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A morphologic study of the recovery of the rat submandibular gland after retrograde infusion. I. Water-soluble radiographic contrast medium.

The recovery of the rat submandibular gland after retrograde infusion of water-soluble radiographic contrast medium was studied using an experimental model. During continuous monitoring of the developing intraglandular pressure, the glands were subjected to ductal and slight parenchymal filling or heavy parenchymal filling with the medium. The animals were killed after varying recovery periods, and the tissue was prepared for light and electron microscopic examination. Dilation of the ductal lumina, induced during ductal and slight parenchymal filling, was successively reduced and, generally, the parenchyma had a normal appearance at 30 h. In glands subjected to heavy parenchymal filling, the changes in the intralobular ducts were more pronounced and were also seen at later times after infusion. Alterations in the acini, comprising fusion of secretory granules, vacuole formation and dilation of the acinar lumina and intercellular canaliculi, were observed. At later times, atrophy of the parenchymal cells occurred together with an apparent proliferation of the connective-tissue stroma, as well as an increase in the number of small blood vessels. An inflammatory cell-infiltrate was seen in both groups of animals, but was most prominent in glands subjected to heavy parenchymal filling. The infiltrate, comprised primarily of macrophages and polymorphonuclear leukocytes, reached a peak at 20 h after infusion. At later times, mast cells and occasional eosinophils were seen. The observed alterations and the pattern of recovery are most likely due to the induced intraglandular pressure and the following inflammatory reaction. It is also possible that the changes, to some extent, are influenced by the presence of the contrast medium in the tissue.

Animals↗

Vasoactive intestinal peptide interacts with alpha-adrenergic-, cholinergic-, and substance-P-mediated responses in rat parotid and submandibular glands.

Secretory dose-response curves were obtained with both acetylcholine and phenylephrine treatment in rat parotid and submandibular glands. Vasoactive intestinal peptide (VIP), which produced relatively low volumes of protein-rich saliva in rat salivary glands, also enhanced acetylcholine-, phenylephrine-, and substance-P-mediated fluid and protein secretion when administered in combination with these agents. The specific mechanisms involved in the synergistic actions of VIP with substances such as acetylcholine, phenylephrine, and substance P, which are primarily linked to the production of fluid secretion in rat salivary glands, have yet to be determined.

Acetylcholine↗

Sialogogic effects on rat submandibular gland of analogs of the C-terminal hexapeptide of substance P.

The sialogogic response of submandibular glands to analogs of the C-terminal hexapeptide of substance P with various amino acids at the N-terminus was investigated in urethane-anesthetized rats. The rank order of potencies was as follows: SP greater than (pGlu6)SP6-11 much greater than (Dab6)SP6-11 greater than (Orn6)SP6-11 greater than (Gln6)SP6-11 greater than (Lys6)SP6-11 much greater than (Ala6)SP6-11. These results suggest that the sialogogic activity of the analogs of the C-terminal hexapeptide is influenced by the steric effects of the N-terminal amino acid, and the nature of its side chain is of particular importance.

Animals↗

Rapid purification of tonin, esterase B, antigen psi and kallikrein from rat submandibular gland by fast protein liquid chromatography.

Tonin, esterase B, antigen psi and kallikrein from the rat submandibular gland were purified by fast protein liquid chromatography with Mono P or Mono Q columns. The purity of the separated proteins was evaluated by sodium dodecyl sulphate polyacrylamide gel electrophoresis and by isoelectrofocusing in flat-bed polyacrylamide gel. Tonin and esterase B were purified by DE-52 cellulose anion-exchange chromatography and chromatofocusing on Mono P in two and three steps, respectively. Antigen psi and kallikrein were purified by a two-step procedure using DE-52 cellulose and Mono Q anion-exchange chromatography. The high resolution power of Mono Q revealed the different isoenzymes of kallikrein.

Animals↗

Ultracytochemical detection of heteropolysaccharides in the cat submandibular gland.

We have investigated the morphology and the ultrastructural distribution of complex carbohydrates in the cat submandibular gland using the high iron diamine-thiocarbohydrazide-silver proteinate (HID-TCH-SP), the low iron diamine-thiocarbohydrazide-silver proteinate (LID-TCH-SP), and the periodate-thiocarbohydrazide-silver proteinate (PA-TCH-SP) sequences. The results of morphological analyses show that, in addition to already known cellular populations, in striated ducts a further cellular type containing glycogen is present. The histochemical staining showed that demilunar cells secrete polysaccharides that have carboxylated but not sulphated glycosidic residues, whereas acinar cells elaborate secretion products rich in carboxylated and sulphate-containing complex carbohydrates. Vicinal glycol groups are present as constituents of the secretion in demilunar, acinar, and ductal cells.

Animals↗

New observations on the innervation of striated ducts in submandibular glands of cats, including possible peptidergic nerves.

The presence of hypolemmal axons between striated duct cells in submandibular glands of cats has been established electron microscopically. Axons were found between "light" cells, between "light" and "dark" cells and between "light" and basal cells. Hypolemmal axons were observed most frequently in the junctional region between striated and intercalary ducts. They were often more common in younger animals. "Dark" cells with numerous processes sometimes appeared to have a special relationship with hypolemmal axons. Most of the hypolemmal axons in striated ducts contained characteristic agranular vesicles of the cholinergic type, about 40 nm in diameter; many of these axons also contained large dense cored vesicles of the peptidergic type, about 100 nm in diameter and possessing a more clear outer halo. No adrenergic axons have been observed beneath the basal lamina of striated ducts, even after use of 5-OHDA. The possibility that some of the hypolemmal axons in striated ducts are peptidergic and their possible functions are discussed. Apart from other activities these axons may have a role in supplying special trophic factors to the cells, helping them in their developmental specialisation and maintaining them in normal condition. An absence of such factors after parasympathetic decentralisation may be responsible for the dramatic atrophic changes in striated duct cells, especially since the atrophy in the gland is not solely due to an absence of acetylcholine activation.

Animals↗

Conversion from 4S androgen receptor from rat submandibular gland to higher molecular form and the effect of sodium molybdate.

The androgen receptor from rat submandibular gland was transformed by exposure to ATP at 0 degrees C. The transformed 4 S receptor converted to a higher molecular weight form in a low-salt glycerol gradient centrifugation when ATP was removed from the sample. The sedimentation coefficient of the converted receptor was similar in the absence or presence of 20 mM molybdate; 7.8 +/- 0.5 S without molybdate and 7.6 +/- 0.3 S with molybdate. However, the receptor converting in the presence of molybdate could markedly bind to DNA-cellulose, while an entity without molybdate could not. These results suggest that molybdate directly interacts with the DNA-binding domain on the 4 S androgen receptor and prevents this domain from being concealed by conversion in low-salt conditions.

Adenosine Triphosphate↗

Salivary secretion of immunoglobulin A by submandibular glands in response to autonomimetic infusions in anaesthetised rats.

Salivary secretion of immunoglobulin A (lgA) by submandibular glands is increased by stimuli from autonomic nerves. Since it is unclear which specific autonomic receptors transduce such stimuli, we have infused autonomimetics intravenously and compared secretion of fluid, IgA and stored proteins (peroxidase and total protein) with secretory responses during electrical stimulation of the parasympathetic nerve supply in anaesthetized rats. The greatest secretion of IgA was evoked by the alpha-adrenoceptor agonist phenylephrine and this was reduced by the beta-adrenoceptor blocking drug propranolol. The secretion of fluid or proteins but not IgA was increased with frequency of nerve stimulation and dose of methacholine (cholinergic), isoprenaline (beta-adrenergic) or phenylephrine (alpha-adrenergic).

Adrenergic Agonists↗

Glycoprotein secretion in the mouse submandibular gland as revealed by radioautography after L-3H-fucose injection.

Glycoprotein secretion in the mouse submandibular gland was investigated by light microscope radioautography of semi-thin sections after the administration of L-3H-fucose. The incorporation of the precursor in the acini was negligible. 3H-fucose was taken up in the paranuclear region of the cells lining the intercalated, secretory, striated and excretory ducts. This labeling pattern was interpredted as addition of the precursor to glycoproteins within the Golgi apparatus. Incoropration in the intercalated duct was restricted to the cells with fine cytoplasmic granules. The glycoproteins synthesized by the intercalated and secretory ducts were transported to the saliva by the secretion granules. It is assumed that the glycoproteins synthesized in the striated and excretory ducts are plasma membrane glycoproteins which seem to renew continuously. Quantitation of the radioautographs supplied data concerning the incorporation of 3H-fucose into newly synthesized glycoproteins as well as the renewal of the labeled macromolecules in each duct.

Animals↗

Autonomic regulation of tight junctional permeability in the rat submandibular gland.

The permeability of tight junctions to tracers having different molecular weights was investigated in the submandibular gland of rats stimulated parasympathetically, sympathetically, or both. Lactoperoxidase (82,000 daltons), horseradish peroxidase (40,000 daltons), and microperoxidase (1,630 daltons) were used as the tracers. The tracers were administered by close arterial infusion via the glandular artery, and their secretion into the saliva was quantified biochemically, and their secretory routes within the gland were determined histochemically at the electron microscopic level. Microperoxidase and horseradish peroxidase passed into the saliva by electrical stimulation of either the chorda or superior cervical ganglion, and the combined stimulation of both caused a larger output of both tracers. No output of lactoperoxidase into the saliva occurred with any type of nerve stimulation. Electron microscopic histochemical observations showed that molecules of molecular weight equal to or lower than that of horseradish peroxidase entered the lumen through the tight junctions between adjacent acinar cells following combined stimulations of chorda and superior cervical ganglion. These findings indicate that electrical stimulation of parasympathetic and sympathetic nerves causes an increase in tight junctional permeability of acinar cells to microperoxidase and horseradish peroxidase. Although the combined stimulation of parasympathetic and sympathetic nerves resulted in increased junctional permeability to these tracers, the junctions remained impermeable to larger molecules, i.e., lactoperoxidase.

Animals↗

Alteration by ouabain of rat submandibular glands function.

1. Effects of various doses of intraperitoneal ouabain (1,2 and 5 mg/kg) on rat submandibular saliva were investigated in this study. 2. Potassium and calcium and their product (K+ x Ca2+) were found to be elevated in all groups. 3. Changes in salivary flow were not the major cause of the alterations in electrolytes. 4. Protein concentrations were elevated in the doses of 1 and 2 mg/kg of the drug and somewhat reduced in the dose of 5 mg/kg of ouabain but still above the base line. 5. The results show that there is an ouabain-induced close parallelism between magnesium and total protein secretion from rat submandibular glands.

Animals↗

Lethal factor in the male mouse submandibular gland.

The presence of a toxic component (lethal factor) has been reported in the male mouse submandibular gland (SMG). This study was designed to clarify whether the toxic component was secreted into saliva or not and to investigate some characteristics of the component. Not only SMG extract but also phenylephrine-induced saliva from male mice was highly toxic to guinea pigs, rats, hamsters, and mice. The toxic activity of the saliva was found in all strains of male mice examined. In female mice, the toxic activity was extremely low. One of the toxic components in the saliva was purified by isoelectric focusing and DEAE Sephadex A-50 chromatography and identified with a kallikrein-like enzyme. The activity of the enzyme markedly increased in saliva secreted to alpha-adrenergic agents but not to beta-adrenergic or cholinergic agents. These results indicate that the lethal factor in the male mouse SMG is an exocrine protein with Kallikrein activity and its secretion into saliva is controlled through alpha-adrenergic receptors.

Animals↗

Parasympathetic reflex vasodilatation in rat submandibular gland.

The present study was designed to investigate 1) whether parasympathetic reflex vasodilatation occurs in the submandibular gland (SMG) in deeply urethan-anesthetized, cervically vagotomized, and sympathectomized rats when the central cut end of the lingual nerve (LN) is electrically stimulated and 2) to what extent the neural mechanisms underlying such responses are the same as those involved in the response to direct stimulation of the chorda-LN (CLN). Stimulation of each nerve separately elicited a marked blood flow increase in SMG. Section of the chorda tympani abolished the SMG blood flow response but had no effect on the lip blood flow increase evoked by LN stimulation. Section of the CLN abolished the SMG blood flow increases evoked by stimulation of either nerve. The SMG blood flow increases (regardless of whether they were evoked by LN or CLN stimulation) were markedly reduced by the autonomic cholinergic ganglion blocker hexamethonium. The present study demonstrates that a parasympathetic reflex vasodilator mechanism is present in the rat SMG and that it can express its effects under deep general anesthesia.

Afferent Pathways↗

Reversal of relative proteinase F activity and onset of androgen-dependent proteinases in the submandibular gland of postnatal mice.

By isoelectric focusing, we separated trypsin-like proteinases of the mouse submandibular gland (ICR strain) into isozymes with pI values of 4.6 (proteinase F), 5.6 (protease D), 5.8 (protease A), 7.1 and 9.9 (P-esterase). During postnatal development, proteinase F appeared earliest (on the 15th day after birth) and increased in both sexes; however, its percentage ratio to total activity decreased markedly with time because of the rapid increase of other proteinases. On the 22nd day of life, proteinases A and D appeared, and the increase of a proteinase with pI-7.1 followed thereafter. P-esterase was the last isozyme to appear, becoming detectable around 29-45 days. After maturation, the activities of protease A plus D, P-esterase, and the isozyme with a pI value of 7.1 were higher in males than in females, whereas the relative level of proteinase F was reversed. We conclude that proteinase F is appreciably different from the other four proteinases in its development pattern as well as in its responsiveness to sexual hormones.

Androgens↗

Giant salivary calculi: an orocervical fistula caused by a submandibular gland calculus.

Large calculi of the salivary glands are rare. They may go undetected for many years. The authors report three cases of giant submandibular gland calculi. In one patient, the calculus was an incidental finding. In all patients, the mass was in the substance of the gland; in one, the configuration of the mass was unusual and it had eroded through the floor of the buccal cavity forming an orocervical fistula (the first such report) and in another the mass occupied a small portion of Wharton's duct. The histopathologic findings in all three cases were of nonspecific chronic inflammation. All the patients made a smooth recovery and had no complaints at follow-up.

Adult↗

Effects of reserpine treatment on beta-adrenergic/adenylate cyclase modulate secretion and resynthesis by the rat submandibular gland.

Chronic reserpine (adrenergic blocking) treatment causes a marked accumulation of secretory protein in the rat submandibular gland (SMG) but discharge of this material is delayed in response to isoproterenol stimulation. The purposes of this study were to investigate the effects of chronic reserpine treatment on 1) the number of beta-adrenergic receptor. 2) the sensitivity of cell-surface-associated adenylate cyclase to various concentrations of isoproterenol, and 3) to correlate these data to morphologic studies of the secretion and resynthesis phases of the isoproterenol-induced secretory cycle in the rat SMG. Animals were injected with reserpine (0.5 microgram/g b.w.) for 6 days. Plasma membrane fractions were prepared. The adenylate cyclase response to a series of isoproterenol concentrations, and the number of beta-adrenergic receptors ([3H]-alprenolol binding) were determined. Other animals were given a single dose of isoproterenol (0.8 mg/100 g b.w.) and the SMG was examined by light and electron microscopy at various times (30 min to 24 h) after treatment. Chronic reserpine treatment leads to a 2.5-fold increase in SMG beta-adrenergic binding sites and a 50-fold increase in adenylate cyclase sensitivity to IPR stimulation when compared to controls. However, secretion and resynthesis of secretory product in response to IPR stimulation was greatly delayed in reserpinized rats.

Adenylyl Cyclases↗

[TNM classification system in cancer of the submandibular gland--a comprehensive retrospective study of 271 primary cases].

UNLABELLED: A total of 271 patients with submandibular gland cancer, treated in 149 hospitals in Japan in the period from 1958 to 1991, were retrospectively studied with regard to age, sex, TNM classification (UICC 1987, Geneva), histological diagnosis, therapeutic method and prognosis. The results were the following: 1. 157 males and 114 females 2. Stage I: 52 cases (19%); stage II: 64 cases (24%); stage III: 52 cases (19%) and stage IV: 103 cases (38%). 3. T1: 29 cases (11%); T2:116 cases (43%); T3: 93 cases (30%) and T4: 43 cases (16%). 4. Cervical lymph node involvement: 40%, ranging 25% in adenoid cystic carcinoma and acinic cell tumor, to 62% in undifferentiated carcinoma. 5. Distant metastasis: 11% 6. HISTOLOGY: adenoid cystic carcinoma 37%, adenocarcinoma 20%, mucoepidermoid tumor 16%, carcinoma in pleomorphic adenoma 10%, epidermoid carcinoma 10%, undifferentiated carcinoma 3%, acinic cell tumor 3%. 7. Treatment method: Surgical procedure alone; 51%, Combination of surgery and postoperative radiation; 18%. 8. 5-year and 10-year survival rates were 36% and 11%, respectively. 9. 5-year and 10-year survival rates varied according to the stages, being 76% and 38%, respectively, for stage I, 68% and 20%, respectively, for stage II, 15% and 10%, respectively, for stage III, and 14% and 4%, respectively for stage IV.

Adolescent↗

Activation by P2X7 agonists of two phospholipases A2 (PLA2) in ductal cells of rat submandibular gland. Coupling of the calcium-independent PLA2 with kallikrein secretion.

Isolated ductal cells of rat submandibular gland phospholipid pools were labeled with [3H]arachidonic acid (AA). The tracer was incorporated preferentially to phosphatidylcholine (46% of the lipidic fraction). Extracellular ATP induced the release of [3H]AA to the extracellular medium in a time- and dose-dependent manner (EC50 = 220 microM). Among other agents tested, only 2', 3'-O-(4-benzoylbenzoyl)adenosine 5'-triphosphate (Bz-ATP) was able to mimic the effect of ATP (EC50 = 15 microM), without activation of phospholipase C. The purinergic antagonists oxidized ATP, suramin, and Coomassie Blue partly inhibited the response to 1 mM ATP and 100 microM Bz-ATP; the response was also blocked by the addition of Mg2+ or Ni2+. Expression of P2X7 receptor mRNA in these cells was confirmed by reverse transcription-polymerase chain reaction. In the presence of extracellular calcium, the phospholipase A2 inhibitor 2-(p-amylcinnamoyl)amino-4-chlorobenzoic acid (a nonspecific inhibitor), arachidonyl trifluoromethylketone (AACOCF3, an inhibitor of the calcium-dependent cytosolic PLA2 (cPLA2)), and bromoenol lactone (an inhibitor of the calcium-independent PLA2 (iPLA2)) inhibited the release of [3H]AA induced by ATP and Bz-ATP. In the absence of extracellular calcium, the release of [3H]AA in response to the purinergic agonists was still observed; this response was not affected by AACOCF3 and completely blocked by bromoenol lactone. ATP and Bz-ATP stimulated a calcium-independent secretion of kallikrein, which could be blocked by BEL but which was enhanced by AACOCF3. It is concluded that the P2X7 receptor in ductal cells is coupled to kallikrein secretion through a calcium-dependent cPLA2 and a calcium-independent iPLA2.

Adenosine Triphosphate↗