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PubMed1999

Regulation of the Na+-K+(NH4+)-2Cl- cotransporter of rat submandibular glands.

A cellular suspension from rat submandibular glands was exposed to different concentrations of NH4Cl, and the variations of the intracellular concentration of calcium ([Ca2+]i) and the intracellular pH (pHi) were measured using fura-2 and 2',7'-bis-(2-carboxy-ethyl)-5(6)-carboxyfluorescein. More than 5 mmol/l NH4Cl significantly increased the [Ca2+]i without affecting the response to 100 micromol/l carbachol. When exposed to 1 and 5 mmol/l NH4Cl, the cells acidified immediately. At 30 mmol/l, NH4Cl first alkalinized the cells and the pHi subsequently dropped. This drop reflects the uptake of NH4+ ions that dissociate to NH3 and H+ in the cytosol. These protons are exchanged for extracellular sodium by the Na+/H+ exchanger because the presence of an inhibitor of the exchanger in the medium increased the acidification induced by 1 mmol/l NH4Cl. Ouabain partly blocked the uptake of NH4+. In the combined presence of ouabain and bumetanide (an inhibitor of the Na+-K+-2Cl- cotransporter), 1 mmol/l NH4Cl alkalinized the cells. The contribution of the Na/K ATPase and the Na+-K+-2Cl- cotransporter in the uptake of NH4+ was independent of the presence of calcium in the medium. Isoproterenol increased the uptake of NH4+ by the cotransporter. Conversely, 1 mmol/l extracellular ATP blocked the basal uptake of NH4+ by the cotransporter. This inhibition was reversed by extracellular magnesium or Coomassie Blue. It was mimicked by benzoyl-ATP but not by CTP, GTP, UTP, ADP, or ADPbetaS. ATP only slightly inhibited the increase of cyclic AMP (-22%) by isoproterenol but fully blocked the stimulation of the cotransporter by the beta-adrenergic agonist. ATP increased the release of 3H-arachidonic acid from prelabeled cells but SK&F 96365, an imidazole-based cytochrome P450 inhibitor, did not affect the inhibition by ATP. It is concluded that the activation of a purinoceptor inhibits the basal and the cyclic AMP-stimulated activity of the Na+-K+-2Cl- cotransporter.

Adenosine Triphosphate↗

PubMed1980

Pharmacological analysis of salivary and blood flow responses to histamine of the submandibular gland of the dog.

1 The submandibular gland in situ was perfused with blood through the glandular artery at constant pressure in anaesthetized dogs. Drugs were administered intra-arterially. 2 Histamine produced both salivation and an increase in blood flow, each response having an early and a late component. 3 Marked tachyphylaxis to histamine developed in both of the salivary responses but only in the late blood flow response to histamine. 4 The early and late salivary responses were abolished and the late blood flow response was diminished by infusion of tetrodotoxin in doses that abolished the salivary and blood flow responses to electrical stimulation of the chorda-lingual nerve. 5 The whole salivary response to histamine was abolished by infusion of (--)-hyoscyamine in doses that greatly antagonized the salivary and blood flow responses to acetylcholine, whereas the blood flow responses to histamine were scarcely modified. These doses of (--)-hyoscyamine abolished the salivary response to chorda-lingual nerve stimulation but left the blood flow response to it unaffected. 6 The salivary and blood flow responses to histamine were unaffected by infusion of hexamethonium in doses that almost abolished the salivary and blood flow responses to chorda-lingual nerve stimulation. 7 The whole salivary response to histamine was abolished and the late blood response to histamine was partially inhibited by the histamine H1-receptor antagonist, mepyramine, but not by the histamine H2-receptor antagonist, metiamide. 8 The early blood flow response to histamine was antagonized by both mepyramine and metiamide but mepyramine was far more effective than metiamide. 9 These results led to the following conclusions: (1) the whole salivary response and a part of the late blood flow response to histamine are due entirely to excitation of parasympathetic postganglionic neurones; (2) neuronal histamine receptors involved are exclusively of the H1-type; (3) histamine has no direct stimulant action on the glandular cells; (4) the early blood flow response and the remaining part of the late blood flow response to histamine result from the direct action on vascular smooth muscle in the glandular vascular bed; (5) vascular histamine receptors consist of H1- and H2-receptors.

Animals↗

PubMed1994

Epithelial class II MHC antigen normally expressed by submandibular glands of elderly people.

Immunohistochemical studies were made on 81 submandibular glands obtained from over 60-year-old autopsy cases. Those patients who had had either sialolithiasis or autoimmune diseases were not included in this study. HLA-DR expression was observed in the great majority (95.1%) of the aged submandibular glands, especially on the serous acinar cells and the intercalated duct cells, regardless of the appearance of focal lymphocytic infiltration. The incidence of HLA-DP expression was slightly lower (85.2%) and HLA-DQ was much lower (42.0%) than that of HLA-DR. These data support the proposal that epithelial expression of class II antigens in aged glands is a focal sign of age-related immunological disorders.

Aged↗

PubMed2006

Long-term survival of an allografted submandibular gland in a miniature swine model given immunosuppressant drugs.

We used a model of allografts of submandibular glands in miniature swine to test the requirements of immunosuppressants for the survival of such grafts. Animals in the first group (n=6) were given no immunosuppressant, and the submandibular glands were rejected within 7 days. Those in the low-dose group (n=4) were treated with a low dose of cyclosporine, methylprednisolone, and azathioprine. The allografts were rejected on days 15, 17, 19, and 20. The animals in the high-dose group (n=6) were treated with a high dose of cyclosporin, methylprednisolone, and azathioprine. Three allografts were rejected, on days 35, 48, and 60. One animal died from anaesthetic problems on day 30 after transplantation, and there were no signs of rejection in the allograft. The other two allografts survived for 100 days with secretory function and no signs of rejection. After we discontinued the immunosuppression on day 100, the two remaining allografts were rejected on days 121 and 128. Blood tests and biopsy specimens of the heart, lung, liver, and kidneys were normal in all animals that were given immunosuppressants. To our knowledge, this study is the first to describe long-term survival of allografted submandibular glands with secretory function in miniature swine.

Animals↗

PubMed1995

Histochemical localization of succinate dehydrogenase in intracerebral mouse submandibular gland transplants.

Histochemical localization of succinate dehydrogenase in developing intracerebral auto- and homotransplants of the mouse submandibular gland was investigated in the course of a 5-month period after transplantation. Eight weeks after grafting, the enzyme pattern in the parenchyma of homotransplants was comparable with the histochemical picture of a fully mature submandibular gland in situ. At this time, numerous acini showed a weak activity, very numerous striated ducts a strong activity, and less frequent developing convoluted granular tubules slightly weaker activity, than in the striated ducts. Beginning histochemical differentiation of convoluted granular tubules was noted only in homotransplants, located in the brain of male recipients. On the other hand, only a weak activity in the cytoplasm of non-differentiated duct--like structures of some autotransplants was seen. Homotransplants of non-differentiated submandibular gland of newborn donors were found to be a more suitable transplantation object capable of postnatal development of gland parenchyma than autografts of fully differentiated gland of adult animals that did not enter the cytodifferentiation stage.

Animals↗

PubMed1991

Histochemistry of sympathetic neurons allotransplanted from young and aged mice to the submandibular gland.

Sympathetic ganglion tissue of young (3 months) and aged (24 months) NMRI mice was allotransplanted into the submandibular gland to study the influence of aging on the survival of grafted neurons. The submandibular gland (SMG) was chosen as a host tissue because of its high concentration of NGF and good blood supply. Four weeks postgrafting the viability of transplants was evaluated using the formaldehyde-induced fluorescence technique, tyrosine hydroxylase (TH) immunohistochemistry, and morphometry. The density of neurons, catecholamine fluorescence and TH immunoreactivity (TH-IR) appeared to be almost unchanged when the transplant was completely surrounded by the SMG tissue, whereas transplants located within the interlobular septum and capsule, or even outside the capsule, showed significantly reduced neuronal survival. The shape of most of the transplanted neurons was not different from those in the intact ganglia. The average diameter of the transplanted young neurons was significantly decreased; this was not the case with the aged neurons. The histograms of grouped diameter values showed a shift to smaller cells in ganglion transplants in both age groups. The transplants in mice treated with 6-OH-dopamine showed considerable regrowth of adrenergic nerve fibers. There seemed to be no marked difference in the survival of transplanted neurons between aged and young animals. The results indicate that the sympathetic neurons from both young adult and aged animals survive the allotransplantation procedure. The neurotrophic factors together with dense vascularization present in the mouse submandibular gland may be beneficial for the restoration of the integrity of mature and aged adrenergic neurons.

Aging↗

PubMed2005

Epidermal cyst of submandibular gland.

The differential diagnosis of a cystic lesion in the submandibular area can be difficult. We report a case of epidermal cyst of submandibular gland which is relatively rare compared to the commoner epidermoid cyst.

Adult↗

PubMed2000

Immunohistochemical localization of carbonic anhydrases I, II, and VI in the developing rat sublingual and submandibular glands.

Carbonic anhydrase has been localized to the acini and ducts of mature rat salivary glands. This enzyme has been associated with ion transport, a prominent function of striated and excretory ducts in salivary glands, suggesting that it might be used as a marker of ductal differentiation. The purpose of this study was to immunohistochemically document developmental changes in carbonic anhydrase in the ducts of the rat sublingual and submandibular glands. Immunohistochemistry was performed with antibodies to human carbonic anhydrase isoenzymes I, II and VI on sections of sublingual and submandibular glands from rats at representative postnatal developmental ages. Reactions were weak in the ducts of both glands at 1 day, then progressively increased. By 42 days, reactions had the adult pattern of virtually none in the mucous or seromucous acini, moderate to strong in the striated and excretory ducts, and none to weak in the intercalated ducts. Weak to moderate reactions were observed in the granular convoluted tubules of the submandibular gland as they became recognizable at age 42 days. Reactions to carbonic anhydrase I and II antibodies also increased from none (1 day) to modest (42 days) in the demilunes of the sublingual gland. The order of reaction intensity of the antibodies was II > I > VI. When localized via these anti-human antibodies, carbonic anhydrase is a useful marker of the functional differentiation of the striated and excretory ducts of the developing rat sublingual and submandibular glands.

Animals↗

PubMed1989

Defective phosphorylation of a calmodulin-binding protein in cystic-fibrosis submandibular glands.

Calmodulin-binding proteins in fractions purified from human submandibular glands by calmodulin-Sepharose were phosphorylated with [gamma-32P]ATP, in the absence of exogenous protein kinase. The major proteins phosphorylated had molecular masses of 45, 51 and 61 kDa. Phosphorylation was increased by activators of protein kinase C and inhibited by H-7. Phosphorylation of the 61 kDa band was markedly decreased in cystic-fibrosis submandibular glands.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

PubMed1985

Effects of the selective beta 2-adrenergic agonists, procaterole and terbutaline, on protein secretion by rat submandibular glands.

We examined the secretory effects of two beta 2-adrenergic agonists, procaterole and terbutaline, on the submandibular glands of anesthetized rats. After stimulation with these agents with and without a range of antagonists (non-specific alpha- and beta-adrenergic blockers), submandibular saliva was collected. The flow rate, protein concentration, the electrophoretic patterns, and amino acid composition of saliva were examined. These parameters were compared with their counterparts in saliva stimulated with isoproterenol (IPR), with and without antagonists. Assessed by these criteria, secreted proteins were classified as the alpha- or beta-type. In addition, IPR-stimulated proteins were compared in submandibular saliva of rats chronically treated with IPR or procaterole. Both beta 2-agonists were potent secretagogues for the submandibular glands of rats. All beta-antagonists completely abolished the secretory effects elicited by both beta 2-agonists, with the exceptions of carteolol and propranolol. However, no blocking agent abolished the secretory effects of IPR (60 mg/kg). The types of proteins in all submandibular saliva samples elicited by both beta 2-agonists with and without antagonists were the beta-type. Enlargement of the submandibular glands was not observed in rats subjected to chronic administration of procaterole, nor were abnormal and additional proteins observed, as confirmed by electrophoresis and by the amino acid analyses.

Animals↗

PubMed2005

Estren promotes androgen phenotypes in primary lymphoid organs and submandibular glands.

BACKGROUND: Estrogens and androgens have extensive effects on the immune system, for example they suppress both T and B lymphopoiesis in thymus and bone marrow. Submandibular glands are sexually dimorphic in rodents, resulting in larger granular convoluted tubules in males compared to females. The aim of the present experiments was to investigate the estrogenic and androgenic effects of 4-estren-3alpha,17beta-diol (estren) on thymus, bone marrow and submandibular glands, and compare the effects to those of 17beta-estradiol (E2) and 5alpha-dihydrotestosterone (DHT), respectively. Estrogen receptors (ERs) were blocked by treatment of mice with the ER-antagonist ICI 182,780; also, knock-out mice lacking one or both ERs were used. RESULTS: As expected, the presence of functional ERs was mandatory for all the effects of E2. Similar to DHT-treatment, estren-treatment resulted in decreased thymus weight, as well as decreased frequency of bone marrow B cells. Treatment with estren or DHT also resulted in a shift in submandibular glands towards an androgen phenotype. All the effects of estren and DHT were independent of ERs. CONCLUSION: Our study is the first to show that estren has similar effects as the androgen DHT on lymphopoiesis in thymus and bone marrow, and on submandibular glands, and that these effects are independent of estrogen receptors. This supports the hypothesis of estren being able to signal through the androgen receptor.

Animals↗

PubMed1981

Postnatal development of trypsin-like esteroproteases in mouse submandibular gland.

The development of trypsin-like esteroproteases in the submandibular gland of mice was studied, using a newly synthesized naphthyl ester (tosyl-L-lysine alpha-naphthyl ester) for the preparation of zymograms and for histochemical demonstration of the enzyme. Esteroprotease activities were first detected spectrophotometrically on day 15 after birth; then increased markedly after day 20. A sex difference in esteroprotease activity appeared on day 25. Zymograms prepared after isoelectric focusing in polyacrylamide slab gels showed that the glands of neonatal mice contained esteroproteases with a rather different composition from that of adult mice. The adult type isozymes appeared first on day 15, and their activities increased markedly after day 20. Histochemical studies revealed that the isozymes of neonatal mice were derived from mast cells. A few striated ducts were first stained on day 15, and the sex difference of the granular tubules became obvious on day 25. These results indicate that the development of trypsin-like esteroproteases faithfully reflects the differentiation of granular tubules in the mouse submandibular gland, except in the neonatal period.

Aging↗

PubMed1988

The short-term effects of a single injection of isoproterenol on proliferation in the submandibular gland, parotid gland and oesophagus in vivo.

Mitotic and labelling indices were studied in the submandibular, parotid and oesophageal cells of male mice within the first 6 hr (but particularly within the 1st hr) of a single injection of isoproterenol or saline, using the metaphase arrest agent (vincristine) which was previously tested for efficacy in submandibular gland. There was a significant increase in the metaphase index of the salivary glands over control values 5, 15, 30, 45 and 60 min after isoproterenol. In contrast, there were no significant changes in the metaphase index of basal cells of the oesophagus. There was no significant change in the labelling index in isoproterenol-treated mice in comparison with saline-injected control animals. Possible explanations for the rapid mitotic response in murine salivary glands are considered; a rapid efflux from G2 into mitosis is thought to be the most likely.

Animals↗

PubMed1978

A stage-restricted secretory system in the submandibular gland of the neonatal rat.

Ribonuclease and amylase have been shown to undergo a transient increase in activity in the submandibular gland of the perinatal rat [5]. We report here that brief stimulation with isoproterenol in vitro selectively releases these enzymes. In addition, analysis by polyacrylamide gel electrophoresis showed that the 4-day old submandibular gland contains three other major protein species that are not present in the sublingual gland and two of which are not evident in the adult submandibular gland. These are also selectively released by the drug. Examination of the glands by light and electron microscopy showed that concurrent with release of these protein products, extensive degranulation occurs in the immature acini (terminal tubules). Our experiments suggest that the 4-day glands show a marked incorporation of 3H-leucine into the submandibular-specific, secreted protein species. This indicates that these proteins will provide convenient molecualr markers of early submandibular differentiation.

Amylases↗

PubMed1985

Effect of chronic treatment with cystic fibrosis fibroblast medium on rat submandibular gland acinar cells.

The effect of chronic treatment with cystic fibrosis (CF) fibroblast medium on rat submandibular gland and pancreas was investigated. Rats were injected for 8 days with conditioned medium from normal or CF fibroblasts. The elemental content of the acinar cells was measured by X-ray microanalysis of cryosections. A significant increase in cellular calcium, and a decrease in cellular sodium concentrations were found after treatment with CF medium. The ultrastructure of the submandibular acinar cells was not affected by the conditioned CF fibroblast culture medium. No effect of treatment with CF medium on ultrastructure and elemental content of pancreatic acinar cells could be demonstrated. The response to alpha-adrenergic, beta-adrenergic, cholinergic, and peptidergic stimulation in submandibular gland acinar cells of rats injected with normal or CF medium was investigated in vitro. With regard to changes in elemental composition after stimulation, no significant differences in response between the two groups could be found. Apparently, a factor in conditioned medium from cultured CF fibroblasts induces a net increase in calcium content of rat submandibular gland acinar cells. Possibly, this factor acts in a similar way in CF patients and may cause elevated calcium levels in CF cells.

Adolescent↗

PubMed1977

[The ciliated epithelium in the efferent ducts of the submandibular gland].

The examination of 442 bioptic and necroptic submandibular salivary glands revealed that in cases of chronic inflamation of the gland (125 cases) the epithelial lining of its ducts contained ciliated colummar epithelium in 57% of cases. The finding of ciliated epithelium was associated with lithiasis of the gland in nearly 79% of cases. Normal submandibular glands or glands affected with pathological conditions other than chronic inflammation did not contain ciliated epithelium. The present paper is the first one in the literature purposedly dealing with the presence of ciliated epithelium within the ducts of the submandibular gland.

Epithelium↗

PubMed1987

Isolation, characterization, and localization of antigen gamma, a serine proteinase of the "kallikrein-family" in the rat submandibular gland.

A trypsin-like serine proteinase, antigen gamma, immunologically partially identical to glandular kallikrein when run against anti-rat glandular kallikrein antiserum in immunoelectrophoresis, was purified from the rat submandibular gland. The enzyme was purified by a two-step chromatography procedure, ionexchange chromatography followed by gel filtration. The criteria for purity were one band in SDS-polyacrylamide gel electrophoresis and in immunoelectrophoresis, respectively. Antigen gamma had a molecular mass of 25,000 Da and consisted of two polypeptide chains with molecular masses of 14,000 and 11,000 Da. The preparation contained several isoenzymes with pI ranging from 4.1 to 4.5. The enzyme showed high specific enzyme activity against the substrate D-valyl-L-leucyl-L-arginine-4-nitroanilide (S-2266), some trypsin-like and kininogenase activity, but no angiotensin converting enzyme, kininase, or tonin activity. Amidolytic activity was increased and stabilized by the presence of detergent in the assay buffer. The pH-optimum of antigen gamma amidolytic activity was about 10. Antigen gamma was inhibited by SBTI and PMSF, whereas aprotinin had to be added in a more than 100 times higher concentration than for glandular kallikrein. The binding pattern of antigen gamma to plasma proteins was different from that of tonin and glandular kallikrein. Antiserum against antigen gamma was raised in rabbits and characterized against rat submandibular gland homogenate. Immunohistochemistry showed antigen gamma in the secretory granules of the submandibular gland granular tubular cells but only adhering to the luminal cell wall in the striated and main excretory ducts. Antigen gamma was not detected in the sublingual or parotid gland or in the kidney. Antigen gamma was demonstrated by immunoelectrophoresis in rat submandibular gland saliva. The concentration was higher in sympathetically than in parasympathetically induced secretion.

Angiotensin-Converting Enzyme Inhibitors↗

PubMed1984

Calcium-dependent enhancement by carbachol of the VIP-induced cyclic AMP accumulation in cat submandibular gland.

The interaction of two coexisting transmitters in the cat submandibular gland has been elucidated by studying effects of VIP and carbachol on cyclic AMP accumulation in isolated acini from the gland. Carbachol was found to potentiate the cyclic AMP increase induced by VIP by an atropine sensitive mechanism. The effect of carbachol on cyclic AMP accumulation was abolished by including EGTA in the incubation medium as was the carbachol mediated potentiation of VIP responses. The calmodulin inhibitor trifluoperazine had a similar, but less marked effect. The effect of carbachol was mimicked by phenylephrine (30 microM) and by the calcium inophore A 23187 (3 microM), and also by ethanol in a concentration reported to enhance membrane fluidity. The phospholipase A2 inhibitor, mepacrine, tended to decrease carbachol actions. Our results show that the potentiation of VIP responses in feline submandibular gland is calcium-dependent. The mechanism could involve a calcium-calmodulin-induced stimulation of adenylate cyclase or calcium-induced change in membrane phospholipid metabolism.

Animals↗