Heat-induced changes in the rat parotid gland

Gland size, amylase activity, total proteins and DNA concentrations were measured in parotid gland during chronic heat exposure (34 degrees C). Subcellular fractionation was performed. Exposure to heat resulted in a decrease in gland size and in DNA concentration; amylase activity/100 mg tissue and amylase specific activity to protein concentration ratio increased. Subcellular fractions, except plasma membrane, decreased significantly. Thus, chronic heat exposure slows down the production of gland constituent proteins, whereas acinar function, e.g. the production and storage of exportable proteins, is not disturbed. The reduced gland size is apparently due to hypoplasia.     

Electrophoretic identification of calmodulin in rat parotid gland

A heat-stable extract of rat parotid gland contained a protein the electrophoretic migration of which on SDS-polyacrylamide gel was accelerated by Ca2+. Its apparent molecular weight was about 17 or 20 K in the presence or absence of Ca2+, respectively; it was able to bind Ca2+. These properties are exactly those of purified calmodulin.     

Terbutaline-induced enlargement of the rat parotid gland

The β-sympathomimetic drug terbutaline was shown to be a powerful secretogogue for the rat parotid gland. It was also found to be capable of causing a transient enlargement of the parotid which was similar to that produced by isoproterenol.     

Methotrexate-induced changes in rat parotid and submandibular gland function

Methotrexate (MTX) was injected intraperitoneally at a dose of 15 mg/kg into adult rats daily for three consecutive days. When compared with saline-injected, ad libitum-fed rats or pair-fed saline-injected rats, MTX decreased unstimulated parotid gland weight, RNA content, and amylase content. RNA content was also decreased in submandibular glands. The parotid and submandibular gland outputs of sodium and chloride, but not that of potassium, were decreased following MTX injection. Ductal perfusion demonstrated decreased net efflux of sodium and chloride from the excretory duct of the submandibular gland. The effects of MTX on unstimulated parotid gland weight, RNA content, and amylase content are consistent with its ability to inhibit protein synthesis through depletion of folate co-factors. However, the mechanism by which the alterations in ion transport occurred is not clear at this time.     

Effect of ionizing radiation on rat parotid gland

A common side effect of radiotherapy used in the treatment of oral cancer is the occurrence of structural and physiological alterations of the salivary glands due to exposure to ionizing radiation, as demonstrated by conditions such as decreased salivary flow. The present study evaluated ultrastructural alterations in the parotid glands of rats receiving a fractionated dose (1,500-cGy) of radiation emitted by a Cesium-137 source and rats that were not subjected to ionizing radiation. After sacrifice, the parotid glands were removed and examined by transmission electron microscopy. Damage such as cytoplasmic vacuolization, dilatation of the endoplasmic reticulum and destruction of mitochondria, as well as damage to the cellular membrane of acinar cells, were observed. These findings lead to the conclusion that ionizing radiation promotes alterations in the glandular parenchyma, and that these alterations are directly related to the dose level of absorbed radiation. Certain phenomena that appear in the cytoplasm and nuclear material indicate that ionizing radiation causes acinar cell death (apoptosis).     

Effects of zinc deficiency on rat parotid gland

Hitherto, the effect of zinc deficiency on the rat parotid gland was not known. Male rats were fed either a low zinc diet (0.4-0.8 ppm Zn) or a zinc adequate diet (approximately 40 ppm Zn) for 35-42 days. The experimental animals exhibited retarded body growth, anorexia, loss of hair and decreases in plasma and hair zinc levels. In comparison to the ad libitum controls, the parotid glands of the pair-fed group showed a decrease in cell number, but an increase in cell mass, protein and amylase content. In the parotid glands of the experimental group, cell number was comparable to the pair-fed group, but there was a decrease in cell mass and protein content as compared to the pair-fed group. These findings suggest that zinc deficiency and its associated reduced food intake impair growth of the parotid gland and, further, that zinc deficiency per se appears to lead to a diminution in gland protein synthesis. It is proposed that a compromise in parotid gland function could be a contributory factor underlying the increased susceptibility to dental cares known to occur in zinc deficiency.     

Effects of zinc deficiency on rat parotid gland

Hitherto, the effect of zinc deficiency on the rat parotid gland was not known. Male rats were fed cither a low zinc diet (0.4–0.8 ppm Zn) or a zinc adequate diet (approximately 40 ppm Zn) for 35–42 days. The experimental animals exhibited retarded body growth, anorexia, loss of hair and decreases in plasma and hair zinc levels. In comparison to the ad libitum controls, the parotid glands of the pair-fed group showed a decrease in cell number, but an increase in cell mass, protein and amykisc content. In the parotid glands of the experimental group, cell number was comparable to the pair-fed group, but there was a decrease in cell mass and protein content as compared to the pair-fed group. These findings suggest that zinc deficiency and its associated reduced food intake impair growth of the parotid gland and, further, that zinc deficiency per se appears to lead to a diminution in gland protein synthesis. It is proposed that a compromise in parotid gland function could be a contributory factor underlying the increased susceptibility to dental caries known to occur in zinc deficiency.     

Transglutaminase expression in rat parotid gland after isoproterenol stimulation

Transglutaminases (E.C. 2.3.2.13) are calcium-dependent enzymes that catalyze the covalent cross-linking of proteins, and occur in multiple molecular forms in a variety of tissues. Distribution of each form of transglutaminase varies with different tissues. Studies were undertaken to characterize the form of transglutaminase expressed in rat parotid gland, and to examine a possible physiological role for the enzyme. It was found that chronic treatment of rats with the beta-adrenergic agonist isoproterenol (IPR) resulted in the induction of parotid transglutaminase activity. The properties of this transglutaminase appeared to be distinct from those of the well-characterized guinea pig liver cytosol transglutaminase (TGase C). The findings that protein polymerization (observed on SDS-PAGE) and incorporation of radioactive putrescine, a polyamine, into protein occur in the presence of exogenous transglutaminase and calcium indicated that certain rat parotid salivary proteins are or could be substrates for this enzyme. Analysis of proteolytic digests of rat parotid salivary proteins on an amino acid analyzer and by high-performance liquid chromatography also indicated that these salivary proteins contain gamma-glutamyl derivatives of primary amines (e.g., polyamines or lysine), post-translational products of transglutaminase catalysis. The possible physiological function of this enzyme in the oral cavity might be stabilization of proteinaceous structures during normal oral homeostasis and/or woundhealing.     

Calcium uptake in rat parotid gland secretory granules.

45Ca2+ uptake in isolated rat parotid secretory granules was examined in the presence of oxalate. Uptake of calcium was dependent on time, with the maximum occurring at 15 min. The uptake of calcium was dependent on adenosine-5'-triphosphate (ATP), and substitution of ATP with beta, gamma-methylene-ATP did not stimulate calcium uptake. Enzyme marker analysis indicated that mitochondria accounted for no greater than 3.0 +/- 0.2% of the observed ATP-dependent calcium uptake. Calcium uptake was blocked by the ATPase inhibitors tributyltin, IC50 = 12.2 +/- 0.6 nmol/L and 4-acetamido-4'-isothiocyano-2,2'-stilbene disulphonic acid (SITS), IC50 = 3.0 +/- 0.3 mumol/L. These results indicate that in the parotid secretory granule there is a calcium uptake mechanism that is dependent on the hydrolysis of ATP and is suppressed by two inhibitors of granule ATPase.     

Characterization of nitric oxide synthase in the rat parotid gland

Nitric oxide (NO) acts as an inter- and intracellular signalling molecule of various cells such as vascular endothelium, macrophages, and neurones. NO is produced by nitric oxide synthase (NOS) from L-arginine. Here the characteristics of NOS in the rat parotid gland were investigated. Approximately 74% of total activity of NOS was present in the cytosolic fraction. For full activation of the NOS in the cytosolic fraction, tetrahydroxybiopterin, NADPH, Ca(2+) and calmodulin were needed as cofactors, because the activity was clearly reduced in the absence of tetrahydroxybiopterin, NADPH, or Ca(2+), or in the absence of calmodulin and presence of trifluoperazine, a calmodulin antagonist, in the reaction mixture. The partially purified NOS activity was completely abolished in the absence of calmodulin or Ca(2+), and activated by them in a dose-dependent manner; EC(50) for calmodulin and Ca(2+) were 10 and 340 nM, respectively. The K(m) for L-arginine was 1.57 microM. Immunoblot analysis revealed that a 165-kDa protein band in the rat parotid gland cytosolic fraction cross-reacted with a rabbit polyclonal antibody against human brain NOS. These results suggest that NOS of the rat parotid gland is a neuronal isoform and that its activity is regulated by physiological concentrations of calmodulin and Ca(2+).     

Sedimentation profiles of rat parotid gland polysomes in sucrose gradients

Fractionation of rat parotid gland in a sucrose -TKM medium, pH 7.6, leads to extensive disaggregation of polyribosomes by the high level of endogenous, free RNase present in the tissue. Previous studies have shown that prior administration of a potent sialogogue to the animals before sacrifice profoundly reduces the tissue RNase activity, thus permitting isolation of ribonucleoproteins which exhibit active protein synthesis in a cell-free system. This report indicates a similar reduction of active RNase in parotid tissue by the crude enzyme inhibitor present in rat liver postmicrosomal supernatant, and also shows that the size distribution of parotid ribosomal aggregates in sucrose gradients in the presence of this inhibitor parallels findings in animals treated with isoproterenol. This technique will be useful in studying shifts in parotid polysome profiles under conditions which affect protein synthesis in the organ and eliminates the need for the use of potent sialogogues whose effects on the parameters under investigation are not fully known.     

Inhibition of Streptococcus mutans biofilm formation using extracts from Assam tea compared to green tea

ObjectiveStreptococcus mutans, a gram-positive oral bacterium, has been identified as one of the principal etiological agents of human dental caries. To clarify the nature of the difference anti-biofilm effect against S. mutans between Assam tea from Camellia sinensis var. assamica, partially fermented, and green tea from Camellia sinensis, non-fermented, active agents from the teas were purified.MethodsEffects of Assam tea and green tea samples on biofilm were assessed by using the conventional titer plate method and the human saliva-coated hydroxyapatite discs. The purification and identification of inhibitors were performed by using ultrafiltration with centrifugal filter devices and high performance liquid chromatography.ResultsAssam tea has stronger biofilm inhibition activity against S. mutans than green tea. A substance of <10 kDa in mass in Assam tea had a high concentration of galloylated catechins and a stronger biofilm inhibiting activity than green tea. In contrast, substances >10 kDa in mass from green tea included higher concentrations of polysaccharides composed of galacturonic acid, such as pectin, that enhance biofilm formation.ConclusionsThe higher concentrations of galloylated catechins in Assam tea may assist in prevention of dental caries, whereas in green tea, this mode of inhibition was likely offset by the presence of pectin. Purification of catechins in partially fermented Assam tea with lower-molecular-weight polysaccharide than pectin may be useful for developing oral care products such as toothpaste and oral care gel pastes.     

Chlorpromazine inhibition of muscarinic-cholinergic responses in the rat parotid gland

The ability of chlorpromazine (CPZ) to inhibit muscarinic-cholinergic secretory events was studied in vitro in rat parotid acinar cells. CPZ inhibited carbachol-induced amylase release in a dose-dependent fashion but had no effect on that elicited by isoproterenol. The inhibition of parotid protein synthesis induced by carbachol, but not that induced by A23187, was blocked by CPZ. CPZ exhibited a dose-dependent inhibition of [3H] quinuclidinyl benzilate (QNB) binding to muscarinic receptors, and altered the KD of the receptor for the ligand. These results are consistent with an ability of CPZ to inhibit muscarinic-cholinergic-induced salivary secretion by complex interference with receptor binding. In addition, CPZ may block parotid-muscarinic responses by impeding a post-receptor signaling step which is proximal to Ca2+ mobilization.     

Isoproterenol-induced changes in glutathione metabolism in the rat parotid gland

Rat parotid glands were removed at timed intervals after intraperitoneal injection of isoproterenol (IPR). Reduced/oxidized glutathione ratios, glutathione peroxidase, glutathione reductase and amylase activities were determined. Parotid gland amylase content fell to less than 5 per cent of control values within 1 h after IPR, then slowly returned to control levels by 24 h. The ratio of reduced/oxidized glutathione was decreased at 2 h after IPR injection, returned to near control values at 6 h, then increased above control levels at 9, 12 and 24 h. Glutathione peroxidase activity was increased only at 12 h after injection of IPR, whereas glutathione reductase activity initially decreased, then reached peak values at 12 h. These results indicate that alterations in metabolic activity during the secretory cycle are accompanied by changes in the reduced/oxidized glutathione ratio and the activity of glutathione-metabolizing enzymes. Cellular events contributing to these alterations in glutathione metabolism may include exocytosis, endocytosis and membrane recycling, increased amino-acid transport and resynthesis of secretory proteins.