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PubMed1987

Isolation of a thiol-activated T-kininogenase from the rat submandibular gland.

T-kininogenase (T-kgnase) activity has been investigated in tissues of the rat and submandibular glands of the rat, mouse and guinea pig. Both rat and mouse submandibular homogenates showed high T-kgnase activity. The enzyme has been purified 360-fold from rat submandibular gland homogenate supernatant fluid. The enzyme has an apparent molecular mass of 28 kDa and a pH optimum of 8.0 toward T-kininogen. It cleaved T-kininogen in catalytic quantities to release T-kinin (Ile-Ser-bradykinin) and small quantities of bradykinin and an unknown kinin. The activity of the enzyme was increased 10-fold in the presence of thiol groups (dithiothreitol) and inhibited by leupeptin (90%) and to a lesser extent by aprotinin (49%), TLCK (46%) and soybean trypsin inhibitor (27%). Pepstatin and PMSF did not inhibit the enzyme. Studies on substrate specificity, pH optimum and agents which inhibit T-kgnase activity demonstrate that this enzyme is different from plasma and tissue kallikreins, cathepsin D, esterase A and esterase B (other known kininogenases). It is the first thiol-activated kininogenase to be reported.

Animals↗
PubMed1985

Binding of glucocorticoid to the androgen receptor of mouse submandibular glands.

An increase in esteroprotease activity, a known cytodifferentiating response to androgen in the submandibular gland, occurred after cortisol acetate, dexamethasone or methyltrienolone (R1881) treatment in castrated genetically normal male (X/Y-castrated) mice, but not in normal male (X/Y) and testicular feminized male (Tfm/Y) mice. A peak with specific binding for [3H]cortisol appeared in sucrose density gradient patterns of extracts from X/Y-castrated mice and in almost the same fraction number as that for [3H]R1881 binding. However, peaks specific for neither [3H]cortisol nor [3H]R1881 binding were observed in Tfm/Y mice. The peak binding [3H]cortisol in extracts from X/Y-castrated mice, as well as the one binding [3H]R1881, were inhibited by unlabelled R1881 and cyproterone acetate, an antiandrogen; the peak was not, however, affected by unlabelled oestradiol-17 beta. The binding capacity of [3H]cortisol determined by Scatchard analysis was similar to that of [3H]R1881 (103 and 106 fmol/mg protein respectively). The Kd value of [3H]cortisol, however, was about 13.6-fold higher than that of R1881. These results suggest that cortisol has the ability to promote androgenic cytodifferentiating action in the mouse submandibular gland by binding to its androgen receptors, if androgens are absent or deficient.

Animals↗
PubMed1993

Histochemical localization and X-ray microanalysis of calcium in the rat submandibular gland: demonstration of possible sites for microlith induction.

Rapidly frozen and freeze-substituted submandibular glands of young female rats were embedded in Epon and processed for histochemical demonstration of calcium with the glyoxal bis (2-hydroxyanil) (GBHA) staining method. GBHA staining of thick Epon sections revealed discrete calcium reactions of moderate intensity in practically every secretory granule but not in other compartments of the acinar cells. The saliva in the excretory duct was also reactive with GBHA and showed a drastic decrease in staining intensity toward the distal segments of excretory ducts with larger diameters. In addition, the duct saliva contained numerous tiny particles that were highly GBHA reactive. Stromal cells and cells lining the excretory duct were totally free of reactions. In the acinar cells, X-ray analysis detected distinct peaks for calcium in secretory granules and smaller ones in the Golgi apparatus, while they were undetectable in the rough surfaced endoplasmic reticulum (RER), implicating post-RER calcium loading in the secretory pathway. Electron-dense deposits in the duct saliva showed distinct peaks both for calcium and phosphorus, though these appeared in the acinar secretory granules and other cytoplasmic regions lacked phosphorus. Our observations thus demonstrated physiological calcium in the intra- as well as extracellular compartments of the submandibular gland, and further confirmed drastic changes in chemical composition along the synthetic and secretory pathways of the saliva, by both histochemical and X-ray microanalytical methods. GBHA staining of calcium combined with X-ray microanalysis is useful for an evaluation of the physiology and histo-pathological changes of the salivary glands associated with initial phases of microliths as well as sialoliths formation.

Aminophenols↗
PubMed1994

Effects of cholinergic and adrenergic agonists on the secretion of fluid and protein by submandibular glands of the guinea-pig and the mouse.

Significant differences were observed between the guinea-pig and the mouse in terms of the secretion of fluid, protein and secretory granules from submandibular glands in response to pilocarpine, phenylephrine and isoproterenol. In both the guinea-pig and the mouse, the secretory responses induced by pilocarpine, phenylephrine and isoproterenol were inhibited by pretreatment with 4-DAMP, phentolamine and propranolol, respectively. The results suggest that the submandibular glands of the guinea-pig and the mouse have M3-cholinoreceptors, as well as alpha- and beta-adrenoceptors, and that these receptors play different roles in the secretion of fluid, protein and secretory granules from guinea-pig and mouse submandibular glands.

Adrenergic Agonists↗
PubMed1994

Basolateral K+ efflux is largely independent of maxi-K+ channels in rat submandibular glands during secretion.

The involvement of large-conductance, voltage- and Ca(2+)-activated K+ channels (maxi-K+ channels) in basolateral Ca(2+)-dependent K(+)-efflux pathways and fluid secretion by the rat submandibular gland was investigated. Basolateral K+ efflux was monitored by measuring the change in K+ concentration in the perfusate collected from the vein of the isolated, perfused rat submandibular gland every 30 s. Under conditions in which the Na+/K(+)-ATPase and Na(+)-K(+)-2Cl- cotransporter were inhibited by ouabain (1 mmol/l) and bumetanide (50 mumol/l) respectively, continuous stimulation with acetylcholine (ACh) (1 mumol/l) caused a transient large net K+ efflux, followed by a smaller K+ efflux, which gradually returned to the basal level within 10 min. These two components of the K+ efflux appear to be dependent on an increase in cytosolic Ca2+ concentration. The initial transient K+ efflux was not affected by charybdotoxin (100 nmol/l) or tetraethylammonium (TEA) (5 mmol/l) but the smaller second component was strongly and reversibly inhibited by charybdotoxin (100 nmol/l) and TEA (0.1 and 5 mmol/l). The initial K+ efflux transient induced by ACh was inhibited by quinine (0.1-3 mmol/l), quinidine (1-3 mmol/l) and Ba2+ (5 mmol/l), but not by verapamil (0.1 mmol/l), lidocaine (1 mmol/l), 4-aminopyridine (1 mmol/l) or apamin (1 mumol/l). Ca(2+)-dependent transient large K+ effluxes induced by substance P (0.01 mumol/l) and A23187 (3 mumol/l) were not inhibited by TEA (5 mmol/l or 10 mmol/l). A23187 (3 mumol/l) evoked a biphasic fluid-secretory response, which was not inhibited by TEA (5 mmol/l). Patch-clamp studies confirmed that the whole-cell outward K+ current attributable to maxi-K+ channels obtained from rat submandibular endpiece cells was strongly inhibited by the addition of TEA (1-10 mmol/l) to the bath. It is concluded that maxi-K+ channels are not responsible for the major part of the Ca(2+)-dependent basolateral K+ efflux and fluid secretion by the rat submandibular gland.

Acetylcholine↗
PubMed1992

Immunohistochemical localization of rat submandibular gland esterase B (homologous to the RSKG-7 kallikrein gene) in relation to other serine proteases of the kallikrein family.

The rat submandibular gland contains several members of the kallikrein family. In the present study we purified and raised an antiserum against one of these enzymes, i.e., esterase B, which was first described by Khullar et al. in 1986. N-terminal amino acid analysis revealed complete homology between esterase B and the kallikrein family gene RSKG-7. For characterization of the antiserum, flat-bed isoelectrofocusing with immunoblotting was superior to immunoelectrophoresis and double immunodiffusion in detecting and identifying crossreacting proteins. This was due to the fact that kallikrein-like enzymes were readily separated by isoelectrofocusing, and immunoreactivity was easily detected by the sensitive peroxidase-anti-peroxidase staining after blotting onto nitrocellulose membrane. Immunohistochemical controls were carried out accordingly, including homologous as well as crossreacting antigens. In the submandibular gland, esterase B was detected exclusively in all granular convoluted tubular cells, co-localized with tissue kallikrein and tonin. Some staining was also observed in striated duct cells; however, this staining reaction was induced by cross-reactivity with kallikrein, since staining was abolished by addition of kallikrein as well as esterase B to the primary antiserum. It was therefore concluded that like tonin and antigen gamma, but unlike kallikrein, esterase B was not detected in the striated ducts of the submandibular, parotid, or sublingual glands. This separation in anatomic distribution between esterase B and kallikrein may indicate that prokallikrein activation is not the only biological function of esterase B.

Amino Acid Sequence↗
PubMed2005

Valproic acid-associated sialadenosis of the parotid and submandibular glands: diagnostic and therapeutic aspects.

Sialadenosis has been defined as a non-inflammatory, parenchymatous salivary gland disease causing recurrent, bilateral swelling of the salivary glands. As an adverse drug reaction of valproic acid, sialadenosis is very rare. To our knowledge, it has been reported only once in the world literature to date. We present herein the case of a patient with valproic acid-associated sialadenosis of both the parotid and submandibular glands. This appears to be the first published case of a patient who received surgical treatment. On light and electron microscopy of all the affected salivary glands, granular sialadenosis with predominantly moderate electron-dense secretory cytoplasmatic granules was observed. No relevant degenerative alterations were seen. There was no histological evidence of peripheral neuropathy of the nerve supply, leading to disordered activity of acinar cells by loss of neurosecretory granules. Lateral parotidectomy, performed under neuromonitoring control for safety reasons, is the treatment of choice for chronic recurrent parotitis that does not respond to conservative therapy, particularly if the cosmetic deformity is unacceptable to the patient. If the submandibular glands are involved, partial removal is recommended.

Adult↗
PubMed1978

Developmental distribution of microperoxisomes in the rat submandibular gland.

The present study investigated the size, number, and distribution of microperoxisomes (MP) during the prenatal and postnatal development of the rat submandibular gland (SMG). A three-fold increase in MP number per cell was observed in the cells of the rudiment from the 15th to the 16th day of gestation. The early secretory and striated duct cells contained about 9.0 MP. The number of MP per secretory cell decreased such that 3.5 MP were found in each mature acinar cell. In the striated duct cells, MP number progressively increased to 40.0. As the convoluted granular tubule cells (CGT) developed from striated duct cells there was an increase in MP number from 16.0 to 26.0/cell. At maturity, the convoluted granular tubule cells contained only 14.0 MP. Throughout development of the SMG, intercalated duct cells showed only rare MP. The data suggests that the number, size, and distribution of MP changes as a function of the particular path of differentiation followed by the various cells in the rat SMG.

Animals↗
PubMed1990

Expression of glutamine/glutamic acid-rich proteins in rat submandibular glands.

The expression of GRP transcripts was found to be highly specific to the rat submandibular gland. GRP cross-reactive species were detected in the saliva of both inbred and outbred rat strains. There was no evidence of GRP transcripts in RNA prepared from bovine, ovine, porcine or murine submandibular glands. Thus the GRPs differ from the family of PRPs that are expressed in several species. The restriction of GRP expression to the rat suggests a relatively recent origin for a functional GRP gene, presumably after rat-mouse divergence. The ontogeny of the relative steady-state levels of GRP transcripts was assessed by dot blot analysis. Maximal levels of RNA-encoding GRP were detected at 6 months; there was a significant age-related decline at both 12 and 18 months. There was, however, no significant age-related alteration in the size of transcripts which encode this protein family.

Animals↗
PubMed1992

Differential regulation of agonist-stimulated Ca2+ influx in acini of rat pancreas and submandibular gland.

In order to characterize agonist-dependent Ca2+ influx pathways, changes in cystolic Ca2+ (Ca2+i) during stimulation with multiple Ca(2+)-mobilizing agents were measured in rat pancreatic and submandibular gland acini loaded with fura-2. In pancreatic acini, maximal levels of carbachol and cholecystokinin octapeptide (CCK-8) produced virtually identical changes in Ca2+i when added alone or together: an immediate increase to 4-5 times resting Ca2+i followed by a decline to a steady-state level 2-3 times resting, which was unchanged by another stimulus. In submandibular gland acini, maximal carbachol stimulation increased Ca2+i 3-4-fold followed by a plateau at 2-3 times resting, which was further increased by epinephrine. Epinephrine alone increased steady-state Ca2+i to 53 +/- 18% (n = 21) of that observed with carbachol. Stimulation with both agents increased the steady-state plateau level of Ca2+i to 144 +/- 28% of that during exposure to carbachol alone (n = 11, p < 0.05). When changes in Ca2+i due solely to Ca2+ influx were measured, carbachol and epinephrine together increased Ca2+i during the steady-state phase to 149 +/- 31% of that measured with carbachol alone (n = 8, p < 0.05). Atropine blocked only responses to carbachol, prazosin blocked only responses to epinephrine and L 364,718 blocked only CCK-8-induced changes in Ca2+i. Thus, in pancreatic acini, a single agonist-sensitive Ca2+ influx pathway is linked independently to muscarinic cholinergic and peptidergic (CCK-8) receptors. In contrast, submandibular gland acini contain functionally separate agonist-sensitive Ca2+ influx pathways, which are independently linked to muscarinic and to alpha-1 adrenergic receptors.

Animals↗
PubMed1990

[The effects of solid or liquified diet on the submandibular glands of mice with age].

The influence of properties of the diets on the retardation of growth of the jaw bones and masticatory muscles has been established. As the intake of liquid foodstuffs increases in the human diet, it is considered possible that these liquid diets may retard the growth of human salivary glands. 180 newborn C3H/He strain male mice were divided into solid and liquid diet groups and then the development and aging of the submandibular glands were observed both in function and morphologically until 60 weeks of age. The following results were obtained: 1. No differences could be detected in body length and body weight of the mice between solid and liquid diet groups. The wet weight of the liver in the liquid diet group was significantly larger than the solid diet group, which suggested that the liquid diet may have accelerated metabolic functions. Additionally, masticatory muscles of the liquid diet group were significantly smaller than the solid diet group which suggested growth retardation of the masticatory organ. 2. The submandibular gland wet weight and its DNA content in the liquid diet group were lower than in the solid diet group during the experimental period. This suggested that the number of cells and their ability to divide was diminished. The wet weight of glands per unit DNA content showed an increase in cell size during the adult period. 3. The alpha-amylase activity and total protein content of the submandibular glands in the liquid diet group were lower than in the solid diet group. This suggested that the saliva secretory function and protein synthesis were depressed, in agreement with the low number of cells detected. 4. The differentiation of the acinar cells and convoluted tubule cells of the liquid diet group was retarded and the 60 weeks subgroup showed hypertrophy of the convoluted tubule cells, disappearance of the secretory granules and a decrease in the number of acinar cells. 5. Slight stromal fibrosis was observed with increasing age (60 weeks) with no difference between the two groups. 6. This suggested that when mice were fed a liquid diet, the development of salivary glands was retarded and also aging changes occurred more rapidly.

Aging↗
PubMed2005

Submandibular gland transfer for prevention of xerostomia after radiation therapy: swallowing outcomes.

OBJECTIVE: To assess swallowing outcomes in patients with oropharyngeal carcinoma in relation to the Seikaly-Jha procedure for submandibular gland transfer (SJP). The SJP has recently been described as beneficial in the prevention of xerostomia induced by radiation therapy in patients with head and neck cancer. DESIGN: Inception cohort. SETTING: University-affiliated primary care center. PATIENTS: A phase 2 clinical trial was conducted from February 1, 1999, through February 28, 2002, to evaluate SJP in patients with head and neck cancer. During that period, a consecutive sample of 51 patients who underwent surgical resection and reconstruction with a radial forearm free flap for oropharyngeal carcinoma were referred for functional assessment of swallowing after completion of adjuvant radiation therapy. At 6 months after surgery, swallowing assessments for 24 patients were available. INTERVENTION: The cohort of 24 patients included 13 who had preservation of 1 submandibular gland (SJP group) and 11 who did not (control group). MAIN OUTCOME MEASURES: Quantitative and qualitative aspects of swallowing were obtained to determine whether patients in the SJP group performed more optimally than those in the control group. RESULTS: Baseline and stimulated salivary flow rates were significantly different between groups. Patients in the SJP group were able to move the bolus through the oral cavity and into the pharynx faster than those in the control group. In addition, patients in the SJP group swallowed less often per bolus than patients in the control group. The complete swallowing sequence was twice as long in controls. CONCLUSIONS: The SJP for submandibular gland transfer appears to be beneficial in promoting more time-efficient swallowing behaviors. This efficiency has implications for the overall well-being and nutritional status of patients with head and neck cancer.

Adult↗
PubMed1981

Effect of histamine on cyclic AMP levels in the submandibular gland.

Histamine and 4-methylhistamine increased on cyclic AMP level in chopped guinea-pig submandibular glands. 4-Methylhistamine was more active than histamine. 2-(2-Pyridyl)ethylamine was less active than histamine or almost inactive. Metiamide markedly antagonized the histamine-induced cyclic AMP increase but mepyramine had no inhibitory effect on this increase. These results suggest that there are H2-receptors which mediate the cyclic AMP response to histamine in the guinea-pig submandibular gland.

1-Methyl-3-isobutylxanthine↗
PubMed1989

[Effects of testosterone and estradiol-17 beta on the secretory granules of rat submandibular gland].

The effects of testosterone and estradiol-17 beta on the protein components, amino acid compositions and trypsin-like protease activities in the secretory granules of granular duct cells of rat submandibular glands were assayed. Male and female adult rats were divided into 4 groups, non treatment (control), castrated, testosterone injected and estradiol-17 beta injected groups. The castrated rats were received 5 injections of 5 mg/kg testosterone or 500 micrograms/kg estradiol-17 beta every other day 3 weeks after the castration. The secretory granules of submandibular gland obtained from each group were prepared using centrifugal fractionation with sucrose step gradient. The protein components of secretory granule were separated by 5-15% SDS-PAGE, and the SDS-PAGE revealed the characteristic proteins of MW 39,000 in male and MW 37,000 in female. The MW 39,000 and the MW 37,000 protein was disappeared by testectomy and ovaryectomy, respectively. The MW 39,000 protein was reappeared in the castrated male rat treated by testosterone and shown in castrated female rat treated by the same steroid. Similary the MW 37,000 protein was found in the castrated male and female rats treated by estradiol-17 beta. From the results of amino acid composition analysis, abundant Asp, Ser, Glu and Gly were revealed in the MW 39,000 protein and MW 37,000 protein. Trypsin-like protease activity located in secretory granule was reduced by castration in both male and female, however, the enzyme activities were elevated by testosterone or estradiol-17 beta injection, revealing rather higher activities than control. It was demonstrated that the secretory granule of rat submandibular gland contained the testosterone dependent protein having a MW of 39,000 and the estradiol-17 beta dependent protein having a MW of 37,000 and that the trypsin-like protease activities in the granules were sensitive to both testosterone and estradiol-17 beta.

Animals↗
PubMed2006

Sialoblastoma: a rare submandibular gland neoplasm.

A sialoblastoma is a rare congenital epithelial tumor that arises in a major salivary gland. To our knowledge, only 24 cases of sialoblastoma have been previously reported in the English-language literature. We report a new case, that of a 15-month-old boy who presented with a submandibular mass. Surgical excision of the mass was undertaken. Intraoperatively, the mass appeared to be adjacent to the submandibular gland, but it had not invaded it. The mass was excised, and the submandibular gland was left in place. Pathology identified the tumor as a sialoblastoma. However, pathology also revealed that residual tumor was present at the surgical margin. The patient was returned to the operating room for excision of the left submandibular gland and the level I lymph nodes. Following revision surgery, the surgical margins were negative. The patient remained disease-free at the 1-year follow-up. Despite the need for revision surgery, this case provides support for the idea that surgery alone is sufficient for curative treatment.

Follow-Up Studies↗
PubMed1981

Androgenic action of glucocorticoids on the granular ducts of mouse submandibular glands.

The administration of cortisol acetate and dexamethasone to castrated-adrenalectomized mice increased the submandibular gland weight, the granular duct cell size of the gland and the number of androgen-dependent granules detected with the staining for tryptophan in the duct cell. These changes were not as pronounced as when testosterone propionate was given. Similar effects on the duct cells and an increase in androgen-dependent esteroprotease activity of the submandibular glands were observed in castrated mice, but not in mice with testicular feminization which are genetically deficient in androgen receptors. These results suggest that glucocorticoids exert a certain degree of androgenic action on the granular duct cell through androgen receptors when androgens are absent or deficient.

Adrenalectomy↗
PubMed1986

Effects of substance P on glycoprotein secretion from acinar cells of the rat submandibular gland.

The action of substance P on glycoprotein secretion from acinar cells of the rat submandibular gland was described in this report. Salivation elicited by i.v. injection of 0.5 to 20 micrograms/kg of substance P was increased dose-dependently, and its flow rate was highest at the first 1 min. Major glycoprotein species secreted into saliva by substance P-stimulus were shown to be electrophoretically identical with those found in acini, but not granular convoluted tubules. These results support the view that substance P acts on acinar cells of the submandibular gland and stimulates secretion of saliva from the cells.

Animals↗
PubMed1982

Progressive secretory dysfunction in the rat submandibular gland after excretory duct ligation.

Unilateral ligation of the main excretory duct of the rat submandibular gland caused a progressive deterioration of secretory function characterized by: (1) the secretion of progressively smaller volumes of saliva in response to a standard maximal intraperitoneal (i.p.) dose of pilocarpine. Saliva volume was reduced 69.4, 88.8 and 95.9 per cent, respectively, 1, 3 and 7 days after duct ligation. By 2 weeks, the contralateral, non-ligated gland had an enhanced response to pilocarpine and the ligated gland a 96.8 per cent reduction in the volume of saliva secreted; (2) a progressive reduction in the maximum flow rates attained upon stimulation with pilocarpine, which were 23.4, 10.1 and 5.1 per cent of those attained in the contralateral gland at, respectively, 1, 3 and 7 days after ligation; (3) a progressive increase in the sodium concentrations of saliva, which became plasma-like 2 weeks after ligation; (4) a significantly reduced secretory response to standard doses of isoproterenol, phenylephrine, methoxamine and substance P 2 weeks after ligation. In addition, both cholinergic and adrenergic receptors were significantly reduced in number 2 weeks after ligation. Administration of daily injections of pilocarpine (10 mg, i.p.) after duct ligation reduced the gland atrophy observed 3 days (by 17 per cent) and 1 week (by 9 per cent) later, but did not prevent the reduction in volume or flow rats observed after duct ligation alone. Thus ligation of the main excretory duct causes a progressive dysfunction in acinar and duct cells in the rat submandibular gland and alters their responsiveness to physiological regulators of secretion. The alterations in glandular secretory function and in the composition of saliva which occur in diseases causing partial or complete obstruction to the flow of saliva are likely to be similar to the changes described.

Animals↗