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PubMed1985

Acquisition of urine, kidney and submandibular gland epidermal growth factor responsiveness to thyroxine administration in neonatal mice.

Thyroxine (T4) administration to newborn mice on days 0-6 produced no measurable changes of submandibular gland (SMG) or kidney epidermal growth factor (EGF) concentrations on day 7, compared with vehicle-treated control pups. By contrast, this regimen caused a large increase in urine EGF levels. The effects of three T4 injection regimens (on days 0-6, 7-14 and 0-14) were then studied on day 15. The 0-14 and 7-14 day-regimens elicited large increments in urine, SMG and kidney EGF concentration, the 0-14 day-regimen having the greatest effect. The 0-6 day-regimen had no effect on SMG and kidney EGF concentrations on day 15 but did increase urine EGF. In summary, urine EGF responsiveness to T4 is present during the first week of life, but SMG and kidney EGF responsiveness develops only in the second week. Administration of T4 appears to accelerate the normal ontogeny of urine, kidney and SMG EGF.

Aging↗
PubMed1979

Isolation of an acidic protein from the armadillo submandibular gland.

An acidic protein fraction with an apparent molecular weight of 34 000 has been isolated from the Cetavlon-treated, mucin-free supernatant of the armadillo submandibular gland 0.01 M NaCl extract. This purified material, which was obtained in a yield of 0.45%/g wet gland, contains 24 mol % acidic amino acids and 4 mol % basic amino acids. Hexosamines, sialic acid, and neutral sugars represent 7% of the dry sample weight. In polyacrylamide gel and cellulose acetate electrophoresis, a single protein band was observed. The acidic protein fraction is highly reactive with the Lowry phenol reagent, giving a protein value 83% higher than that obtained by summation of its anhydrous amino acids, and is explained by the occurrence of peptide linkages peculiar to this material. The presence of other basophilic components besides mucus glycoproteins within the salivary gland of the armadillo may have physiological significance.

Amino Acids↗
PubMed1975

Two phases of chorda-lingual induced vasodilatation in the cat's submandibular gland during prolonged perfusion with Locke solution.

The effect of stimulation of the chorda-lingual nerve on the venous flow has been studied in cat submandibular glands perfused with Locke solution for 2-4 hr. When trains of pulses at 25 Hz were given for 1-5 sec, two distinct phases of vasodilatation were observed: a rapid initial phase of high amplitude and a slower developing more prolonged phase of smaller amplitude. Repeated stimulations did not lead to a reduction of the vasodilatory response. A close relationship was found between the duration and magnitude of the second phase of vasodilatation and the duration and magnitude of the post-stimulatory, active reuptake of potassium. When the active reuptake of potassium was prevented either by ouabain (which inhibits active transport) or by atropine (which abolishes the stimulation induced loss of potassium) the second phase of vasodilatation was severely reduced, while the initial phase remained virtually normal. It is concluded that the initial phase of vasodilatation probably is mediated by vasodilator nerve fibres. The second phase is perhaps causally related to the post-stimulatory, active transport of cations. An involvement of bradykinin formation is highly unlikely under the given experimental conditions.

Animals↗
PubMed1999

Regulated permeability of tight junctions by E-cadherin in rat submandibular gland acini.

We examined the relationship between E-cadherin (E-CD), protein kinase C (PKC) and assembly of the tight junction (Tj) in rat submandibular gland acinar cells. The junctional complex of the acinar cells was double-labeled with anti-ZO-1 antibody and anti-E-CD antibody. When Ringer's solution was intraductally injected into the main duct, ZO-1 labels were highly concentrated at the Tj zone, and the adherens junction (Aj) was exclusively labeled by E-CD. In addition, the Tj was impermeable to microperoxidase. Neither intraductal injection of anti-E-CD antibody solution nor infusion of carbachol produced labels for ZO-1 at the Tj zone, although these proteins were occasionally intermixed at the Aj zone. In these cases, the Tj was permeable to microperoxidase. Intraductal injection of anti-E-CD antibody solution with PKC agonist, resulted in a reduction of E-CD labels in the Aj zone, while the ZO-1 was labeled exclusively in the Tj zone. In this case, Tj was impermeable to microperoxidase. These results suggest that E-CD plays a major role in mediating intercellular physical adhesion, and that PKC may be active in signaling the pathway activated by E-CD-mediated cell-cell adhesion.

Animals↗
PubMed1992

Effect of substance P administration on vascular permeability in the rat dental pulp and submandibular gland.

The effects of substance P (SP) administration on vascular permeability were studied in the dental pulp (DP) of upper and lower incisors and in the submandibular gland (SMG) of male rats. Vascular permeability was assessed by means of extravasation of Evans blue dye. SP was diluted in 0.5% bovine serum albumin (BSE) and infused into the left common carotid artery. Separate groups of animals receive chloropyramine, an H1 histamine receptor antagonist (10 mg kg-1 i.v.) or indomethacin, a prostaglandin synthesis inhibitor (4 mg kg-1 i.v.) prior to SP infusions. Infusion of SP for 5 min increased plasma extravasation both in DP and SMG, with a threshold of about 30 pmol min-1 and 74 pmol min-1, respectively. Enhanced salivary secretion was also observed. Although the administration of 74 pmol min-1 of SP significantly lowered the systemic blood pressure, experimental hypotension elicited by haemorrhage did not influence vascular permeability in either organ tested. After chloropyramine administration the SP effect on vascular permeability in both DP and SMG was abolished. Indomethacin pretreatment failed to prevent the permeability-enhancing action of SP. Our results suggest that substance P increases both pulpal and glandular plasma extravasation in the rat indirectly, via the release of histamine and the activation of H1 histamine receptors.

Animals↗
PubMed1986

Effect of adrenoceptor blockers on the acinar enlargement induced by Chagas' disease in the rat submandibular glands.

Chagas' disease induces acinar enlargement in the submandibular salivary gland of rats inoculated with the Y strain of Trypanosoma cruzi at the age of 27-28 days and killed 18 days after inoculation (acute phase). Propranolol, a beta-adrenoceptor blocker, prevents the acinar enlargement induced by T. cruzi infection. Phenoxybenzamine, an alpha-adrenoceptor blocker, enhances this enlargement. These results suggest that catecholamines are involved in the genesis of the acinar hypertrophy induced by Chagas' disease.

Animals↗
PubMed2003

Growth of cell populations of the intralobular duct in the submandibular gland of the mouse during postnatal development.

The growth of cell populations of intercalated ducts (ID), striated ducts (SD) and convoluted granular tubules (CGT) of the submandibular gland of the mouse was analyzed, during the period of 14 to 84 days of postnatal life, by means of morphometric methods. The absolute number of cells in each type of duct was submitted to curve fitting by means of linear regression analysis, and the obtained equations were: y = 13.22 + 0.87x for ID cells in the growth period of 14 to 84 days; y = -18.82 + 3.69x and y = 176.03 - 1.93x for SD cells in the periods of growth and decay of cells, from 14 to 35 and 35 to 84 days, respectively; and y = 90.59 + 4.60x for CGT cells in the growth period of 28 to 84 days. Based on these equations, the daily accumulation (growth rate) or loss (decaying rate) of cells in the period covered by each equation was calculated. Thus, the rate of growth of ID cells was 0.87 x 10(5) cells/day; the growth and decline rates of SD cells were 3.69 x 10(5) cells/day and -1.93 x 10(5) cells/day, respectively; and the growth rate of CGT cells was 4.60 x 10(5) cells/day. The analysis of the accumulation or loss of cells and the balance of cell populations in the various types of ducts revealed that SD cells were transformed into CGT cells and that the growth of these cells population also depends on the formation of new cells through proliferative activity, probably in the intercalated ducts.

Animals↗
PubMed1989

[Study on the translocation of cytosolic dihydrotestosterone-androgen binding protein complex to the nuclei in rat submandibular gland].

The low molecular weight inhibitor (LMWI) and the translocation of dihydrotestosterone (DHT)-androgen binding protein (ABP) complex of the cytosol to the nuclei in rat submandibular gland (SMG) was studied by dialysis, ultrafiltration and glycerol linear gradient centrifugation procedures. Prebound cytosol was obtained by the incubation with 3H-DHT at 4 degrees C for 3hr in the presence or absence of 100 fold excess of unlabeled DHT prior to contact to nuclei. When prebound cytosol was dialyzed or ultrafiltrated, the binding ability of 3H-DHT-ABP complex to nuclei was increased up to 3 times of the control (nontreated prebound cytosol). The sedimentation rate of 3H-DHT-ABP complex by glycerol linear gradient centrifugation was 4S for dialyzed or ultrafiltrated prebound cytosol and 8S for control. These transformation of 8S to 4S and activation to the nuclear binding of 3H-DHT-ABP complex by dialysis or ultrafiltration were inhibited by molybdate in the prebound medium. The similar transformation of 8S to 4S and activation to the nuclear binding of 3H-DHT-ABP complex was shown in heated prebound cytosol. These results indicate that the LMWI regulate activation of 3H-DHT-ABP complex. The molecular weight of the dialyzed LMWI were estimated as 7,000 by dialysis or ultrafiltration membrane and SDS-PAGE. From the in vitro two step binding assay, it was revealed that rat SMG contained the low molecular weight protein (M. W. 7,000) in cytosol having a inhibitory effect on the translocation to the nuclei of 3H-DHT-ABP complex.

Androgen Receptor Antagonists↗
PubMed2004

Increases in nerve growth factor immunoreactivity in the submandibular gland, but not in the parotid gland, of the rat following sympathetic denervation.

The effect of sympathetic denervation on nerve growth factor (NGF) immunoreactivity in the submandibular and parotid glands of adult female rats using a two-site, enzyme-linked immunosorbent assay was investigated. Unilateral sympathetic denervation through the avulsion of the superior cervical ganglion resulted in elevated levels of total amounts (decreased from 40-20%) and concentrations (decreased from 45-32%) of NGF in the submandibular glands, but not in the parotid glands, on the operated side 7-28 days post-operatively. Sympathetic decentralisation had no effect. The present findings support the hypothesis that increased levels of NGF play an important part in the increased activity of the acetylcholine-synthesising enzyme, choline acetyltransferase, in the rat submandibular gland following sympathetic denervation.

Animals↗
PubMed1996

Comparative study of kallikrein-like serine proteinases from rat submandibular glands.

The present investigation describes the comparative properties, particularly the substrate specificity of three kallikrein-like serine proteinases (I, II and III) purified from rat submandibular gland extract (Bedi, G.S., Prep. Biochem. 22,67-81, 1992). The physico-chemical and immunological properties of three proteinases were compared by Western blot analysis, immunodiffusion, immuno-electrophoresis, amino terminal sequence analysis, molecular weight determination and isoelectric focusing. Detailed substrate specificity of these proteinases was determined using chromogenic substrates, synthetic peptides and native proteins. The chromogenic substrate tosyl-gly-pro-arg-pNA was hydrolyzed preferentially by Proteinase I. The replacement of pro at the P2 position with bulky hydrophobic residues phe and leu completely abolished the hydrolysis by Proteinase I. The hydrolysis of the chromogenic substrates by Proteinase II was also affected by the amino acid residue present at the P2 position in the order of pro > gly > val > leu > phe. Neither Proteinase I nor Proteinase II hydrolyzed substrates in which arg was replaced with lys at the P1 position. Proteinase III was reactive against all the chromogenic substrates with arg or lys at the P1 position. Synthetic polypeptides T-kinin-leu and insulin B chain were resistant to cleavage by both Proteinase I and II and were cleaved specifically at arg-X peptide bond by Proteinase III. Tonin-like activity of Proteinase II was confirmed by cleavage of the angiotensin 1-14 at phe-his linkage to generate two fragments DRVYIHPF and HLLVYS respectively. All three proteinases cleaved human high molecular weight kininogen but only Proteinase III could cleave T-kininogen. Proteinase III was also reactive towards human and bovine fibronectin, fibrinogen and gelatin. Several other salivary and serum proteins were resistant to cleavage by these proteinases. Although the three enzymes are immunologically related, they differ with respect to size, isoelectric point, amino terminal sequence and inhibition profile.

Amino Acid Sequence↗
PubMed2006

Tissue kallikrein mK13 is a candidate processing enzyme for the precursor of interleukin-1beta in the submandibular gland of mice.

By using Western blot analysis, high levels of 17.5- and 20-kDa interleukin-1beta (IL-1beta) proteins were detected in the submandibular gland (SMG) of mice. Despite this fact, the amount of pro-IL-1beta protein, a precursor of IL-1beta, with a molecular size of 35 kDa in this tissue was below the detectable level, although strong expression of pro-IL-1beta mRNA was observed. A large amount of 17.5-kDa IL-1beta also appeared in the saliva of mice injected with lipopolysaccharide, suggesting that this IL-1beta is a secretory form produced by the SMG. The protein for IL-1beta-converting enzyme, a processing enzyme for pro-IL-1beta, was expressed only at a low level in the SMG as compared with its level in various epithelial tissues or lipopolysaccharide-stimulated macrophages. On the other hand, mK1, mK9, mK13, and mK22, members of the kallikrein family, were detected strongly in the SMG but not in other tissues. By incubation with mK13, but not with mK1, mK9, or mK22, the 35-kDa pro-IL-1beta was cleaved into two major products with molecular masses of 17.5 and 22 kDa, and production was inhibited by phenylmethylsulfonyl fluoride, a serine protease inhibitor, but not by IL-1beta-converting enzyme inhibitors. A peptide segment corresponding to amino acid residues 107-121 of mouse pro-IL-1beta (107WDDDDNLLVCDVPIR) was cleaved by incubation with mK13, generating two peptides, 107WDDDDNL and 114LVCDVPIR. Therefore, kallikrein mK13 would appear to hydrolyze pro-IL-1beta between its Leu113 and Leu114 residues. The results of immunohistochemistry and an autonomic therapy experiment showed that IL-1beta and kallikrein mK13 were co-localized in the secretory granules of granular convoluted tubular cells. Our present results thus suggest kallikrein mK13 is a plausible candidate for the processing enzyme for pro-IL-1beta in the SMG of mice.

Animals↗
PubMed1999

The protein phosphatase inhibitors, okadaic acid and calyculin A, induce apoptosis in human submandibular gland ductal cell line HSG cells.

OBJECTIVE: To investigate a possible relationship between protein phosphorylation or dephosphorylation status and apoptosis in salivary gland cells, we examined the effects of okadaic acid and calyculin A, the protein phosphatase inhibitors, on cultured human submandibular gland ductal cell line, HSG cells. METHODS: HSG cells at subconfluent stages were exposed to varying concentrations of okadaic acid or calyculin A. Apoptoses were analysed in HSG cells by phase-contrast microscopy, WST-1 cytotoxicity assay, Hoechst 33342 staining, and DNA ladder formation. RESULT: Both okadaic acid and calyculin A induced cell death in HSG cells in a dose-dependent fashion. Marked nuclear condensation and fragmentation of chromatin was observed in HSG cells. DNA ladder formation was also detected in HSG cells by treatment with okadaic acid or calyculin A. The induced DNA ladder formation was dose-dependent with maximal effect at concentrations of 50 nM okadaic acid and 2 nM calyculin A, respectively, and were time-dependent from 14 h to 48 h. To further determine if new gene transcription and protein synthesis regulate okadaic acid-induced apoptosis in HSG cells, the cells were treated with cycloheximide or actinomycin D in the presence of 20 nM okadaic acid. Neither inhibitor protected the cells against okadaic acid-induced apoptosis. CONCLUSION: Based on the known selectivity of okadaic acid and calyculin A, our results indicate that the pathway of the apoptosis in the cultured salivary gland cells is regulated by protein phosphatase type 1 or type 2A. Our results also suggest that new protein synthesis and/or mRNA expression are not involved in okadaic acid-induced apoptosis in HSG cells.

Apoptosis↗
PubMed1990

Effect of testosterone on proteoglycans in the submandibular glands of the rat.

The proteoglycans in the submandibular salivary glands of castrated male Wistar rats were studied before and after the daily administration of testosterone propionate (TP) for one month. Castration decreased the weight of the glands and their uronic acid content. The administration of TP reversed these effects. Chromatographic separation of the uronic acid fractions was performed on cellulose microcolumns. The principal fractions were hyaluronic acid, heparan sulfate and dermatan sulfate. There were also changes in the physical properties of the proteoglycans. Castration decreased the range of distribution of molecular weight and the density, while the lateral chains of smaller length disappeared. TP administration to castrated rats reversed these effects.

Animals↗
PubMed1982

Further evidence for postganglionic actions of prostaglandin F2 alpha and histamine in the submandibular gland of the dog.

To test the possibility that prostaglandin F2 alpha and histamine might produce salivary and vasodilator responses by stimulation of preganglionic parasympathetic nerve fibers in the submandibular gland the responses to the two substances of chronically decentralized glands were compared with those of acutely decentralized ones in anesthetized dogs. No significant differences in responses to either prostaglandin F2 alpha or histamine were noted between chronically decentralized glands and acutely decentralized ones. Thus, it was concluded that prostaglandin F2 alpha and histamine only stimulate parasympathetic postganglionic neurons within the gland and neither of the responses is related to non-cholinergic ganglionic transmission.

Acetylcholine↗
PubMed2007

Temporospatially regulated expression of subtilisin-like proprotein convertase PACE4 (SPC4) during development of the rat submandibular gland.

The temporospatial expression of PACE4, a member of the mammalian subtilisin-like proprotein convertase family involved in the activation of growth/differentiation factors, was investigated by in situ hybridization during the development of the rat submandibular gland (SMG). At the initiation stage (day 15.5 of gestation; E15), PACE4 was intensely expressed in the submandibular epithelium, but weakly expressed in the mesenchymal cells. At E16 when the branching morphogenesis becomes obvious, the expression of PACE4 in the mesenchyme was further decreased, although its level in the submandibular epithelium had not changed remarkably from that at E15. During the next stage of embryonic development (E17-E20), PACE4 was expressed in the cells derived from the submandibular epithelium, which include the proacinar, terminal tubular, and presumptive ductal cells. In the perinatal SMG, PACE4 was still expressed intensely in the terminal portion of the SMG containing the proacinar and terminal tubular cells, whereas its expression in the ductal cells was obviously decreased at the second postnatal day (P2) and at P6. Acinar cells expressing no PACE4 appeared, and their numbers increased following their development (P9-P20). At P30 when the PACE4 expression in the acinar cells was completely suppressed, its expression in the ductal cells became intense again. This temporospatially regulated expression of PACE4 suggests its apparent association with the proliferation, differentiation, and establishment of functional acinar and ductal cells of the SMG.

Animals↗
PubMed1997

Submandibular gland peptide-T (SGP-T) inhibits intestinal anaphylaxis.

A novel peptide, submandibular gland peptide-T (SGP-T), which reduces allergen-induced hypotension, was examined for effects on intestinal anaphylaxis. Hooded-Lister rats were sensitized to egg albumin and prepared for the measurement of in vivo myoelectric activity of the jejunum. The disruption of migrating myoelectric complexes (MMCs) that occurs upon intraluminal, duodenal challenge with antigen of sensitized rats was inhibited by 75% upon intravenous treatment with 100 micrograms/kg of SGP-T. In addition, SGP-T reduced the number of rats experiencing anaphylactic diarrhea and disrupted MMCs, but the peptide did not alter antigen-provoked release of rat mast cell protease II. The mechanism of action of SGP-T remains to be determined, but it apparently does not act directly on mast cells to exert its antianaphylactic action. These results emphasize that modulation of immediate hypersensitivity reactions is only one of several gastrointestinal activities that are affected by growth factors and peptides released from salivary glands.

Anaphylaxis↗
PubMed1978

Postnatal development and differentiation of the opossum submandibular gland.

The postnatal development and differentiation of the submandibular salivary gland has been examined in sixteen groups of young opossums. At birth the glandular elements, dispersed in loose connective tissue, consist only of ducts that are immature in appearance and of irregular secretory end-pieces. Development occurs in two phases, the first from birth to approximately 31 days postnatum, and the second thereafter. During the first phase the ductular elements show separation into intercalated and intralobular ducts, and attain structural maturity. The larger ducts are concentrated centrally within each lobule and lie in a markedly vascular connective tissue. The secretory end-pieces, initially acinar in form, are lined by proacinar cells which exhibit intercellular canaliculi at the lateral cell membranes and a few dense granules in the apical cytoplasm. During the second phase of development extensive changes occur within the secretory end-pieces, which elongate to form a system of branching tubules. Component cells show an increased granular content, and those in the main body of the tubules differentiate into mucous cells. By 34 cm postnatum the proacinar cells in the bulbous endings of the tubules are replaced by special serous cells possessing intercellular canaliculi and secretory granules which are either electron-lucent or electron-dense. The sequence of changes that occur during postnatal development is discussed and related to possible functional activities. The early development of the ducts may be correlated with their role in homeostasis, while the later development of secretory tubules and the differentiation of secretory cell types may be related to the onset of weaning, and may possibly be induced by this major change in dietary habit.

Age Factors↗
PubMed1979

Effects of the tumor-promoting agent 12-O-tetradecanoylphorbol-13-acetate on normal and "preneoplastic" mouse submandibular gland epithelial cells in vitro.

The effect of the promoting agent 12-O-tetradecanoylphorbol-13-acetate (TPA) was studied on mixed primary cultures of C57BL/Icrf-at mouse submandibular gland and on slowly proliferating preneoplastic epithelial foci derived from such cultures. No cytologic alterations were noted in the TPA-treated epithelium. The development of three-dimensional epithelial ducts occurring between 30 and 60 days in vitro in untreated cultures was markedly inhibited in TPA-treated cultures. Epithelial proliferation, as shown by the [H]thymidine labelling index and mitotic index, seemded to increase during treatment with TPA and fall between the weekly treatments. Long-term TPA treatment increased the incidence of slow-growing foci in cultures previously treated with 7,12-dimethylbenz[a]anthracene but not in cultures treated with dimethyl sulfoxide. TPA had no effect on the emergence of tumorigenic cell lines at a late stage in culture or on the development of such lines from preexisting foci derived from non-TPA-treated cultures.

9,10-Dimethyl-1,2-benzanthracene↗