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PubMed2005

Expression and localization of chromogranin A gene and protein in human submandibular gland.

Human saliva chromogranin A (CgA) is clinically promising as a psychological stress marker. However, expression of CgA is poorly understood in humans, although salivary gland localization of CgA in other mammals, such as rodents and horses, has been demonstrated. In the present study, we investigated the expression and localization of CgA in the human submandibular gland (HSG) using various methods. CgA was consistently localized in serous and ductal cells in HSG, as detected by immunohistochemistry and in situhybridization. Reactivity was stronger in serous cells than in ductal cells. In addition, strong immunoreactivity for CgA was observed in the saliva matrix of ductal cavities. Western blotting gave one significant immunoreactive band of 68 kDa in the adrenal gland, HSG and saliva. Finally, CgA was detected in secretory granules of serous and ductal cells by immunoelectron microscopy. In conclusion, CgA in humans is produced by HSG and secreted into saliva.

Adrenal Glands↗
PubMed1995

Salivary secretion in cat submandibular gland mediated by chorda tympani afferents.

The aim of the present study was to investigate whether the afferent traffic from the tongue mediated only via the chorda tympani nerve (CTN) can still elicit reflex salivary and vasodilator responses in the cat submandibular gland (SMG) after section of the lingual nerve proper (LNP). Electrical stimulation of the chorda lingual nerve (CLN) at a site approximately 5 mm distal to the intersection of the CLN and the SMG duct elicited salivary and vasodilator responses in the SMG in sympathectomized cats. Both responses were unaffected by section of the LNP. The optimal frequency of CLN stimulation for submandibular salivation and vasodilation was 20 Hz, regardless of whether the LNP had been cut. Prior treatment with the autonomic ganglion blocker hexamethonium (10 mg/kg iv) virtually abolished the salivation and the blood flow increase in SMG. Prior treatment with scopolamine (0.1 mg/kg iv) almost abolished the salivary secretions but had no effect on the vasodilator responses in the SMG elicited by CLN stimulation after LNP section. The mechanism underlying the reflex submandibular salivation mediated by chorda tympani afferents appears to involve parasympathetic muscarinic receptors, but the mechanism for the vasodilator response has yet to be established. These results indicate that afferent traffic passing through the CTN on CLN stimulation is importantly involved in the parasympathetic reflex secretory and vasodilator responses in the cat SMG.

Adrenergic Antagonists↗
PubMed2005

Morphological alterations of submandibular glands caused by cisplatin in the rat.

Cisplatin is a platinum-containing antineoplastic agent, widely used in the treatment of various malignant neoplasms including head and neck cancer. In this study we examined ultrastructural changes of rat submandibular glands (SMGs) after cisplatin treatment. Male Wistar rats aged 8 weeks were used in the experiments. Cisplatin was injected intraperitoneally into the rats (5 mg/kg/day) for three consecutive days. After 1, 3, 5, and 7 days, the SMGs were removed and observed under an electron microscope. The terminal portion of the SMG is seromucosal. Acinar cells contained a parallel array of rough-surfaced endoplasmic reticulum in the basal cytoplasm, and the apical cytoplasm was filled with homogeneous secretory granules. Vacuolation of cells was observed on the first day after cisplatin treatment. On the third and fifth days, the rough-surfaced endoplasmic reticulum showed an irregular arrangement. Mitochondrial swelling, and the presence of autophagolysosomes was also confirmed. On the seventh day, the damage to organelles began to decrease and the SMG exhibited a tendency towards improvement.

Animals↗
PubMed1983

Stimulation of amylase and sialic acid releases from dog submandibular gland slices by pilocarpine or high K+ medium: a possible role of calmodulin for their releases.

The mechanism of amylase and sialic acid releases stimulated by pilocarpine or high K+ medium was investigated in the slices of dog submandibular glands. The release of both amylase and sialic acid was dose-dependently increased by pilocarpine and a considerable release was observed at pilocarpine concentrations of more than 1 microM. Similar effects were observed when K+ concentration in the medium was increased and the maximal response was observed at 75 mM K+. The release of amylase and sialic acid by pilocarpine or K+ considerably decreased by removing Ca2+ from the medium and the slices. The release of amylase in the Ca2+-deficient slices was nearly recovered by the addition of 2.5 and 5.0 mM Ca2+, whereas that of sialic acid was recovered by only 60-75%. Ca2+ inhibitors, La3+ and verapamil, and calmodulin inhibitors, trifluoperazine, prenylamine, and W-7, significantly inhibited the release of amylase and sialic acid induced by the stimulants. These results suggest that the release of amylase and sialic acid stimulated by pilocarpine or K+ is dependent on the presence of Ca2+, and that the activation of calmodulin is involved in the process of the release.

Amylases↗
PubMed1985

The streaming submandibular gland.

Twenty female young adult rats were injected with tritiated thymidine (3HTdR). The animals were then killed in groups of five, at the following times: 1 hour, and 4, 16, and 23 days. Autoradiograms of sections through the submandibular gland were prepared, and the location of labelled cells in relationship to tubuli and acini was recorded. The different tubular and acinar cross sections could be distinguished by their cell number. Narrow tubuli had fewer nuclei than the wider ones. The nuclear number of a cross section was defined as its class and the location of a labelled epithelial cell was expressed in relationship to the class where it was found. The location of a labelled stromal cell was determined by the class of its neighboring tubular or acinar cross sections. The mean cell numbers of intercalated, granular, and striated duct cross sections were, respectively, 4.7, 10.5, and 10.2, while the average cell content of acini was 4.7 cells. One hour after labelling most labelled tubular epithelial and stromal cells were found in tubular cross sections (or low tubular classes), while in the acini, labelled epithelial and stromal cells were found mainly in wider cross sections (or higher acinar classes). Within the next 23 days labelled tubular cells and stroma proceeded into higher classes, while labelled acinar epithelium and stroma cells were displaced into narrower cross sections (or lower classes). The displaced tubular epithelium and stroma covered daily 0.26 classes. At this velocity the cell will reach the highest tubular class in 62 days and the estimated maximal tubular cell life span is 62 days.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2006

Expression and characteristics of vanilloid receptor 1 in the rabbit submandibular gland.

Vanilloid receptor 1 (VR1) is a polymodal receptor originally found in sensory neurons of the central nervous system. Recent evidence indicates that VR1 is also expressed in non-neuronal tissues. We report here endogenous expression of VR1 in rabbit submandibular gland (SMG) and its possible role in regulating saliva secretion based on: (i) the expression of VR1 mRNA and protein detected in SMG; (ii) VR1 was mainly localized in the basolateral membrane of duct cells and the cytoplasm of acinar cells and also in cytoplasm of primary cultured neonatal rabbit SMG cells; (iii) stimulation of neonatal rabbit SMG cells with capsaicin induced a significant increase in intracellular calcium, and capsazepine, a VR1 antagonist, abolished this increase; (iv) infusion of capsaicin via the external carotid artery to isolated SMG increased saliva secretion of the gland. These findings indicated that VR1 was expressed in SMG and appeared to play an important role in regulating saliva secretion.

Animals↗
PubMed1990

Histochemical studies on irradiated and obstructive human submandibular glands. Protein distribution and lectin-binding in the degenerative glands.

Immunohistochemical protein distribution of alpha-amylase (Am), lysozyme (Ly), cytokeratinin (CK), S-100 protein (S-100) and secretory component (SC), and lectin-binding (SBA and UEA-I) profiles were studied in 10 obstructive and 20 irradiated human submandibular glands which were surgically extirpated. Degenerative intensity of the glands was graded as I, II and III based on the order of severity. All proteins generally existed in serous acinic cells of the intact glands. The proteins immunoreactivities became weak even in mildly inflamed glands (grade I), and nearly disappeared from the moderately damaged glands (grade II). Duct cells had clear CK and some cells reacted with the anti-SC antibody, but other proteins were not observed on the ducts. Mucous cells possessed none of the proteins, and their lectin-binding was only traceable in some glands. Compared with immunoreactivities in the proteins, lectin-binding profiles were different. SBA and UEA-I bound somewhat similarly to both acinic and duct cells, and the binding was hardly affected even by severe degeneration (grade III). Between obstructive and irradiated glands, no obvious difference was observed in either protein distribution or lectin-binding. From the above, it seems that some proteins are more affective to the degeneration and that lectin-binding sugar residues are non-affective against the degenerative changes of the tissues.

Histocytochemistry↗
PubMed1990

Substrate specificity of prorenin converting enzyme of mouse submandibular gland. Analysis using site-directed mutagenesis.

Renin is produced from a larger, inactive precursor, prorenin, through endoproteolytic cleavage at paired basic amino acids. Recently, we have purified and characterized an enzyme, which catalyzes the endoproteolytic process, from mouse submandibular gland. The enzyme, named prorenin converting enzyme, specifically cleaves the peptide bond on the COOH-side of the Arg residue at the Lys-Arg pair of mouse Ren 2 prorenin, but does not cleave mouse Ren 1 and human prorenins. In this study, by synthesizing a series of mutant mouse prorenins using site-directed mutagenesis and the Xenopus oocyte expression system, we have investigated the role of the basic pair as the recognition signal for the enzyme as well as the determinant of the substrate specificity. The results indicate that the basic amino acid at the COOH-side but not at the NH2-side of the basic pair of Ren 2 prorenin is essential for processing directed by prorenin converting enzyme, and that the Arg residue at the COOH-side is more preferable for processing than the Lys. The results also demonstrated that the presence of a Pro residue next to the Lys-Arg pair prevents the processing of Ren 1 prorenin.

Amino Acid Sequence↗
PubMed1993

Effects of female hormones and 3,5,3'-triiodothyronine or dexamethasone on induction of epidermal growth factor and proteinases F, D, A, and P in the submandibular glands of hypophysectomized male mice.

The effects of progesterone (Pro), 5 alpha-dihydrotestosterone (DHT), 17 beta-estradiol (E2), T3, and dexamethasone (Dex), given alone or in combination, on induction of epidermal growth factor (EGF) and proteinase isozymes in the submandibular glands of hypophysectomized mice were examined. Each hormone, except E2, acting alone had essentially comparable inductive effects on EGF concentrations and on total proteinase activity. E2 alone had no inductive effect at all. Pro acted synergistically with T3, but its inductive effect was diminished when given with Dex. E2 was not synergistic with T2, and it inhibited the effect of Dex; it also partially blocked the action of DHT on induction of proteinase activity but not of EGF. Simultaneous administration of all five hormones restored total proteinase activity completely but EGF levels to only 50% of values for intact males, respectively. Submandibular proteinases were resolved by isoelectric focusing into four isozymes: proteinase F (pI 4.8), proteinase D (pI 5.8), proteinase A (pI 6.2), and proteinase P (pI 10.0). Pro alone slightly increased levels of proteinase F but greatly raised levels of the other three isozymes. This inductive action was augmented when Pro was given with T3 but blocked when it was given with Dex. E2 alone not only failed to induce any of the isozymes, but even further reduced levels of proteinase F. It also decreased the inductive effects of T3 on these isozymes, and with Dex completely blocked induction of proteinases D, A, and P. E2 plus DHT suppressed proteinase F levels and only induced proteinase A. All five hormones together reestablished the isozyme profile seen in intact males. These results show that Pro by itself is as capable as androgens, thyroid hormone, or glucocorticoid in regulating expression of these submandibular polypeptides, and that its action can be modulated by other pituitary-dependent hormones. In addition, they demonstrate that E2 does not regulate their expression, and that it has an inhibitory effect on the inductive action of other hormones. Last, they indicate that these various hormones may regulate expression of EGF and of each of the individual proteinase isozymes differently.

Animals↗
PubMed1989

Postnatal changes in an alpha subunit isoform, alpha(S), of Na+,K(+)-ATPase in the submandibular gland of rats.

The alpha and alpha(+) isoforms of Na+,K(+)-ATPase were isolated from the kidney and brain of rats and purified. Their antisera were raised to analyze the alpha isoforms in rat tissues. We found that the submandibular gland (SMG) contains a new immunoreactive alpha subunit isoform, designated alpha(S) in this report, in addition to alpha identical with those found in the kidney or brain. The new alpha(S) strongly reacted with anti-alpha-antiserum but to a much lesser extent with anti-alpha(+)-antiserum. The alpha(S) had a slightly lower molecular weight (approximately 90,000) than the brain and kidney alpha isoforms. Various fractions of SMG tissues were added to the SMG microsomes and incubated in order to test whether or not the alpha(S) is formed artificially; no increase of alpha(S) was observed by these treatments, suggesting that the alpha(S) was not the product formed from alpha during the preparation of microsome sample, but was rather a protein originally present in the SMG. The alpha(S) protein was not detected in the SMG of 2- or 5-week-old rats, but it gradually increased in rats older than 8 weeks, reaching the maximum in 30-week-old animals. The Na+,K(+)-ATPase activity in the SMG increased concomitantly with the increase of alpha(S), indicating that Na+,K(+)-ATPase comprising alpha(S) also shows enzyme activity; it is speculated that alpha(S) may have some unique and unknown function(s) in older rats.

Animals↗
PubMed2006

Peroxidase secretion in rat submandibular glands induced by PGE2: role of cAMP and nitric oxide.

Free radicals are associated with the appearance of disorders such as tumours, CNS alterations and inflammatory pathologies. Their levels are known to be increased in inflammatory diseases due to the activity of prostaglandins, which are related to protein secretion including enzymes. Peroxidase is an oral enzyme that is implicated in the defence of the oral cavity. In this paper, investigations of the effect and mechanism of the activity of prostaglandin E2 (PGE2) on peroxidase secretion of female rat submandibular glands are reported. Results showed that PGE2 significantly increased the secretion of submandibular peroxidase and that this effect was mediated by an increase of intracellular cAMP and nitric oxide synthase activation. This could imply that prostaglandins play a modulatory role in diseases where free radicals are involved.

Animals↗
PubMed1996

Microsomal diacylglycerol acyltransferase in rat parotid and submandibular glands: acylation of 1,2-dioleoyl-sn-glycerol dispersed with phospholipids.

In order to investigate microsomal diacylglycerol acyltransferase activity, ethanol or several detergents have been used as a dispersing agent for water-insoluble substrates. However, ethanol acyltransferase interferes with the activity of this enzyme, and detergents inhibit it. We examined the properties of microsomal diacylglycerol acyltransferase in rat salivary glands without detergents or organic solvents. 1,2-Dioleoyl-sn-glycerol (1,2-diolein) was dispersed by sonication. The activity was measured as the formation rate of [14C]triglyceride using [1-14C]palmitoyl-CoA as an acyl-donor. The reaction was dependent on the microsomal protein and 1,2-diolein at least up to 145 micrograms/ml and 3.6 mM, respectively. The specific activities were 3.91 +/- 0.57 and 3.80 +/- 0.77 nmol/min per mg protein (SEM, n = 4) in the parotid and submandibular glands, respectively. They were 12- to 20-fold higher than the activities in liver, brain and spleen, and two orders of magnitude higher than that assayed with microsomal endogenous diacylglycerol. Adding tissue phospholipids to 1,2-diolein suspension reduced the concentration of 1,2-diolein required for the maximal velocity. A similar, but reduced, effect was induced by egg yolk phosphatidylcholine in place of the tissue phospholipids. The level of activity was recovered by adding another phospholipid class to the phosphatidylcholine. The results suggested that the physical condition of the substrate diacylglycerol affects diacylglycerol acyltransferase activity in rat salivary gland microsomes.

Acyltransferases↗
PubMed2003

Dose-dependent morphological changes of intercellular canaliculi during stimulation with carbachol and isoproterenol in the isolated rat submandibular gland.

Intercellular canaliculi (IC) form a primary mixing reservoir for transcellularly and paracellularly secreted saliva whose composition depends on the degree of elevation of cytosolic Ca2+ and of cytosolic cyclic AMP concentrations caused by the secretagogues employed. In perfused rat submandibular gland (SMG), appearance of exocytosis on IC reflected the quantity of secreted mucin. Morphological observations were carried out by HR-SEM using a modified osmium maceration method on specimens treated with CCh and/or ISP. Mild secretory stimulation revealed that exocytosis did not occur simultaneously, even along the same intercellular canaliculus. Higher doses did not alter the spatial distribution of exocytosis along intercellular canaliculi but increased its temporal frequency, dose dependently. These findings lead us to conclude that, under low levels of secretory stimulation, exocytosis does not show a dose-dependent change, but that its spatial and temporal frequency changes in a dose-dependent manner.

Adrenergic beta-Agonists↗
PubMed1962

Action of some ganglion-stimulating substances on the secretion of saliva from the submandibular gland.

An attempt has been made to determine whether the ganglionic actions of pilocarpine and of 4-(m-chlorophenylcarbamoyloxy)-2-butynyltrimethylammonium chloride (McN-A-343) contribute to their effect on salivary secretion. Salivary flow was measured from the submandibular glands of spinal cats. Destruction of the superior cervical ganglion and adrenalectomy failed to reduce the stimulant effect of pilocarpine and McN-A-343. Substances known to interfere with the ganglionic actions of pilocarpine (cocaine, methadone and choline 2:6-xylyl ether bromide) likewise failed to modify the response. It is concluded that stimulation of autonomic ganglia and of the adrenal medulla does not contribute to the salivary secretion observed after intravenous injections of pilocarpine and of McN-A-343. Dimethylphenylpiperazinium, a nicotine-like ganglion-stimulating substance, causes salivary flow by stimulating the adrenal medulla as well as parasympathetic ganglion cells; stimulation of the superior cervical ganglion by this substance does not contribute to the salivary response.

(4-(m-Chlorophenylcarbamoyloxy)-2-butynyl)trimethy↗
PubMed2000

Comparative image analysis of EGF immunoreaction in rat submandibular gland using 3,3'-diaminobenzidine with metal enhancer substrate.

We have shown the efficacy of image analysis using 3,3'-diaminobenzidine (DAB) with a metal enhancer substrate for demonstrating quantitative differences in the amount of epidermal growth factor (EGF) in the submandibular gland from normal, castrated, and testosterone propionate (TP) treated castrated rats. Immunohistochemical determination of EGF visualized by DAB-nickel reagent was performed with image analysis using a computed image analyzer system (ACAS 570). Immunohistochemistry for EGF disclosed positive staining in granular convoluted tubule cells in the tissue sections from each experimental group. Using the tools of a competent data program installed in the ACAS 570 software, we measured quantitative differences among the experimental glands examined. Castration was shown to elicit a significant reduction in the EGF-positive area and staining intensity, and administration of TP to the castrated animals restored these parameters to levels greater than those of normal rats. Our study demonstrates that a simple, inexpensive, commercially available metal enhancer substrate can be applied accurately to the computer assisted quantification of histochemical hormone-induction studies.

3,3'-Diaminobenzidine↗
PubMedDate not supplied

Effects of castration and thyroidectomy on expression of lectin-binding sugar moieties and endolectins in mouse submandibular glands. A glycohistochemical study.

The aims of this study were to assess histochemically by means of biotinylated lectins and neoglycoproteins the effects of castration and thyroidectomy on the expression of carbohydrate moieties and endogenous sugar-binding proteins (endolectins), respectively, in submandibular glands of mice. Characteristic binding patterns depending upon the type of lectin were observed primarily in the granular convoluted tubules. For the first time carbohydrate-binding sites were also localized in this glandular tissue. Moreover, the endocrine status was found to affect the histochemical expression of sugar moieties of glycoconjugates as well as endolectins, thus modifying both sides of presumed protein-carbohydrate interactions. Further work is needed to identify these molecules and to understand better their endogenous roles.

Animals↗