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PubMed2000

Rodent submandibular gland peptide hormones and other biologically active peptides.

The cervical sympathetic trunk-submandibular gland neuroendocrine axis plays an integral role in physiological adaptations and contributes to the maintenance of systemic homeostasis, particularly under the 'stress conditions' seen with tissue damage, inflammation, and aggressive behavior. The variety of polypeptides, whose release from acinar and ductal cells is under sympathetic nervous system control, offers coordinated and progressive levels of endocrine communication. Proteolytic enzymes (e.g. the kallikreins and furin maturases) are involved in the conversion of inactive precursors (e. g. Pro-EGF and SMR1) into biologically active molecules (e.g. EGF, SMR1-pentapeptide), which act on local or distant targets and thereby modulate the homeostatic process.

Animals↗

PubMed1991

Modulation of immunoreactive epidermal growth factor levels in the submandibular gland, pancreas, liver, kidney and gastrointestinal tract of suckling rats by cortisone and tri-iodothyronine.

Suckling rats exhibit age-dependent differences in epidermal growth factor (EGF) levels in several organs. The present studies evaluated the effects of two hormones known for their maturative effect on suckling rats, cortisone and tri-iodothyronine (T3), on immunoreactive EGF levels in specific organs. Suckling rats were administered cortisone (5 mg/100 g body weight per day) or T3 (50 micrograms/100 g body weight per day) on days 8, 9, 10 and 11 after birth, and killed on day 12. Submandibular glands, kidneys, pancreas, liver and gastrointestinal tract mucosa and lumen were assayed for immunoreactive EGF by a species-specific radioimmunoassay. Low levels of EGF in the submandibular glands were increased slightly by both T3 and cortisone treatment. Cortisone evoked a tenfold increase in EGF in the pancreas, but had no effect on levels in the kidney or liver. In contrast, T3 evoked a sixfold increase in the EGF level in the kidney, but had no effect on levels in the pancreas or liver. Hormonal administration had no effect on EGF levels in the stomach. Within the intestinal tract, cortisone had no effect on the luminal EGF content of the duodenum, jejunum or ileum, but caused a decrease in the midjejunum. T3 evoked a decrease in the luminal EGF content of the ileum. The effect of cortisone on mucosal EGF content varied between regions; an increase was seen in the duodenum with a decrease in the midjejunum and ileum. T3 administration resulted in a significant decrease in EGF only in the mucosa of the ileum.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1995

The extracts from rat submandibular glands induce the erythroid differentiation of K-562 cells.

The extracts from rat submandibular glands (SMGs) induced erythroid differentiation of K-562. The activity was non-dialysable and abolished by heat, trypsin or 2-mercaptoethanol. Follistatin, which neutralizes the erythroid differentiation factor (EDF), had no effects on this activity. Analysis by gel chromatography and polyacrylamide gel electrophoresis-isoelectric focussing showed that the characteristics of this substance were different from those of erythropoietin, TGF-beta 1, EDF, stem cell factor and insulin-like growth factor-1. These results suggest the presence of a novel substance in rat SMGs which induces erythroid differentiation of K-562.

Animals↗

PubMed1999

Postoperative irradiation in malignant tumors of submandibular gland.

A retrospective review of 18 patients with treated primary malignant tumors of submandibular gland is presented. The data revealed that initial control of malignancies was inversely related to the presence of disease and surgical margins. Postoperative radiation therapy was used in cases with positive surgical margins. Of the patients with inadequate surgical margins, 71% achieved local relapse when they had surgery alone, compared with 27% of the group that received postoperative irradiation. This study suggests that adjuvant radiation therapy may improve the local control and survival. Such therapy can serve as the salvage method for those who have inadequate surgical margins. Re-excision is disappointing in its ability to control this disease.

Adenocarcinoma↗

PubMed1968

Canine submandibular-gland hyaluronidase. Purification and properties.

1. Methods for the purification of dog submandibular-gland hyaluronidase from sedimentable and non-sedimentable portions of a homogenate and from the whole homogenate are presented. The method consists of three main steps: removal of mucin by acid precipitation or gel filtration on Sephadex G-200, ammonium sulphate precipitation and CM-cellulose chromatography. By this method specific activities of up to 1.28 and 0.78mumoles of N-acetylglucosamine/min./mg. of protein were obtained for the purified freeze-dried non-sedimentable hyaluronidase and for the sedimentable hyaluronidase respectively. 2. A comparison of some of the properties of the non-sedimentable and the sedimentable hyaluronidase preparation indicated that there was little difference between the two and that they both resembled lysosomal hyaluronidase from rat liver.

Animals↗

PubMed2003

Distribution of sympathetic and afferent neurones innervating the submandibular gland in the sheep.

The distribution of sympathetic and sensory neurones innervating the submandibular gland (SMG) in sheep was studied using retrograde tracing technique. The retrograde tracer Fast Blue (FB) was unilaterally injected into the SMG in five juvenile male sheep under general anaesthesia. After a 4-week survival period, all the animals were reanaesthetized, perfused transcardially with 4% buffered paraformaldehyde and ganglia, which could be considered as a potential sources of sympathetic, and afferent innervation of the gland were bilaterally collected. The FB-labelled sympathetic neurones were found in the ipsilateral superior and middle cervical ganglion. Many labelled neurones were distributed in the ipsilateral jugular and nodose ganglia of the vagus nerve and smaller numbers of the nerve cells were also found in ipsilateral C1-C3 dorsal root ganglia (DRG). No labelled neurones were observed in the ipsilateral stellate ganglion, trigeminal ganglion, C4-C8 DRG and in all contralateral ganglia. The present study revealed that the majority of sympathetic neurones projecting to the sheep SMG are found in the superior cervical ganglion but some of them are also distributed in the middle cervical ganglion. Most of the afferent neurones are located in the jugular and nodose ganglia of vagus nerve but C1-C3 DRG also comprise some of these nerve cells.

Animals↗

PubMed1983

Differences in the ultrastructure of the submandibular glands of baboon and Rhesus monkey revealed by the use of different fixatives.

Submandibular glands of adult baboon and Rhesus monkeys were compared after different methods of fixation. In both species, serous acinar cells outnumber mucous acinar cells. In the baboon, serous cells contain secretory granules showing dense cores, moderately dense crescents, and flocculent material. In mucous cells, secretory granules vary in appearance from amorphous to highly ordered depending on fixation. In the Rhesus monkey, serous cells contain the same 3 components of secretory granules as in the baboon. Additionally, a fourth component is represented by a layer of electron-dense material between the crescent and flocculent material. Mucous cells contain electron-dense granules when fixed in Millonig's buffered fixatives, but when Clarke's or Sorensens' buffer is used the granules resemble more typical mucous granules. Duct systems of the two species are similar, but differ mainly in that large foci of glycogen are present in the striated duct cells of the Rhesus monkey.

Animals↗

PubMed1980

Age related changes in NGF, EGF and protease in the granular convoluted tubules of the mouse submandibular gland. A morphological and immunocytochemical study.

Granular convoluted tubule (GCT) cells in the submandibular glands of male mice were studied at 1.5, 6, 12 and 29 months of age by morphometric and immunocytochemical means. The relative proportion of GCT's was established by determining a tubule to acini (T/A) ratio. The T/A ratios were significantly increased between 1.5 months (0.76) and 6 months (1.34), and between 6 months and 12 months (1.96). However, a marked decrease was found at 29 months (0.59). The functional capacity of GCT cells was estimated by immunocytochemical staining for nerve growth factor (NGF), epidermal growth factor (EGF), protease and renin. The number of GCT cells immunoreactive for NGF, EGF and protease increased between 1.5 months and 6 months. No further change was noted at 12 months. However, once again at 29 months a decrease occurred in a number of cells reactive for all three substances. Renin was not detected in any cell at any age.

Age Factors↗

PubMed1983

Prenatal and postnatal histogenesis of myoepithelium in hamster submandibular gland. An ultrastructural study.

Prenatal and postnatal development of myoepithelium in the hamster submandibular gland has been studied by electronmicroscopy. Myoepithelial cells originate from 'undifferentiated' stem cells in the epithelial anlage at 14-1/2 days of gestation. At this stage of their cytodifferentiation, the myoepithelial cells are recognizable by their elongated shape, peripheral location between the basal lamina and other epithelial cells in the anlage, axially oriented dense nucleus and low density cytoplasm. The myofilament deposition starts at birth, initially in the basal cytoplasm. With advancing maturation, there is a progressive increase in the number of myofilaments and in two week-old animals they virtually fill the cytoplasm. The mature myoepithelial cells are characterized by their shape, location, cytoplasmic myofilaments which run parallel to the basal lamina and display focal densities along their course, hemidesmosomes and pinocytotic vesicles. The morphologic evidence supports their epithelial origin but militates against their ability to mediate the synthesis of fibrous, mucinous, chondroid or osseous material as proposed by others (Grishman, 1952; Azzopardi and Smith, 1959; Doyle et al., 1968; Hubner et al., 1971).

Epithelium↗

PubMed1985

Histochemistry of hydrolytic enzymes in resting submandibular glands of rabbits.

The distribution of several hydrolytic enzymes was investigated in rabbit submandibular glands at both the light and electron microscopical levels. Glands were fixed by either immersion or perfusion fixation with a variety of fixatives containing 1-2% glutaraldehyde and 2-4% formaldehyde in 0.1 M cacodylate buffer at pH 7.2. Light microscopically, the acinar cells showed some staining for ATPase, acid phosphatase and nonspecific esterases but showed weak staining for thiamine pyrophosphatase. Acid phosphatase staining occurred most strongly in granular tubule cells. Staining for esteroproteases was confined to the periluminal rims of intercalary and striated ducts. Alkaline phosphatase was very sensitive to glutaraldehyde and was confined to myoepithelial cells. Electron microscopical observations revealed the presence of acid phosphatase reaction product in lysosomes, immature granules and in GERL-like structures, the last being much more conspicuous in the granular tubule cells. ATPase reaction product was localized to the basal and luminal plasma membranes and lumina of both acinar and granular tubule cells. The Golgi complex of these two types of cells exhibited only moderate amounts of reaction product for thiamine pyrophosphatase. Alkaline phosphatase activity, on the other hand, was exclusively localized to myoepithelial cells in their plasma membranes and sometimes in the nuclear envelope.

Acid Phosphatase↗

PubMed1989

Changes with senescence in the fine structure of the granular convoluted tubule of the submandibular gland of the mouse.

Cells of the granular convoluted tubules (GCTs) of the submandibular gland of senescent male mice show structural changes indicative of functional decline. In order to define the nature of these age-related changes more clearly, the fine structure of GCT cells of 12- and 28-month-old males was compared. In old mice, there was cell-to-cell variation in the extent of these changes, with some cells of senescent males appearing no different from those of young adults. In affected cells the most striking alterations were seen in secretion granules and lysosomal elements. Secretion granules varied greatly in size, with some GCT cells having only very fine apical granules. Secondary lysosomes and large lipofuscin granules were frequent in the basal cytoplasm. Very large dense bodies (3-5 micron) occurred in many cells. These possibly represent intracellular pools of released secretory materials, as they were occasionally seen in continuity with the luminal contents. Structures whose appearance was intermediate between the very large dense bodies and lipofuscin granules were common, suggesting crinophagic activity. There was an apparent decrease in numbers of polysomes and in the extent of the Golgi apparatus. These fine structural changes are consistent with impairments with advanced age in synthesis and posttranslational processing of secretory products by affected GCT cells. In addition to cell-to-cell variation in any one male, there was also interanimal variation in the degree and extent of these senescent changes.

Aging↗

PubMed1987

Regeneration of submandibular gland autografts in sympathectomized rats.

This morphologic study compares the regenerative response in submandibular gland (SMG) autografts placed in the tongues of previously sympathectomized rats to autografts placed in tongues of sham-sympathectomized rats. We hypothesized that sympathectomy would alter the process of cellular proliferation and inhibit cytodifferentiation in regenerating SMG autografts. Either 1 week, or 8 to 11 weeks following the SMG autografting procedure, the rats were sacrificed and their tongues were removed and sectioned in a cryostat. Frozen tissue sections containing the SMG autografts were either reacted for cholinesterase activity, treated with a glyoxylic acid mixture to induce histofluorescence, or stained for histologic examination. In addition, 3H-thymidine labeled and unlabeled cells were counted in autoradiographs of 1-week autografts, and these counts were used to calculate labeling indices. The 1-week SMG autografts from both the sympathectomized and the sham-sympathectomized rats were similar in histologic appearance, and neither group of autografts contained cholinesterase-positive or monoaminergic nerve fibers. The 8- to 11-week autografts from sympathectomized and sham-sympathectomized rats contained cholinesterase-positive fibers, but monoaminergic fibers were present in the autografts only from the sham-operated rats. Acinar cells were observed in one-third of the 8- to 11-week autografts of both the sympathectomized and sham-sympathectomized rats. This finding suggests that sympathectomy did not prelude cytodifferentiation in the autografts. The autoradiographic data revealed no statistically significant difference between the mean labeling indices of the 1-week autografts from the sympathectomized and sham-sympathectomized rats, which suggests that sympathectomy also did not alter the level of cellular proliferation in the autografts.

Animals↗

PubMed2002

Activation of phospholipase D-2 by P2X(7) agonists in rat submandibular gland acini.

Exogenous ATP stimulated phospholipase D (PLD), but not sphingomyelinase in rat submandibular gland (SMG) acini. PLD activation was dependent upon extracellular Ca(2+) and did not involve intracellular Ca(2+) mobilization or phosphoinositide-specific phospholipase C activation. ATP-stimulated PLD was attenuated by inhibition or downregulation of protein kinase C (PKC). PLD activation was fully blocked by the cytosolic phospholipase A(2) (PLA(2)) inhibitor ONO-RS-082 and partially attenuated by the selective Ca(2+)-dependent cytosolic PLA(2) inhibitor, arachidonyl trifluoromethylketone (AACOCF(3)), or by bromoenol lactone, an inhibitor of Ca(2+)-independent cytosolic PLA(2). Magnesium, which decreases the concentration of ATP(4-), and nickel, which blocks nonspecific cation channels coupled to purinergic receptors, inhibited PLD activation by ATP. Using reverse transcription-polymerase chain reaction and Northern blotting techniques, we demonstrated that the PLD isoform stimulated by ATP was PLD-2. Among various ATP analogs, only the P2Z/P2X(7) purinergic receptor agonist benzoyl-benzoyl ATP stimulated PLD-2. The response to ATP was inhibited by the nonselective P2X purinergic antagonist suramin and by oxidized ATP, a potent P2Z/P2X(7) receptor antagonist. It is concluded that in rat SMG acinar cells, PLD-2 is upregulated by exogenous ATP through a mechanism involving Ca(2+) influx, cytosolic PLA(2), and PKC. Also, the data suggest an involvement of P2X(7) receptors in PLD-2 stimulation by ATP.

Adenosine Triphosphate↗

PubMed1993

Characterization of growth and differentiation of normal human submandibular gland epithelial cells in a serum-free medium.

Parenchymal tissue of human submandibular glands was cultured in a serum-free medium consisting of a 1:9 mixture of Dulbecco's modified Eagle's medium and MCDB 153 supplemented with 10 ng/ml epidermal growth factor, 10 microM dexamethasone and 1 microgram/ml insulin. Cultivation of the tissue in this medium resulted in propagation of loosely arranged epithelioid cells on plastic, without the necessity of a matrix. Epidermal growth factor significantly enhanced mitogenesis of cultured cells, which expressed specific high- and low-affinity receptors for epidermal growth factor. The epithelioid cells were found to represent the undifferentiated ultrastructure of ductal cells. Immunocytochemically, cultured epithelioid cells expressed antigens specific to basal cells of the intra- and interlobular ducts in situ, including cytokeratins 3 and 6 and cytokeratins 13 and 16, vimentin, and alpha-smooth muscle actin. Moreover, cytoplasm of the cells was immunostained using antibody against the basement membrane component, type IV collagen. These results suggested that cultured epithelioid cells are undifferentiated ductal cells, which have the characteristics of basal cells of the intra- and/or interlobular ducts. Cultured epithelioid cells maintained the characteristics for serial passage until the time that the cultures were confluent. On the other hand, several stratified foci developed on the confluent monolayer. The stratified cells were strongly positive for cytokeratins 3 and 6, but negative for vimentin, alpha-smooth muscle actin and type IV collagen. Moreover, the stratified cells were strongly stained with the antibody against epithelial membrane antigen. This antibody stained the luminal membrane domain of salivary epithelial cells. Electron micrograph of the vertical section through the foci revealed stratified cell layers with a gradual transition from basal cells to squamous epidermoid cells. This result suggests that cultured epithelioid cells, which have the characteristics of basal cells of the intra- and/or interlobular duct, have the potential to differentiate into luminal duct cells.

Cell Differentiation↗

PubMedDate not supplied

Development of mouse submandibular gland studied by field emission scanning electron microscopy.

The ultrastructural organization of mouse submandibular gland from 15 days gestation to 180 days of age was studied by field emission scanning electron microscopy. At 15 days of gestation several groups of acini, intercalated and striated ducts were present. They consisted of numerous pyramidal and/or polyhedral cells with spherical or elongated nuclei and short microvilli. At 3 days after birth different shapes of acinar structures covered by a network of fine collagen bundles were observed. The acinar portions corresponded to the terminal tubules and showed the acinar cells containing immature secretory granules. Fractured specimens of ducts revealed a flat surface with large central nuclei. At 14 days after birth acinar terminal portions possessed a round shape; branches of either intercalated and striated ducts were also observed. Gap junctions and interdigitations were numerous at the base of acinar cells. At 30 days after birth the acinar terminal portions and striated ducts revealed a large number of secretory granules. Acinar cells showed a pyramidal shape and basal nuclei. Freeze-cracked surfaces of the striated ductal cells evidenced also a pyramidal shape. Secretory granules ranged from 0.3 to 1.2 microns in size and were clearly observed by infoldings of the basement membranes. At 90 days after birth the ultrastructural features were more differentiated when compared with the previous ages. The freeze-cracked specimens showed very numerous secretory granules in all acinar and striated ductal cells. The granules occupied almost all the apical region of the cells. The outer surface of the basement membrane of acinar and myoepithelial cells was constituted by a spongy-like material covered by fine collagen fibrils. At 180 days after birth numerous secretory granules were seen either in acinar and ductal cells. A morphofunctional polarization of the cell was finally clearly observed in that the cytoplasmic organelles were concentrated in the basal portion whereas the secretory granules were located in the apical region of the cells.

Animals↗

PubMed1991

Adrenergic interaction in mucin secretion of rat submandibular gland in vitro.

Previous studies have reported that maximally effective concentrations of the "mixed" alpha and beta adrenoceptor agonists, epinephrine and norepinephrine, cause greater amounts of mucin secretion than the "pure" beta adrenoceptor agonist, isoproterenol, and that this response requires extracellular calcium. The purpose of the present study was to examine directly the nature of the effect of the putative pure alpha adrenoceptor agonist, phenylephrine, on isoproterenol-induced mucin secretion and the role of extracellular calcium in this interaction. We used a graphical method which provides a procedure for determining whether interaction between two drugs is additive, antagonistic or synergistic to analyze this interaction. Submandibular glands were removed from male rats, divided into sections and placed in modified Hank's balanced salt solution. Mucin secretion was measured as acid precipitable disintegrations per minute after labeling the submandibular gland with [14C]glucosamine. Phenylephrine caused a small but significant secretion of mucin which was blocked partially by phentolamine and completely by propranolol. Phenylephrine caused a significant shift of the isoproterenol concentration-response curve to the left of the theoretical curve expected if two drugs act additively. Mucin secretion induced by isoproterenol alone was independent of extracellular calcium concentration; however, the combination of isoproterenol and phenylephrine caused a concentration-dependency on extracellular calcium.

Animals↗

PubMed1996

Ancient schwannoma of the submandibular gland: a case report.

Schwannomas (neurilemmomas) are neurogenic tumors that arise from the Schwann cells of the neural sheath. They are most often benign and solitary. Extracranial schwannomas are rare, and can be mistaken for metastatic disease or other non-neurogenic tumors. Ancient schwannoma is a rare variant of schwannoma with a course typical of a slow-growing benign tumor. Histologically, it can be confused with a malignant mesenchymal tumor. An unusual case of an ancient schwannoma of the submandibular gland is reported. The clinical, histological and surgical aspects of this tumor are discussed, and the literature regarding this rare entity is reviewed.

Adult↗

PubMed1999

Salivary secretion of highly concentrated chromogranin a in response to noradrenaline and acetylcholine in isolated and perfused rat submandibular glands.

Chromogranin A (CgA) is a member of a family of highly acidic proteins, chromogranins, which are co-stored in the adrenergic neurons and paraneurons and co-released with adrenaline and noradrenaline (NAd) in response to adequate stimulation. The present study provides novel evidence that CgA-like immunoreactivity (IR) is stored in the exocrine cells in the granular convoluted tubule, and is secreted into saliva by stimulation with NAd and acetylcholine (ACh) in the isolated and perfused rat submandibular gland. NAd at 1 microM produced maximum secretion of CgA-like IR (<< 0.9 mM) and a marked increase in salivary flow. Further increases in NAd concentration (10 or 100 microM) yielded concentration-dependent decreases in both responses. ACh at 1 microM produced maximum salivary flow and a slight elevation of CgA-like IR secretion (6 microM); 100 microM ACh decreased the salivary flow but increased the CgA-like IR secretion (0.6 mM). Electron microscopic examination showed vigorous compound exocytosis of secretory granules in the cells of the granular convoluted tubule when the submandibular gland was stimulated with 1 microM NAd. These results provide an experimental basis for the view that the salivary CgA-like IR secretion may be a sensitive and quantitative index of the activity of the sympathetic nervous system innervating the gland.

Acetylcholine↗