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PubMed2004

Molecular cloning and characterization of the neonatal rat and mouse submandibular gland protein SMGC.

We report the molecular cloning and characterization of SMGC, a major secretory product and a marker of the type I (terminal tubule) cells of the neonatal rat and mouse submandibular gland. SMGC is expressed in the submandibular gland at high levels through postnatal day 20, but in the adult is present only in some intercalated duct cells. Rat and mouse SMGC have deduced molecular weights of 67.8 and 74.4 kDa, respectively, are 37% Ser+Gly+Thr, and contain tandem repeats of between 8 and 60 amino acids. Secreted SMGC visualized by SDS-PAGE and silver staining is 89 kDa in rat and 105 kDa in mouse, although Western blot analyses with anti-SMGC antisera demonstrate multiple additional lower molecular weight forms. Contributions to the heterogeneity of SMGC include alternate splicing, proteolysis and N-glycosylation. Smgc is localized on rat chromosome 7q34-35 and on mouse chromosome 15E3, both immediately upstream of the high molecular weight salivary mucin, Muc19. Amino acid sequence identity between the signal peptides of SMGC, human MUC19 and pig submaxillary mucin suggest that rat and mouse Smgc and Muc19 arose from a single ancestral mucin gene.

Alternative Splicing↗
PubMed1987

Characterization of the vasoactive intestinal peptide receptor in rat submandibular gland: radioligand binding assay in membrane preparations.

The vasoactive intestinal peptide (VIP) receptor in membranes from rat submandibular gland was studied using radioligand binding assays with 125I-VIP and various unlabeled competing ligands. In addition to the necessity of working within the parameters under which all radioligand binding assays should be performed, binding studies with 125I-VIP, as with other peptide hormones and neurotransmitters, are subject to additional technical difficulties. Specific problems that were addressed included radioligand proteolysis, the identification of an effective protease inhibitor (leupeptin) and the deleterious effects of a commonly used inhibitor (bacitracin); avid radioligand absorption to incubation tubes that was eliminated by precoating of the tubes with a combination of polyethylenimine and an organosilane; and a disproportionate effect of increasing membrane protein concentration on affinity estimates. Under optimized conditions, the affinity (Kd) and density Bmax values for 125I-VIP obtained from saturation assays (76 pM, 2.0 pmol/mg) were in excellent agreement. Membrane protein (or receptor) levels beyond the linear portion of the receptor concentration curve are often used in radioligand binding assays. Results from 125I-VIP binding studies at elevated receptor concentrations revealed the predicted marked decrease in receptor affinity. In addition, the rank order potency of unlabeled ligands in inhibition binding assays was changed. The optimization of the assay for measuring VIP receptors in submandibular gland membrane provides a reliable method for studying the role of receptor regulation in stimulus-secretion coupling for this neuropeptide.

Animals↗
PubMed1983

Immunohistochemistry of tonin in rat submandibular gland during development, lactation and secretion.

Tonin is a serine protease found in high concentrations in the submandibular gland (SMG) of the adult rat where it has been localized by immunohistochemistry in the granular ducts. The present study examined the development of tonin in the SMG, the effect of lactation and of stimulation of tonin release, using the peroxidase-antiperoxidase technique and antitonin. Tonin-like immunoreactivity first appeared in the primitive striated duct of the SMG on day 19 foetal and increased in intensity as the ducts developed into granular ducts. Reaction product in granules was seen on day 17 postpartum. Its localization within granules was established by immunochemistry of Sepharose beads to which had been coupled the contents of granules isolated from adult rats. The granular ducts of female rats, which are less developed than in the male, showed a marked increase in tonin-immunoreactivity during lactation. Stimulation of tonin secretion by isoprenaline caused massive discharge of tonin-like immunoreactivity into the lumen of the granular ducts during in vitro incubation. However, within one hour complete regranulation was apparent. The secretion was prevented by propranolol. The results indicate that tonin or a tonin-like substance appears in the rat submandibular gland late in gestation in ducts that presumably develop into granular ducts where it is found in abundance in granules in the adult, that the amount in females is increased during lactation, and that most of the granules are discharged during stimulation, only to be rapidly replaced.

Animals↗
PubMed1985

The chronically reserpinized rat: decreased glycolytic activity in the submandibular gland.

Some of the enzymes and metabolites of the glycolytic pathway of an animal model for cystic fibrosis (the chronically reserpine-treated rat) were investigated. The activities of the enzymes phosphofructokinase (P less than 0.002), enolase (P less than 0.03), pyruvate kinase (P less than 0.005), and lactate dehydrogenase (P less than 0.009) were decreased whereas the activity of glycerol-3-phosphate dehydrogenase was unaffected in the submandibular glands of the treated animals. For metabolites, the reserpine treatment resulted in an increased concentration of glycogen (P less than 0.0002) and phosphoenolpyruvate (P less than 0.001) and a decreased concentration of pyruvate (P less than 0.005) and lactate (P less than 0.002) in the glands. The concentration of glucose and glycerate-2-phosphate was unaffected. The perchloric acid-soluble part of the proteins was also increased (P less than 0.0001) in the submandibular glands of the reserpine-treated animals, as was the activity of ribonuclease. These findings point to a disturbance in the metabolism of glucose and a possible acidosis in the submandibular glands of this animal model for cystic fibrosis.

Animals↗
PubMed1975

Metastatic malignancy in the submandibular gland.

Metastatic disease in the major salivary glands has been observed and reported in the literature. Often the site of origin of the primary tumor is the skin or mucosal lining of the head and neck structures, and the primary tumors most commonly are melanomas or squamous-cell carcinomas. The most frequently involved salivary gland is the parotid. The submandibular and sublingual glands are rarely involved by mestastatic disease. Secondary lesions arising from distant primary tumors may occasionally involve the parotid gland, but they are extremely rare in the other major salivary glands. The second reported case of metastatic carcinoma in the submandibular gland is presented here. Our case and the previously reported case are similar in that both primary tumors originated in the breast. The clinical and morphologic features are reviewed.

Adenocarcinoma↗
PubMed1988

Inhibitory effects of atropine and adrenergic antagonists on the changes in autonomic receptors and cyclic nucleotides of rat parotid and submandibular glands caused by sympathetic nerve stimulation.

The changes in cyclic AMP (cAMP) concentration and density of beta-adrenoceptors caused by electrical stimulation of the sympathetic innervation to parotid and submandibular glands of rat did not occur when the alpha- and beta-adrenergic antagonists, phentolamine, and propranolol were administered 20 min prior to initiation of stimulation. They also did not occur when phentolamine, the beta-adrenergic antagonist, was administered alone prior to nerve stimulation, indicating that beta-adrenoceptors mediate these effects. Simultaneous administration of the alpha- and beta-antagonists also prevented the changes in densities of muscarinic receptors and cGMP concentrations usually induced by sympathetic nerve stimulation. Also, the changes in muscarinic receptors and cGMP did not occur when atropine was administered prior to nerve stimulation, nor did they occur with simultaneous administration of atropine, phentolamine + propranolol; with phentolamine alone, or propranolol alone, the effects were blocked to a large extent. Secretion was inhibited completely when both adrenergic antagonists were present during nerve stimulation, but flow rate was unchanged when atropine was present. The changes in both beta-adrenoceptors and muscarinic receptors reflect a desensitization caused by prolonged exposure to neurotransmitters released when the sympathetic nerve is stimulated. The changes are prevented when either atropine or adrenergic antagonists are present during nerve stimulation.

Adrenergic beta-Antagonists↗
PubMed1984

Submandibular glands as organs of synthesis and accumulation of insulin-like protein.

Experiments are presented to determine the endocrine function of submandibular salivary glands in the light of a hypothesis about synthesis of insulin-like protein (ILP) and participation of these organs in regulation of the glucose level in the organism. ILP was discovered in submandibular glands of male mice, rats, hamsters, cats, rabbits, pigs, bulls, and men. Electrophoretical and immunological studies of insulin-like protein, extracted from these glands, showed its similarity with pancreatic insulin. Immunofluorescent studies showed that ILP was localized in the granular duct cells of mice and rats. Studies by immunoperoxidase method illustrated that insulin-like protein was distributed along the periphery of the electron-dense secretory granules. In submandibular glands of animals which do not contain granular ducts, ILP was revealed in the striated tubules (by immunofluorescent method). Study of submandibular glands in vitro showed that ILP was synthesized in glands and not simply accumulated there. Under the condition of insulin deficiency (in vivo) and increased glucose concentration in culture media (in vitro), duct cells of submandibular glands released ILP in increasing amounts. Injection of ILP into mice caused a decrease of glucose concentration in the blood and a convulsive reaction. This fact indicates the similarity of ILP with insulin in its physiological action. Our studies permit to draw the conclusion on the participation of submandibular glands in the regulation of glucose homeostasis.

Animals↗
PubMed2001

The case of the submandibular gland sialolith.

The authors describe a case of a sialolith formation in a submandibular gland over 2 years. A large salivary stone is demonstrated radiologically and surgical treatment is described.

Humans↗
PubMed1987

Does calmodulin participate as an intermediate in the process of glycogenolysis in rat submandibular gland?

A role of calmodulin in the process of the glycogenolysis induced by adrenaline was investigated in rat submandibular gland. When the slice was incubated in the Ca2+-free medium, the adrenaline-induced glycogenolysis was significantly inhibited. Moreover, the inhibition of glycogenolysis induced in the Ca2+-free medium was further enhanced by addition of 2 mM EGTA. Chlorpromazine, trifluoperazine and W-7, calmodulin antagonists, had no effect on the glycogenolysis induced by adrenaline. These results suggest that the adrenaline-induced glycogenolysis is dependent on the presence of Ca2+ and that calmodulin does not participate in the process of glycogenolysis in rat submandibular gland.

Animals↗
PubMed2002

Differences in the regulatory mechanism of amylase release by rat parotid and submandibular glands.

It is not known whether the mechanisms involved in amylase release in submandibular and parotid glands are similar. Here, the participation of different signalling pathways in amylase release by the parotid and submandibular glands of the male rat was compared by studying the secretory response after beta-adrenergic stimulation. The beta-adrenergic agonist isoproterenol induced an increase of cAMP in both salivary glands, but while in the parotid it triggered amylase release, in the submandibular it was unable to increase amylase secretion. Parotid amylase release was dependent on adenylate cyclase activation, as SQ-22536 inhibited the secretory effect. In contrast, submandibular amylase secretion did not depend on the intracellular concentration of cAMP, as SQ-22536 did not modify its secretory response. Moreover, other activators of adenylate cyclase, such as forskolin and prostaglandin E2, also failed to modify amylase release by the submandibular gland. Neither ionophores nor calcium-blocking agents, as well as calcium-calmodulin and nitric oxide synthase inhibitors, were effective in modifying basal amylase release by the submandibular gland. However, the disruption of microfilaments with cytochalasin B, but not the disruption of microtubules with colchicine, prevented amylase release in that gland. It is concluded that amylase exocytosis in the submandibular gland is a constitutive non-regulated phenomenon, as it is independent of extracellular or intracellular signals. It depends only on the integrity of the microfilaments, probably used by the vesicles to travel from the Golgi apparatus to the plasma membrane.

Actin Cytoskeleton↗
PubMed1979

[The buccal approach for excision of posterior submandibular gland calculi. A report on 10 cases (author's transl)].

The authors describe ten patients with calculi in various regions of the submandibular gland, who were operated upon using the buccal approach, and review the different therapeutic procedures available. Though removal of the submandibular gland is a mutilating operation and has many sequelae when operated upon through the submandibular approach, it is still, surprisingly, the standard operative procedure for removal of posterior calculi, even though Delaire had already expressed his logical preference for the buccal approach, 23 years ago. This procedure was used in the 10 cases reported here and results were very good, and there were no sequelae. A strict operative technique is necessary, including general anaesthesia and dissection of Wharthon's duct along its whole length as the essential factors.

Adult↗
PubMed1983

Effect of indomethacin on the blood flow of the rabbit submandibular gland.

Effects of the stimulation of the chorda tympani and acetylcholine infused into the artery of the gland were studied on the blood flow and saliva secretion of the rabbit submandibular gland before and after indomethacin treatment. The enhancement of blood flow and saliva secretion was less apparent after indomethacin than before. Results suggest that endogenous prostaglandins are also involved in the mechanism of the augmentation of blood flow and saliva secretion produced by the stimulation of the chorda tympani in the rabbit submandibular gland.

Acetylcholine↗
PubMed1981

Cyclic AMP increase by histamine and its analogues in guinea-pig submandibular gland.

The effect of histamine and its analogues on the cyclic AMP level in chopped guinea-pig submandibular glands was investigated. These agonists increased the cyclic AMP level dose-dependently. The ED50 values for histamine and 4-methylhistamine were approximately 1.3 x 10(-5) M and 2.2 x 10(-5) M, respectively. The efficacy of 4-methylhistamine was higher than that of histamine. 2-(2-Pyridyl)-ethylamine was far less effective compared with histamine or 4-methylhistamine. Metiamide markedly antagonized the cyclic AMP response to histamine and 4-methylhistamine, but mepyramine was ineffective. A combination of aminoguanidine and quinacrine had no influence on the effect of histamine on the cyclic AMP level. These results indicate the H2-type histamine receptors which mediate the cyclic AMP increase are present in the guinea-pig submandibular gland.

Animals↗
PubMed1993

Effects of benzo(a)pyrene and nicotine on prostaglandin synthesis in buccal pouch and submandibular glands of the Syrian hamster.

Adult male Syrian hamsters were treated by swabbing the apex of the buccal pouch with corn oil (control, C), 1 mM benzo(a)pyrene (BP), nicotine (NC), or BP+NC in corn oil, twice daily, 5 days a week. After a 4-week treatment, the pouches and submandibular glands were dissected and used for the determination of endogenous prostaglandin (PG) production and studies on in vitro PG synthesis. Of the three PGs analysed (PGE2, PGF2 alpha and 6-keto-PGF1 alpha), PGE2 was predominant in the pouch and the glands. BP or NC alone had only a weak effect on PG synthesis in both tissues. However, the combination of BP and NC had a synergistic effect, causing diminished PG synthesis in both tissues. In buccal pouch, BP+NC significantly decreased the concentrations of endogenous PGE2 and PGF2 alpha (PGE2: 0.669 +/- 0.254 versus 1.698 +/- 0.460, PGF2 alpha: 0.273 +/- 0.090 versus 0.625 +/- 0.272 ng/g tissue; BP+NC versus C; mean +/- SD, n = 5, p < 0.05). Similarly significant results were also found for in vitro PG synthesis (PGE2: 0.541 +/- 0.249 versus 1.399 +/- 0.340, PGF2 alpha: 1.045 +/- 0.428 versus 2.133 +/- 0.510 ng/g tissue; BP+NC versus C; mean +/- SD, n = 5, p < 0.05). In submandibular glands, BP+NC significantly diminished the concentration of endogenous PGE2 (1.183 +/- 0.175 versus 2.379 +/- 0.488 ng/g tissue; BP+NC versus C; mean +/- SD, n = 5, p < 0.05). The synthesis of 6-keto-PGF1 alpha in both tissues, and the synthesis of PGF2 alpha in submandibular gland, were slightly decreased with all treatments.

6-Ketoprostaglandin F1 alpha↗
PubMed1984

Developmental changes of trypsin-like protease activity in the submandibular glands of male and female young growing rats.

The changes during infancy of trypsin-like protease activity in the submandibular glands of male and female rats (4-20 weeks of age) were observed. Activity was low at 4 weeks of age, then increased markedly at 8-10 weeks of age. Seven peaks (I-VII) with isoelectric points of 4.0, 4.3, 4.5, 5.0, 5.4, 5.7 and 6.4 of protease activity were observed in the glands of adult rats by isoelectric focusing. Only peaks I and II were detected in the glands at 4 weeks of age in both sexes. After 8 weeks, all 7 peaks appeared in the glands of males and females. Repeated injection of isoprenaline into rats of 7 weeks of age elevated markedly the weight of submandibular glands; the elevation of protease activity and of inhibitory activity against dopa-decarboxylase was not found. These results indicate that proteases in peaks I and II appear at an age close to that of development of acini; protease activity in peaks III-VII increases is close to the time of development of the granular ducts.

Animals↗
PubMed1998

Inhibitory effects by a submandibular gland extract on luteinizing hormone-stimulated testosterone production by testicular cells.

In order to evaluate the role of the submandibular gland (SMG) on testosterone (T) production by the testis, primary cultured testicular cells were prepared from rats that had the submandibular gland surgically ablated (G-) and control (sham operated) rats (S.O.) respectively. The cells were incubated with or without 100 ng/ml luteinizing hormone (LH) and/or SMG extract. The same linear increase in T secretion was shown by both S.O. and G- cells on multi-stimulation with LH for up to 96 hrs. However, while an equivalent response was shown for S.O. cells after a single LH stimulation at 96 hrs, T secretion by the G- cells reached a plateau after 24 hrs. The level at 96 hrs was thus approximate 30% and 33% of those of S.O. cells with and without multi-stimulus by LH for 96 hrs, respectively. When S.O. cells were cultured with SMG extract, LH-stimulated T secretion was dose-dependently inhibited and there was no effect on basal T secretion. The inhibitory effect was abolished by treatment at 95 degrees C for 5 min. Ultra-filtration indicated that the molecular size of the inhibitory agent was greater than 30,000. It is proposed that SMG may contain a high molecular weight, heat labile soluble factor(s) which affects T secretion by inhibiting LH action in testicular cells.

Animals↗
PubMed1988

Transient involvement of enkephalins in both the sympathetic and parasympathetic innervations of the submandibular gland of rats. Light- and electron-microscopic immunocytochemical study.

A time course study with enkephalin (Enk)-like immunoreactivity has revealed that nerve fibers intensely immunoreactive for Enk-8 appeared transiently only during the postnatal week 2 and 4 within the acini as well as in the inter- and intralobular connective tissues of the submandibular gland of rats. At these stages numerous nerve fibers immunoreactive for tyrosine hydroxylase (TH) appeared also in the inter- and intralobular connective tissues and within the acini. Coincidently with these postnatal stages, abundant Enk-immunoreactive principal ganglion cells appeared in the superior cervical ganglion. These were not immunoreactive for neuropeptide tyrosine (NPY). A substantial number of Enk-immunoreactive ganglion cells were also present in the submandibular ganglia at these younger postnatal stages. Superior cervical ganglionectomy at these stages resulted in a marked decrease in number of the inter- and intralobular Enk-immunoreactive nerve fibers, a slight decrease in number of the intraacinar Enk-immunoreactive nerve fibers, and almost complete disappearance of intraglandular TH-immunoreactive nerve fibers. Immuno-electron-microscopic analysis revealed that Enk-immunoreactive nerve fibers in the submandibular gland were identified as electron-dense neuronal profiles enclosed by Schwann cells in the inter- and intralobular connective tissues and those directly apposed to secretory cells within the acini. They contained small clear vesicles mixed with some large granular vesicles. After postnatal week 6, no Enk-immunoreactive nerve fibers were detected in the submandibular gland, and no TH-immunoreactive nerve fibers were seen within the acini, while TH-immunoreactive nerve fibers remained numerous in the inter- and intralobular connective tissues. These findings indicate that both sympathetic and parasympathetic nerve fibers exhibit Enk-like immunoreactivity transiently during postnatal weeks 2 and 4.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1987

The application of microspectrocytofluorometric measurement of Feulgen nuclear DNA content to experimental tumors of rat submandibular gland. 1. Pathogensis and nuclear DNA content.

Pellets containing 9,10-dimethyl-1,2-benzanthracene (DMBA) were inserted into the submandibular glands of 65 male Fischer rats to determine the nuclear DNA content of both metaplastic epithelium during the process of development of squamous metaplasia (noted at 1, 2, 5, 7, 9, 10, 11, 13 and 15 weeks after insertion) and tumor cells of squamous cell carcinoma 24 weeks after insertion. From the second week of insertion onward, squamous metaplasia was found in excretory duct remaining epithelium in the tissues around the pellet. Dysplasia or a lining of metaplastic epithelium with atypia adjacent to pellets (i.e., formation of an epidermal cyst) was observed 5 weeks after insertion. Coincident with this dysplasia or metaplastic epithelium with atypia, variation in the nuclear DNA content was observed, showing the emergence of octaploids and a widespread histogram pattern with many peaks. Between 7 and 15 weeks, 25%-66% of the lining metaplastic epithelium showed variation in the nuclear DNA content due to shift of a peak (major mode) to a triploid as well as a vague major mode with the appearance of octaploids, etc. Tumors of the submandibular gland were recognized at 24 weeks in 11 (55%) of 20 rats. Nine were keratinized squamous cell carcinomas and the remaining 2 were carcinosarcomas. Marked abnormalities in the nuclear DNA content were observed in 5 of the 11 rats (45.5%), 3 with keratinizing squamous cell carcinoma and 2 with carcinosarcomas. In contrast, no marked variation in the nuclear DNA content was found in 5 rats in which the tumor was associated with cysts.

9,10-Dimethyl-1,2-benzanthracene↗