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PubMed1989

Gustatory-salivary reflex: neural activity of sympathetic and parasympathetic fibers innervating the submandibular gland of the hamster.

Electrophysiological experiments were performed to clarify the neural control mechanisms subserving gustatory-salivary reflex in anesthetized and decerebrate hamsters. Efferent neural activities of postganglionic sympathetic and preganglionic parasympathetic fibers, innervating the submandibular gland, were recorded when taste stimuli were infused into the oral cavity. Neural activities of primary gustatory afferents were also recorded from the chorda tympani (innervating the anterior part of the tongue) and the glossopharyngeal nerve (innervating the posterior part of the tongue). The parasympathetic fibers showed a low rate of spontaneous discharges (about 0.3 Hz), and responded tonically in an excitatory manner to taste stimulation. The magnitude of parasympathetic activity was highly correlated with the magnitude of gustatory afferent responses of the chorda tympani rather than that of the glossopharyngeal nerve. On the other hand, the sympathetic fibers showed irregular burst discharges (1.5 burst/s), and the rate of burst discharges was increased in response to high concentrations of HCl (0.03 M) or NaCl (1 M) solutions. Deafferentation experiments suggest that the parasympathetic activity is mainly influenced by gustatory information via the chorda tympani, while the sympathetic activity can be evoked by both the chorda tympani and glossopharyngeal nerve.

Action Potentials↗
PubMed2005

Clock gene expression in the submandibular glands.

Clock genes, which mediate molecular circadian rhythms, are expressed in a circadian fashion in the suprachiasmatic nucleus and in various peripheral tissues. To establish a molecular basis for circadian regulation in the salivary glands, we examined expression profiles of clock-related genes and salivary gland-characteristic genes. Clock-related genes-including Per1, Per2, Cry1, Bmal1, Dec1, Dec2, Dbp, and Reverbalpha-showed robust circadian expression rhythms in the submandibular glands in 12:12-hour light-dark conditions. In addition, a robust circadian rhythm was observed in amylase 1 mRNA levels, whereas the expression of other salivary-gland-characteristic genes examined was not rhythmic. The Clock mutation resulted in increased or decreased mRNA levels of Per2, Bmal1, Dec1, Dec2, and Dbp, and in Cry1-/- background, Cry2 disruption also increased or decreased mRNA levels of these clock-related genes and the amylase 1 gene. These findings indicate that the Clock- and Cry-dependent molecular clock system is active in the salivary glands.

ARNTL Transcription Factors↗
PubMed1987

Genes encoding proteins with homologous contiguous repeat sequences are highly expressed in the serous cells of the rat submandibular gland.

An abundant class of secreted salivary polypeptides is characterized by the presence of identical and contiguous repeats of amino acid sequences within the polypeptide chains, and includes the proline-rich proteins. We discovered a new family of contiguous repeat polypeptides (CRPs) that is related to the proline-rich proteins but contains little proline. Analysis of salivary mRNAs and liver DNA by molecular cloning, DNA sequence determinations, and Northern and Southern blot hybridization revealed several closely related CRP mRNAs and at least 10 CRP-related genes. We further analyzed two CRP mRNAs of 850 and 920 nucleotides and the gene encoding the larger CRP mRNA. The two mRNAs contain the same 69-base repeats in their coding regions and are identical in their 5'- and 3'-untranslated tracts. However, they differ in the number of contiguous repeats (four versus five) and a segment at the 3' end of the coding region which encodes closely related but unique COOH termini of the CRPs. These structural features suggest a recent gene conversion. The CRP gene analyzed is divided into three exons that encode (i) 5'-untranslated tract and signal sequence, (ii) secreted polypeptide, and (iii) 3'-untranslated tract, respectively. CRP mRNA contains two open reading frames. The longer open reading frame encodes a CRP precursor with a signal sequence of 17 amino acids, four to five contiguous repeats of 23 amino acids, and a variable COOH region that begins with two segments related to the contiguous repeats. Immunochemical analysis of salivary gland slices with antisera raised against peptides corresponding to two regions of the larger open reading frame revealed intense staining only of the serous cells of the submandibular glands. 35S-Labeled oligonucleotides complementary to CRP mRNA specifically hybridized to the same cells.

Amino Acid Sequence↗
PubMed2006

Cytosolic calcium oscillations in submandibular gland cells.

Calcium oscillations can, by default, encode diverse and specific signals by different modes of modulation. Frequency modulation is illustrated by the activation of calcium/calmodulin-dependent protein kinase II at unit Hz, and of calcineurin at 10 mHz frequencies, respectively. The submandibular gland secretory axis is characterized by both potassium and osmolarity gradients from the luminal side of the secretory cells. Such gradients may play significant physiological roles through the feedback modulation of cholinergic stimulation. High potassium transforms plateau calcium increases induced by cholinergic stimulation of the submandibular acinar cells into oscillatory calcium increases. The ductal cells may have similar mechanisms of feedback modulation both by high potassium and by hypoosmolarity. Such feedback mechanisms could modulate the decision-making process for determining which secretory products are selectively released after nerve stimulation.

Animals↗
PubMedDate not supplied

[Oscillatory changes in cells of the submandibular glands in mice during the secretory cycle].

By means of 3H-leucine radioautography, ultrastructural and morphometrical analysis, it has been demonstrated that in the cells of the acinar (Ac) and granular (Gr) parts of the submandibular gland (SMG) in mice with a synchronized for 3 h nutrition cycle there are endogenic fluctuations (for about 1 h) of the secretory process. They are demonstrated as alterations in intensity of protein synthesis in the cells, their areas and ultrastructure. In the Ac cells at the moment of feeding and in 50, 110, 150 min after feeding maximal leucine incorporation is observed. The changes in the cell area during the first hour occurs with 10 minutes' overtake of the incorporation intensity, and during the following 2 h an equilibrium is reached between these two parameters, when the maximal contents of the label corresponds to the minimal area of the cells. In the Gr cells one maximum of 3H-leucine incorporation occurs in 60-80 min after feeding. The minimal content of the label takes place in 10 and 120 min after feeding. During the time mentioned inverse ratio between the intensity incorporation of the labelled isotope and the change of the cell volume is observed. This is connected with autoregulatory processes in the cells. It is possible that in the Ac cells together with removal of the secretory granules by means of exocytosis (in 10-20 min after feeding) the secretory material is removed before it is formed as granules by parvivesicular extrusion. In the Gr cells apocrinic and endocrinic secretion takes place. Presence of certain specific formations (secretory lysosomes) explains peculiarities of the secretory process in the SMG.

Animals↗
PubMed2004

Additive and/or synergistic action (downregulation) of androgens and thyroid hormones on the cellular distribution and localization of a true tissue kallikrein, mK1, in the mouse submandibular gland.

We investigated the effects of 5alpha-dihydrotestosterone (DHT), 3,5,3'-triiodo-l-thyronine (T(3)), and dexamethasone (Dex) on the expression of mK1 in the granular convoluted tubule (GCT) cells of the submandibular gland (SMG) of hypophysectomized (Hypox) male mice by indirect enzyme-labeled antibody and immunogold antibody methods for light and electron microscopy. Hypox resulted in considerable atrophy of the GCT cells, which were always immunoreactive for mK1, and the cells were characterized by apical small dense secretory granules labeled with gold particles suggesting the presence of mK1, small Golgi apparatus, sparse rough endoplasmic reticulum (RER), and developed basal infoldings. Each of the hormones, DHT, T(3), and Dex, enhanced the GCT phenotype to various degrees in Hypox male mice. Both DHT alone and T(3) alone moderately inhibited mK1 synthesis by increasing the number of mK1-immunonegative GCT cells in Hypox males, but Dex alone had no inhibitory effect on mK1 synthesis. A significant trophic effect on GCT cells was induced by combined injection of DHT and T(3) or of all three hormones, and was reflected in the appearance of abundant large secretory granules, well-developed Golgi apparatus and RER, and reduced basal infoldings. Only a few such GCT cells were immunopositive for mK1, and the pattern of immunopositive and immunonegative cells very closely resembled the mosaic pattern seen in normal male GCTs. These findings suggested that the sexual dimorphism of mK1 expression and the morphological appearance of GCT cells can be induced by treatment with two hormones, DHT and T(3), but not by either of them alone. T(3) appears to have a permissive effect on committed GCT cells that results in downregulation of mK1 expression in these cells.

Androgens↗
PubMed1981

Postsynaptic localization of alpha 2-adrenergic receptors in rat submandibular gland.

Norepinephrine is known to inhibit its own release from presynaptic nerve terminals through alpha 2-adrenergic receptors, which presumably have a presynaptic localization. alpha 2-Adrenergic receptors (as determined by [3H]clonidine binding) appear in rat submandibular gland membranes following reserpine treatment. These alpha 2 receptors seem to be localized postsynaptically, based on the following evidence. (1) Partial destruction of the presynaptic nerve terminals with 6-hydroxydopamine did not decrease the density of alpha 2-adrenergic receptors following subsequent reserpine administration. (2) Duct ligation, which results in atrophy of the gland, markedly decreased the density of the receptors following subsequent reserpine administration. (3) Surgical denervation resulted in the appearance of high levels of alpha 2-adrenergic receptors. (4) The changes in alpha 2 receptors paralleled the changes in postsynaptic beta-adrenergic receptor binding (as determined by [3H]dihydroalprenolol). While these results establish the existence of postsynaptic alpha 2-adrenergic receptors in an innervated tissue, the concomitant presence of a low density of presynaptic alpha 2 receptors has not been eliminated.

Animals↗
PubMedDate not supplied

Inhibition of branching morphogenesis of mouse fetal submandibular gland by sodium fluoride--protection by epidermal growth factor.

As an initial step to study the effect of sodium fluoride on the oral environment, we investigated how sodium fluoride (NaF) affects the branching morphogenesis of fetal mouse submandibular gland (SMG). When mouse SMG was prepared from the embryo at 13 days post prenatal stage and cultured, gradual development of branching morphogenesis was observed. Addition of NaF affected this morphological change in bimodal fashions. At a lower concentration of NaF (< 2 microM), the branching morphogenesis was slightly enhanced, whereas at a higher concentration of NaF (4 - 8 microM), it was almost completely inhibited. The inhibitory effect of NaF at the higher concentration was abrogated by stimultaneous addition of epidermal growth factor (EGF), but not by 5alpha-dihydrotestosterone (DHT) or insulin-like growth factor (IGF). These data demonstrate that EGF can effectively reduce the cytotoxic activity of NaF at micromolar concentration.

Animals↗
PubMed1977

Human submandibular gland tissue in culture 2. Nickel affinity to secretory proteins.

In view of the exposure to nickel through the oral cavity the relation between this metal (as 63NiCl2) and salivary macromolecules was studied by tissue culture, isoelectric focusing and equilibrium dialysis. During cultivation of human submandibular gland tissue, small amounts of nickel were incorporated into macromolecules with an isoelectric point of about 4.4 and approximately 7.5. No affinity independent of the protein synthesis was revealed between nickel and submandibular proteins, including the acidic, viscous, sulphate-incorporating glycoproteins. It is therefore possible that a nickel-containing metalloprotein is present in the submandibular secretion, but a complex formation between nickel and macromolecules similar to the nickel-albumin transport system in plasma was not found. The affinity of nickel to small molecular substances in the secretion was not studied.

Culture Techniques↗
PubMed1992

Sexually dimorphic duct system of the submandibular gland in mouse with testicular feminization mutation (Tfm/Y).

The X-linked testicular feminization mutation (Tfm/Y) in the mouse is characterized by androgen insensitivity of the target cells. The aim of this study was to examine sexually dimorphic development of the submandibular gland of Tfm/Y mutant mice in comparison with those of wild-type male, wild-type female and heterozygous Tfm female mice. In either 30- or 90-day-old wild-type male mice, the granular convoluted tubules (GCT) of the glands were more developed, and the relative occupied areas (ROA) of GCT were superior to those of the age-matched wild-type and heterozygous Tfm females. In androgen-insensitive Tfm/Y mice, the glandular structures rather resembled the female glands, showing lower values of the ROA of the GCT. Sex differences in the mitotic rate were observed at 30 days of age, being significantly higher in the wild-type male GCT than in the female GCT. Thereafter, the mitotic rate of the wild-type male GCT declined to the female levels by 90 days of age. The mitotic rate of GCT in Tfm/Y mutants was as low as those of the females during observation periods. An other three regions, the acini, the intercalated ducts and the excretory striated ducts, were not significantly different in either the ROA or the mitotic rate among wild-type males and females, and Tfm/Y. On the other hand, either the ROA or the mitotic activity of GCT of the glands in Tfm/Y mutants was completely unaffected by 5 alpha-dihydrotestosterone (DHT).(ABSTRACT TRUNCATED AT 250 WORDS)

Androgen-Insensitivity Syndrome↗
PubMed2005

Lipopolysaccharide-induced elevation and secretion of interleukin-1beta in the submandibular gland of male mice.

The intraperitoneal injection of lipopolysaccharide (LPS) (400 microg/kg body weight) induced the expression of mRNAs of inflammatory cytokines such as interleukin (IL)-1beta, IL-6 and tumour necrosis factor (TNF)-alpha in the submandibular gland (SMG) of C3H/HeN mice but not that of C3H/HeJ mice, a mutant strain for Toll-like receptor-4 (TLR-4(-) mutant). The mRNA levels of these cytokines in the SMG of the wild-type mice increased as early as 3 hr after injection, peaked at 3-6 hr, and had decreased again by 24 hr. In this study, we particularly focused on IL-1beta, and induction by this endotoxin was investigated in detail. Denervation of the superior cervical trunk and chorda tympani nerve did not diminish the LPS-induced elevation of IL-1beta mRNA in the SMG, indicating the irrelevance of the central nervous system in this induction. TLR-4 mRNA and protein were shown to be strongly expressed in the SMG, suggesting the direct action of LPS on this gland. IL-1beta proteins were localized in the secretory granules of granular convoluted tubular (GCT) cells, and their molecular weights in the gland were 17.5 and 20 kDa. IL-1beta of the same size appeared in the saliva 6 hr after LPS injection in C3H/HeN but not in C3H/HeJ mice. The present study thus suggests that IL-1beta, an inflammation cytokine, is induced and secreted into the saliva in response to endotoxin injected intraperitoneally.

Animals↗
PubMed2001

Involvement of heparin-binding EGF-like growth factor and its processing by metalloproteinases in early epithelial morphogenesis of the submandibular gland.

In the present study, the role of a member of the epidermal growth factor (EGF) family, heparin-binding EGF-like growth factor (HB-EGF), in organ development was investigated by using developing mouse submandibular gland (SMG), in which the EGF receptor signaling and heparan sulfate chains have been implicated. HB-EGF mRNA was detected in developing SMG by RT-PCR analysis and was expressed mainly in epithelium and weakly in mesenchyme of the embryonic SMG. Epithelial morphogenesis was inhibited by a synthetic peptide corresponding to the heparin-binding domain of HB-EGF and by anti-HB-EGF neutralizing antibody. An in vitro assay using an EGF receptor ligand-dependent cell line, EP170.7 cells, allowed us to detect the growth factor activity in SMG-conditioned media, which was significantly reduced by anti-HB-EGF antibody. Furthermore, treatment of SMG rudiments with the hydroxamate-based metalloproteinase inhibitor OSU8-1, which inhibits processing of EGFR ligands including HB-EGF, markedly diminished the growth factor activity in conditioned media and resulted in almost complete inhibition of SMG morphogenesis. The inhibitory effects on morphogenesis were reversed, though partially, by adding the soluble form of HB-EGF. Our results provide the first evidence that HB-EGF is a crucial regulator of epithelial morphogenesis during organ development, highlighting the importance of its processing by metalloproteinases.

Amino Acid Sequence↗
PubMed2005

Morphological alterations induced by cytochalasin D on serous cells of human submandibular gland in basal and stimulated conditions.

Cytochalasin D (CD) is a fungal toxin which binds to the faster growing end of actin microfilament and inhibits actin polymerization. By an in vitro incubation system of slices of human submandibular glands obtained at surgery, we investigated by light microscope (LM), transmission electron microscope (TEM), and high resolution scanning electron microscope (HRSEM) the morphological changes caused by CD on serous cells. We studied the effects of the drug on secretory events induced by isoproterenol (I) and carbachol (C). With LM, following CD incubation, canaliculi were enlarged and prominent vacuoles were seen throughout the cytoplasm. By TEM, the vacuoles, which in many cases were in continuity with the lumen, represented the distinctive feature of secretory cells. With HRSEM, intercellular canaliculi, seen from their cytoplasmic side, exhibited many small spherical bulges, corresponding to the coated pits seen with TEM and indicating that the retrieval of plasma membrane was arrested at an early phase by the disruption of the actin cytoskeleton. In specimens treated with secretagogues and CD, a consequence reported here for the first time was the presence of dense granules within the vacuoles. The protrusions seen by HRSEM on the cytoplasmic side of intercellular canaliculi, following secretagogues stimulations, appeared peculiar to each stimulants, even if combined with CD, suggesting that besides actin filaments, other components, unaffected by CD, also are involved in the process of exocytosis and related phenomena.

Actin Cytoskeleton↗
PubMed2003

Apoptosis and apoptosis-related molecules in the submandibular gland of the nonobese diabetic mouse model for Sjögren's syndrome: limited role for apoptosis in the development of sialoadenitis.

Sjögren's syndrome is an autoimmune disease in which lymphocytic infiltrates develop in the exocrine glands. Pathogenetic aspects of the disease can be studied in the nonobese diabetic (NOD) mouse strain, a spontaneous model for Sjögren's syndrome. Apoptosis may play a role in the initation phase and in the effector phase of autoimmune diseases. Here, we have examined the role of apoptosis in the development of sialoadenitis in the NOD mouse. Apoptotic cells and the expression of apoptosis-related molecules were studied in submandibular glands (SMG) of NOD and NOD-scid mice before and after the onset of sialoadenitis. Numbers of apoptotic cells were not increased as compared with control mice, at any age. By immunohistochemistry, we demonstrated increased expression of Fas, Fas ligand (FasL), and bcl-2 on SMG epithelial cells of NOD and NOD-scid mice, as early as 3 days of age. mRNA expression of Fas and FasL was also examined in SMG by RQ-PCR. Low-level expression of Fas and FasL mRNA was observed in all mouse strains, from 1 day of age onward. We conclude that increased protein expression of Fas and FasL on SMG epithelial cells of NOD and NOD-scid mice probably indicates a genetically programmed abnormality in these cells that may form a trigger for the development of sialoadenitis in NOD mice. Because increased numbers of apoptotic cells were not observed, a role for actual apoptosis in the initiation or effector phase of sialoadenitis in the NOD mouse is unlikely.

Animals↗
PubMed1991

The role of substance P in parasympathetic nerve-induced secretion in the rat submandibular gland.

Fluid secretion from rat submandibular saliva evoked by electrical stimulation of the chorda tympani (parasympathetic secretory stimulation) was greatly reduced in the presence of atropine. However, some secretion was always evident at frequencies of 5 Hz and higher. The substance P-antagonist ([D-Arg1, D-Pro2, D-Trp7,9, Leu11]-substance P) also reduced the chorda-evoked saliva. Salivary secretion induced by the electrical stimulation of the superior cervical ganglion (sympathetic secretory nerve) was not affected by either atropine or substance P-antagonist. Continuous chordal stimulation reduced the glandular content of substance P to approximately 50% after 60 min, but similar stimulation of the superior cervical ganglion failed to produce such a reduction. Electron histochemical observation showed that the chordal stimulation caused depletion of substance P from the appropriate nerve fibers within the gland. The reduced secretory capability after continuous stimulation of the chorda at high frequency (20 Hz) was reversed by infusion of a subthreshold dose of exogenous substance P. Histological examination of granular duct cells revealed no degranulation in either chorda-stimulated or substance P-infused rats. The summation of findings suggests that endogenous substance P plays a complementary role in the regulation of parasympathetic nerve-induced fluid secretion in the acinus but is minimally involved in degranulation from granular duct cells.

Animals↗
PubMed1986

Unique radiosensitivity of serous cells in rhesus monkey submandibular glands.

The saliva of patients undergoing radiation therapy for head and neck cancer contains increased acidic mucosubstances associated with a reduced serous component. To assess the morphologic features of the acute radiation damage in serous versus mucous acinar cells, the mixed serous/mucous submandibular glands of 18 rhesus monkeys were studied 1-72 hours after irradiation with single doses of 2.5-15.0 Gy. Selective degeneration and necrosis of serous cells was observed with doses of 2.5-7.5 Gy. Doses of 10.0-15.0 Gy caused widespread destruction of whole serous acini, but only isolated mucous cells were affected. The lesions were clearly expressed by 24 hours. Transient exudation of neutrophils was replaced by plasma cells and lymphocytes. Examination at 16, 22 and at 40 weeks revealed that late atrophy was caused solely by loss of serous acini in glands treated with 7.5 and 10.0 Gy. Although both serous and mucous acini were reduced in glands treated with 12.5 and 15.0 Gy, the atrophy was mainly due to loss of serous acini. The finding that serous cells are more vulnerable to radiation injury than mucous cells provides a morphologic explanation for early and late changes in saliva composition after salivary gland irradiation.

Animals↗