secretory granules. Conclusion: Histopathological and histochemical changes and the effects of estrogen deficiency are more evident in the submandibular salivary gland than in the parotid gland.

• The Journal of the Korean dental association
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• v.38 no.4 s.371
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• pp.378-384
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• 2000

• Biomedical Science Letters
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• v.10 no.3
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• pp.211-217
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• 2004

For discrimination of ductal and ascinar cells, we isolated a single-chain variable domain fragment (scFv) antibody against ductal cells of salivary gland using phage display technique. From the spleen of a mouse immunized with ductal cell lysate, total RNA was prepared and used as a template for cDNA synthesis of antibody genes. The scFv genes were constructed with variable domain genes of heavy and light chain and were introduced into pCANTAB5E to construct phage scFv library. The phage particles specific for acinar cells were screened by subtraction using immunotubes coated with acinar and ductal cell lysate and enzyme-linked immunoabsorbance assay (ELISA). The characteristics of the scFv were determined by immunohistochemistry (IHC) and the result indicated that the isolated scFv has the specificity against ductal cells of salivary glands and tubules of kidney. And the scFv has an unique binding activity specific for Hashimoto's thyroiditis. The nucleotide sequence of isolated scFv gene was determined and revealed that V/sub H/ belongs to the mouse H-chain family subgroup IB and V/sub L/ to the mouse L-chain family subgroup III.

• The Korean Journal of Physiology and Pharmacology
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• v.22 no.2
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• pp.215-223
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• 2018

Intracellular $Ca^{2+}$ mobilization is closely linked with the initiation of salivary secretion in parotid acinar cells. Reactive oxygen species (ROS) are known to be related to a variety of oxidative stress-induced cellular disorders and believed to be involved in salivary impairments. In this study, we investigated the underlying mechanism of hydrogen peroxide ($H_2O_2$) on cytosolic $Ca^{2+}$ accumulation in mouse parotid acinar cells. Intracellular $Ca^{2+}$ levels were slowly elevated when $1mM\;H_2O_2$ was perfused in the presence of normal extracellular $Ca^{2+}$. In a $Ca^{2+}-free$ medium, $1mM\;H_2O_2$ still enhanced the intracellular $Ca^{2+}$ level. $Ca^{2+}$ entry tested using manganese quenching technique was not affected by perfusion of $1mM\;H_2O_2$. On the other hand, $10mM\;H_2O_2$ induced more rapid $Ca^{2+}$ accumulation and facilitated $Ca^{2+}$ entry from extracellular fluid. $Ca^{2+}$ refill into intracellular $Ca^{2+}$ store and inositol 1,4,5-trisphosphate ($1{\mu}M$)-induced $Ca^{2+}$ release from $Ca^{2+}$ store was not affected by $1mM\;H_2O_2$ in permeabilized cells. $Ca^{2+}$ efflux through plasma membrane $Ca^{2+}-ATPase$ (PMCA) was markedly blocked by $1mM\;H_2O_2$ in thapsigargin-treated intact acinar cells. Antioxidants, either catalase or dithiothreitol, completely protected $H_2O_2-induced$ $Ca^{2+}$ accumulation through PMCA inactivation. From the above results, we suggest that excessive production of $H_2O_2$ under pathological conditions may lead to cytosolic $Ca^{2+}$ accumulation and that the primary mechanism of $H_2O_2-induced$ $Ca^{2+}$ accumulation is likely to inhibit $Ca^{2+}$ efflux through PMCA rather than mobilize $Ca^{2+}$ ions from extracellular medium or intracellular stores in mouse parotid acinar cells.

• International Journal of Oral Biology
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• v.33 no.4
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• pp.143-147
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• 2008

The sodium bicarbonate cotransporter (NBC) protein is functionally expressed in salivary glands. In this experiment, we examined the role of NBC in $HCO_3^-$ formation in human parotid gland acinar cells. Intracellular pH (pHi) was measured in 2'-7'-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF)-loaded cells. Acetazolamide (0.1 mM) and 4,4'-diisothio cyanatostilbene-2,2'-disulphonic acid (DIDS, 0.5 mM) were used as specific inhibitors of carbonic anhydrase and NBC, respectively. The degree of inhibition was assessed by measuring the pHi recovery rate (${\Delta}pHi$/min) after cell acidification using an ammonium prepulse technique. In control experiments, ${\Delta}pHi$/min was $1.40{\pm}0.06$. Treatment of cells with 0.5 mM DIDS or 0.1 mM acetazolamide significantly reduced ${\Delta}pHi$/min to $1.14{\pm}0.14$ and $0.74{\pm}0.15$, respectively. Simultaneous application of DIDS and acetazolamide further reduced ${\Delta}pHi$/min to $0.47{\pm}0.10$. Therefore, DIDS and acetazolamide reduced ${\Delta}pHi$/min by 19% and 47%, respectively, while simultaneous application of both DIDS and acetazolamide caused a reduction in ${\Delta}pHi$/min of 67%. These results suggest that in addition to carbonic anhydrase, NBC also partially contributes to $HCO_3^-$ formation in human parotid gland acinar cells.

• Journal of Oral Medicine and Pain
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• v.32 no.1
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• pp.57-67
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• 2007

In currently, stress diseases are increased that present several sign and symptoms. Under stress condition, there are dry mouth, burning mouth syndrome, oral mucosa diseases and halitosis more frequently. Changing of salivary proportion is checked in almost patients with changing of function and structure in salivary gland. This study purpose are what effect stress does on salivary gland, and a-amylase on salivary gland. This study was resulted that 1. Under restraint stress, acinar cells are vacuolization and changing of intercellular spaces are separated, and peripheral tissues of duct are changed 2. Acinar cells were shrunk after 3 hours under restraint stress, intercellular space was separated after 6hours, peripheral tissues of duct started to change after 72 hours, and acinar cells and peripheral tissues of duct were all severely changed after 168hours. 3. In immunohistochemical study, amylase reaction was showed partially and irregularly after 3 hours, was getting little milder after 6 hours. And amylase reaction was gradually increased from the time of 12 hours after experiment up to the time of 48 hours after experiment. But after 168 hours, amylase appearance was diminished. According this result, emotional stress can change of salivary gland structure, and amylase secretion, the important digestive enzyme from salivary gland is changed and it is supposed to make digestive disorder and to make halitosis efficiency. So, we need to study about secretion of amylase.

• Applied Microscopy
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• v.37 no.2
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• pp.103-109
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• 2007

The ultrastructure of submandibular gland was examined in the Korean spider shrew, Sorex caecutiens. The submandibular gland wat composed of acini and salivary ducts. A submadibular acinus was a mixed gland having serous demilune cells and mucous cells that were filled with well developed rER, mitochondria and large amount of dense secretory granules. Serous acinar granules were oval shape without distinct limiting membrane on the border and it had only coarse specks with various density. Mucous acinar granules were oval shape without distinct limiting membrane and had a variety pattern with several thin or transparent bands into the homogeneous dense matrix. Thus submandibular acinar granules of S. caecutiens belonging to subfamily Soricinae were distinct from the other mammalian species including Crocidurinae, because of the absence of limiting membrane of acinar granules and specific pattern of mucous acinar granules. Granular duct cells had large amount of small granular vesicles and several characteristic structures of granule which were revered with stratified limiting membranes and filled with coarse serous-like granule or homogeneous matrix.