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PubMed1997

Species differences in the expression of transforming growth factor-alpha (TGF-alpha) in the submandibular gland and pancreas.

CONCLUSION: Significant differences exist in the expression of transforming growth factor-alpha (TGF-alpha) in the submandibular glands (SMG) and the pancreas of different species and among cell components in the same species. BACKGROUND: Our previous studies have shown marked differences in the expression of TGF-alpha in the pancreas of humans and Syrian hamsters. To examine whether these differences also exist in other species, we examined the expression of TGF-alpha in the pancreas of mouse, rat, Syrian hamster, guinea pig, rabbit, pig, dog, and monkey. We included the SMG of these species for comparison. MATERIALS AND METHODS: The formalin-fixed tissues of these species (n = 3) were investigated by immuno-histochemistry using a monoclonal antibody to TGF-alpha. The SMG of rat, mouse, hamster, rabbit, pig, dog, and monkey were examined by RT-PCR to assure the specificity of the antibody. RESULTS: Remarkable species differences were found in the expression of this peptide in both the SMG and the pancreas. In the SMG, the expression varied in different cell components, even in the same tissue of the species. Although excretory and secretory ducts of the SMG of most species reacted with the antibody, intercalated ducts were immunoreactive only in mouse and guinea pig. Acinar cells were either weakly positive or nonimmunoreactive. In the pancreas of most species, the cells of the large and medium-sized ducts expressed TGF-alpha, whereas centroacinar cells of only rat and dog reacted with the antibody. Marked differences were found in the expression of TGF-alpha in islet cells and in its spatial distribution. Differences were also found in the immunoreactivity of mesenchymal and neural cells among the species.

Animals↗
PubMed1987

Effects of dopamine on the secretion of glycoproteins from the functional segments of the rat submandibular gland.

The action of dopamine (DA) on salivation and the secretion of marker glycoproteins (GP) from secretory cells of the rat submandibular gland (SMG) was investigated using various blockers at doses of 1 or 2 mg/kg (i.v.). DA at doses from 5 to 40 mg/kg (i.p.) dose-dependently increased salivation and the concentration of protein in SMG saliva. The order of inhibitory potency on salivation was propranolol (PPR) greater than phentolamine (PHN) much greater than haloperidol (HAL) when DA was administered i.p. at a dose of 10 mg/kg and PHN much greater than HAL much greater than PPR when the dose of DA was 40 mg/kg. The concentration of protein in saliva after pretreatment with HAL or PHN increased significantly at a dose of 40 mg/kg of DA, but did not increase at a dose of 10 mg/kg of DA. Moreover, pretreatment with PPR decreased it at both doses of DA. The electrophoretic profiles of GP in DA-evoked saliva showed two characteristic main bands of GP I (130 KDa) and GP IV (21.5 KDa) contained in the acinar cells (AC) and a minor band of GP III (31 KDa) which originated from the granular tubular cells (GT). The profile was not changed by pretreatment with PHN and HAL when DA was administered at a dose of 10 mg/kg, but at a dose of 40 mg/kg, the intensity of band I increased. Pretreatment with PPR, when DA was administered at 40 mg/kg, caused an increase in the intensity of band III and a reduction in that of band I. These results suggest that DA, at low doses, affects the AC, whereas at a higher dose, it affects both the AC and GT.

Animals↗
PubMed1989

[Biochemical characteristics of nuclear 5 alpha-reductase and cytosol 3 alpha-hydroxysteroid dehydrogenase in rat submandibular gland].

Biochemical studies of testosterone 5 alpha-reductase and 3 alpha-hydroxysteroid dehydrogenase (3 alpha-HSD) in rat submandibular gland (SMG) were performed. 14C-labeled testosterone or dihydrotestosterone (DHT) was incubated with subcellular fractions from rat SMG in the presence of 0.2 mM NADPH at 37 degrees C for 20 min in an atmosphere of 95% O2 and 5% CO2. Among the subcellular fractions, the high 5 alpha-reductase activity was detected in the nuclear fraction and 3 alpha-HSD in cytosol. Nuclear 5 alpha-reductase was efficiently solubilized in 2 mg digitonin per mg protein and 0.3 M KCl solution at 4 degrees C for 30 min. The maximum velocities (Vmax) of nuclear and solubilized 5 alpha-reductase activity for testosterone were 71.4 pmol/mg protein per min and 25.4 pmol/mg protein per min. Apparent Michaelis constant (Km) of nuclear and solubilized enzymes for testosterone were calculated as 11.1 microM and 16.7 microM by the Lineweaver Burk plot, respectively. The activity of solubilized 5 alpha-reductase from nuclei was stable by NADPH and KCl, and the molecular weight of the enzyme was estimated as 158 K.D approximately 200 K.D by Bio-Gel A-1.5 m column chromatography. The column chromatography also showed a peak of 3 alpha-HSD activity in cytosol, revealing the molecular weight of approximately 50 K.D. However, the elution peak of the 3 alpha-HSD was effectively decreased by KCl in Tris-HCl buffer. The molecular weight of 5 alpha-reductase and 3 alpha-HSD in SMG were similar to those in prostate. A stable and extractable 5 alpha-reductase was demonstrated in nuclei of rat SMG with possessing a considerable affinity for testosterone and also high 3 alpha-HSD activity for DHT was revealed in cytosol of the tissue.

3-Hydroxysteroid Dehydrogenases↗
PubMed1989

Study on high molecular weight androgen binding protein in submandibular gland of rat.

Testosterone (T) and 5 alpha-dihydrotestosterone (DHT) binding to cytosol and nuclear extract were investigated in the submandibular gland (SMG) and ventral prostate (VP) of rat at 3 and 7 weeks of age. The concentration of T and DHT were also measured by a specific radioimmunoassay (RIA) in the cytosol and nuclei of the tissues. The androgen binding protein analyzed by Sephacryl S-200 column chromatography showed a single binding peak in the nuclei and two critical peaks in the cytosol at 7 weeks old SMG and VP. The binding proteins were recovered predominantly as the mol wt of 150-200 kDa and 14-20 kDa in the cytosol and as a mol wt of 14-20 kDa in the nuclei. The two kinds of androgen binding protein were also revealed in the cytosol of 3 weeks old rat SMG. However, the S-200 column chromatography did not reveal the 150-200 kDa protein in the prostatic cytosol from 3 weeks rat, showing one binding peak of 14-20 kDa protein. The 150-200 kDa protein was specific to T in SMG at 3rd week and to DHT at 7th week in VP. The amount of specific binding of DHT to the 150-200 kDa protein was the greatest in VP cytosol of 7th week. The specific binding to the nuclear protein was shown by T in SMG at 3rd week and by DHT in VP at 7th week. High androgen concentrations were observed in the nuclei of male SMG (T: 3.92 +/- 0.72 ng/g, DHT: 4.14 +/- 0.51 ng/g) and the values were higher than in the nuclei of VP (T: 2.69 +/- 0.70 ng/g, DHT: 2.85 +/- 0.67 ng/g) at 3 weeks old. The values in SMG decreased to the levels of 1.67 +/- 0.68 ng/g for T and 1.19 +/- 0.09 ng/g for DHT at 7 weeks. The accumulation of T and DHT in the nuclei of VP were shown at 7 weeks old, revealing 4.29 +/- 3.18 ng/g for T and 5.53 +/- 2.39 ng/g for DHT. The cytosolic androgen levels in VP rather decreased from the 3rd week (T: 2.78 +/- 1.20 ng/g, DHT: 3.57 +/- 2.31 ng/g) to the 7th week (T: 1.33 +/- 0.40 ng/g, DHT: 1.80 +/- 0.66 ng/g). The T and DHT values in male cytosol and female cytosol and nuclei of SMG did not show a distinct increase or decrease with age.(ABSTRACT TRUNCATED AT 400 WORDS)

Androgen-Binding Protein↗
PubMed1977

Movement of horseradish peroxidase in submandibular glands of dogs after ductal injection.

Small volumes of solutions of a tracer enzyme, horseradish peroxidase (HRP), have been injected retrogradely via the main duct of submandibular glands in dogs, and the distribution of the peroxidase reaction product within the glands has been studied light microscopically and electron microscopically at different times after the injection. Tissue taken immediately after injections of 0.25 ml, given over 10 sec, showed that HRP had penetrated into the interstitial spaces of the gland. It was usually irregularly distributed and appeared to have reached the interstices of the gland by passing through the intercellular junctions of adjacent acinar cells. In time the tracer gradually disappeared from the interstices of the gland. Most of this removal was probably via lymphatic and blood vessels, into both of which peroxidase rapidly passed after ductal injection. At 5 hours after the HRP injection only very weak interstitial staining was present and at later times none was detected. An inflammatory reaction occurred in the gland, starting within minutes after the injection of this foreign protein and reaching a peak after about 12 hours. The events in dogs and those occurring in rabbits under similar experimental conditions have been compared. This study has given some indication how foreign substances may be distributed after retrograde injection into salivary glands.

Animals↗
PubMed2006

Expression and functional analysis of beta-adrenoceptor subtypes in rabbit submandibular gland.

beta-Adrenoceptors (beta-ARs) mediate important physiological functions in salivary glands. Here we investigated the expression and function of beta-AR subtypes in rabbit submandibular gland (SMG). Both beta(1)- and beta(2)-ARs, but not beta(3)-AR, were strongly expressed in rabbit SMG. beta(1)-AR proteins were widely expressed in acinar and ductal cells whereas beta(2)-AR proteins were mainly detected in ductal cells. A [(3)H]-dihydroalprenolol binding assay revealed that beta-AR B(max) was 186+/-11.9 fmol/mg protein and K(d) was 2.71+/-0.23 nM. A competitive binding assay with CGP 20712A, a beta(1)-AR antagonist, indicated that the proportion of beta(1)-AR and beta(2)-AR was 71.9% and 28.1%, respectively. Gland perfusion with the beta-AR agonist isoproterenol induced a significant increase in saliva secretion which was abolished by pretreatment with the non-selective beta-AR antagonist propranolol. Pretreatment with beta(1)- or beta(2)-AR selective antagonists, CGP 20712A or ICI 118551, diminished isoproterenol-induced increase in saliva secretion by 71.2% and 28.8%, respectively. The expression of alpha-amylase mRNA was significantly stimulated by isoproterenol, which was eliminated by propranolol and CGP 20712A. Perfusion with isoproterenol decreased alpha-amylase protein storage in SMG and increased alpha-amylase activity in saliva. These alterations became less significant after pretreatment with propranolol and CGP 20712A. Our results suggest that both beta(1)- and beta(2)-ARs are expressed in rabbit SMG. beta(1)-AR is the predominant subtype and may play an important role in regulating saliva and alpha-amylase secretion.

Animals↗
PubMed2000

Intraductal papilloma of the submandibular gland.

Salivary tissue intraductal papillomas are rare, benign tumours that predominantly affect minor salivary glands. We report a case of an intraductal papilloma arising in the unusual site of the submandibular gland. The tumour was completely excised and recurrence is not expected. A brief review of this histologically distinct lesion is presented.

Aged↗
PubMed1989

Effects of mammal submandibular gland glycoderivatives on human hemostatic system.

The aim of this study was to investigate the effects of glycoconjugate components extracted from the submandibular glands of four mammal species (rat, mouse, rabbit, hare) on human hemostatic system. The following analyses were performed: thromboelastography and hemocoagulation screening tests (Thrombin Time, Prothrombin Time, Partial Thromboplastin Time). Findings showed that all glycoconjugates induce modifications of fibrin clot formation time, modulus of elasticity and some hemocoagulation tests (TT, PTT). The anticoagulant effects were of inhibitory type.

Animals↗
PubMed2000

Sympathetic decentralization abolishes increased secretion of immunoglobulin A evoked by parasympathetic stimulation of rat submandibular glands.

Salivary secretion of immunoglobulin A (IgA) in response to electrical stimulation of the parasympathetic nerve supply was assessed bilaterally in the submandibular glands of anaesthetized rats 1 week following unilateral pre-ganglionic sympathectomy (decentralization). Nerve-mediated stimulation on the non-denervated side increased IgA secretion several fold above an unstimulated rate of secretion whereas sympathetic decentralization reduced the parasympathetically stimulated secretion of IgA without affecting the basal rate. Glandular levels of IgA were increased following decentralization compared to the control glands. Salivary levels of free secretory component (FSC), the cleaved polymeric immunoglobulin receptor (plgR), were increased by parasympathetic stimulation and reduced by sympathectomy, though not as much as IgA. The decreased secretion of FSC suggests a reduced production of plgR and may account in part, for reduced IgA secretion following long-term removal of sympathetic nerve impulses.

Animals↗
PubMed1983

Submandibular gland function following transoral sialolithectomy.

Conflicting views exist with regard to surgical treatment of a salivary stone in the knee of the submandibular duct, the options being excision of the submandibular gland and transoral removal of the stone. The key question in this dispute is whether or not a gland with a stone in this position has suffered irreparable functional damage. To determine the extent of glandular recovery after transoral sialolithectomy, a clinical and scintigraphic study has been carried out in a consecutive series of twenty-one patients. The results show a return to normal function in the vast majority of these patients, which indicates that transoral removal of the stone is the treatment of choice.

Adolescent↗
PubMed1996

Cytomegalovirus in the principal submandibular gland of the little brown bat, Myotis lucifugus.

In the course of a large scale study of salivary gland ultrastructure in chiropterans, enlarged cells infected with numerous virus particles were encountered in some acinar cells in the principal submandibular glands of two of 34 little brown bats (Myotis lucifugus). The characteristic morphology of the viruses, together with the cytomegaly that they induced, led to their identification as cytomegalovirus (CMV). In a reversal of the situation in other animal species, bat virus particles within cytoplasmic vacuoles lacked capsomeres, whereas the latter were prominent in particles free in the cytosol. The generally accepted schemes for CMV production cannot explain this seemingly aberrant morphology. This report extends to four the number of mammalian orders in which CMV has been documented by means of electron microscopy as occurring in salivary glands.

Animals↗
PubMed1989

[Regeneration of auto- and isografts of submandibular glands in the brain of laboratory mice. I. Morphologic evaluation of the healed graft using light microscopy].

The mouse granular convoluted tubules of submandibular gland of laboratory mouse are releasing a range of biologically active peptides (renin, neural growth factor and others) into both the saliva and blood circulation, the males producing it in a larger extent than females. Recently, several from respective peptides were identified to be endogenous ones and brain-related. The present work was aimed to utilise submandibular gland/SMG auto- and isotransplants regenerating in murine brain as a possibly local source of peptides. Experimentally, the newborn and juvenile matured white A breeded mice of both sexes were used. Glandular grafts were grafted into brain parenchyma or CSF spaces. Laboratory animals have then been perished during the first 6 weeks after transplantation, and the transplants so acquired evaluated as serial frontal sections embedded in paraffin and H.E. stained by light microscopy. Also cryocate sections were incubated in order to detect the presence of alkaline phosphatase (AP) and succinic dehydrogenase/(SDH). It was stated experimentally that both mentioned SMG grafts underwent the survival and development intracerebrally. Some first regressive changes were gradually replaced by glandular proliferation and lobular neomorphogenesis having been more pronounced in osotransplants. The proliferative period was characterized by cellular mitoses, multiplication of duct-like and terminal tubulous structures of newly formed glandular lobules. Partially, the isotransplants display the transformation of proliferation stage into that of cellular cytodifferentiation followed by gradual appearance of striated ducts, acini and even granular convoluted tubules on the 5th week after transplantation. Also the reoccurrence of enzyme activities in the transplant parenchyma after their initially total disappearance is testifying of both proliferation and cytodifferentiation developed gradually. During the first days of implantation, the revascularization of grafts occurs, those being high in AP endothelial activity of vessels newly formed. This is to conclude that higher proliferative intensity of isotransplants and their exclusive cytodifferentiation demonstrate that an undifferentiated murine SMG which can develop itself ontogenetically is more effective graft than a SMG differentiated fully. On the next stage, the development of glandular grafts will be studied with more delay after transplantations. Also the enzyme implementation of new parenchymatous components is to be elucidated. Further experimentation is planified as to influencing intracerebral SMG graft development with administration of hormones and isoproterenol to laboratory animals.

Animals↗
PubMed1982

Permeability of canine submandibular glands to blood-borne horseradish peroxidase during parasympathetic secretion.

Potential permeability pathways exist between central acinar cells in submandibular glands of dogs and they are large enough to allow the passage of molecules of horseradish peroxidase (HRP) under certain conditions. During gentle submandibular secretion caused by parasympathetic nerve stimulation detectable amounts of HRP passed into the saliva as long as the concentration of HRP in the blood reaching the gland was high enough. Morphological results indicated that the passage was mainly via leaky junctions between acinar cells. It is likely that this movement was by passive permeation because it required a high concentration gradient, was slow to occur and gradually reached a peak; the actual amounts to reach the saliva under physiological conditions were small and depended on the effective concentration of HRP reaching the gland. A satisfactory reproducible secretory regime has been developed, using short periods of stimulation alternating with rest pauses. Using these conditions gradually increasing amounts of HRP reached the saliva and first samples of saliva during each stimulation period contained more HRP than subsequent samples from the same stimulation period. It is possible that contraction of myoepithelial cells, at the outset of each stimulation period, contributed to this increased movement by acting on HRP-containing fluid in spaces between themselves and the acinar cells, thereby causing an increased filtration pressure to be exerted on underlying tight junctions.

Animals↗
PubMed1993

Fine structure and subepithelial capillaries of the main excretory duct epithelium of the submandibular gland of the Japanese monkey, Macaca fuscata.

The fine structure and subepithelial capillaries of the main excretory duct epithelium (MEDE) of the submandibular gland in the Japanese monkey, Macaca fuscata were investigated by light, scanning, and transmission electron microscopy. Capillaries found beneath the MEDE were all fenestrated ones. Some of them had elevated endothelium known as pored domes. The capillary density (number of capillaries per 200 microns length of MEDE) was 6.05 +/- 1.36, and the number of fenestrae per 10 microns length of available endothelium was 11.62 +/- 3.82. The MEDE was pseudostratified, and consisted of four cell types: Type I and II, which were columnar cells without basal infoldings, basal and goblet cells. The most characteristic feature was the presence of well-developed lateral interdigitations between Type I and II cells.

Animals↗
PubMed1998

[The inhibiting effect of glycyrrhizin on proliferation of the mice submandibular gland fibrosarcoma cell line in vitro].

OBJECTIVE: To study the inhibiting effect of glycyrrhizin on proliferation of the cell line of mice submandibular gland fibrosarcome in vitro. METHODS: Light and electronic microscope examination, proliferative state of cells and cycle phase analysis were used. RESULTS: Administration of glycyrrhizin above 600 mg/L could inhibit cell proliferation, and the effect was dose and time dependent. The morphological change of the cell line affected by glycyrrhizin could be seen with light and electronic microscope. Cell cycle phase analysis showed gLy-cyrrhizin gould stop cells shifting from G1 to S phase. CONCLUSION: The Lowest inhibition concentration of glycyrrhizin to the cell line was above 600 mg/L, The antineoplastic effect of glycyrrhizin was possibly accomplished by interfering metabolism of G1 phase cells or/and blocking initiation of DNA synthesis.

Animals↗
PubMed1987

Thyroidal modulation of androgenic expression in mice submandibular gland.

We investigated the influence of testes and thyroid gland on the maintenance of biochemical parameters and of [3H]R1881 binding sites of adult mice submandibular gland (SMG). Castration (Cx) performed at beginning of puberty prevented sex-dependent SMG development without interfering with maximal androgen binding capacity. Thyroidectomy (Tx) had strong effects on SMG, mainly by lowering the number of androgen binding sites. All alterations could be fully reverted after treatment with testosterone (5 mg/animal, single dose) or with thyroxine (T4, 250 micrograms/animal per day during 5 days). The effects of Cx on SMG could be reverted by therapy with testosterone, T4, or with both hormones (testosterone + T4) in a non-synergistic fashion. It is shown the importance of thyroidal activity on the physiological maintenance of androgen receptors in the murine SMG; the role played by thyroid gland seems to be essential for the full expression of the androgen-dependent SMG activity in adult mice.

Animals↗
PubMed1989

Light and electron microscopic immunolocalization of rat submandibular gland mucin glycoprotein and glutamine/glutamic acid-rich proteins.

We studied the subcellular localization of two major secretory products of adult rat submandibular gland (RSMG), blood group A-reactive mucin glycoprotein and glutamine/glutamic acid-rich protein (GRP), by light and electron microscopic immunocytochemistry. The structure of the major neutral oligosaccharide of the mucin was shown to be: GalNAc alpha 1,3(Fuc alpha 1,2)Gal beta 1,3GalNAc. A mouse monoclonal antibody (1F9) with specificity for blood group A determinants was prepared against the mucin. The antibody recognized a single band of approximately 114 KD on Western blots of RSMG extract. A previously characterized monoclonal antibody (59) against GRP (Mirels et al.: J Biol Chem 262: 7289, 1987) reacted with a doublet of 45-50 KD on Western blots of extraparotid saliva. Immunofluorescence and immunoperoxidase staining of cryostat sections of RSMG with anti-mucin antibodies and anti-GRP antibodies revealed reactivity in acinar cells of the gland. No specific labeling was seen in duct cells of RSMG or in mucous acinar cells of the adjacent sublingual gland. Post-embedding immunogold labeling of thin sections of glutaraldehyde-fixed RSMG with anti-mucin showed strong labeling of the Golgi apparatus and secretory granules of acinar cells. Gold particles were seen mainly over electron-lucent areas of the granules. No labeling occurred over the endoplasmic reticulum. The labeling pattern with the anti-GRP antibodies was similar, except that both electron-dense and -lucent areas of the granules were labeled, and the endoplasmic reticulum was reactive. Double labeling with two different sizes of gold particles showed that both mucin and GRP co-localized in the same granules. Pre-absorption of the antibodies with their respective antigens eliminated immunolabeling of the acinar cells. These antibodies will be useful in studies of cell differentiation in RSMG and of synthesis, processing, and packaging of RSMG secretory products.

Animals↗
PubMed2005

Submandibular gland myoepithelioma.

Benign myoepithelioma is a very rare form of salivary gland tumor, composed entirely of myoepithelial cells. It accounts for approximately 1% of all salivary gland tumors and is most frequently located in the parotid gland and in the minor salivary glands of the hard palate. We describe herein the ninth reported case of myoepithelioma of the submandibular gland. Benign myoepithelioma must be differentiated from several benign and malignant epithelial and mesenchymal tumors. Immunohistochemical staining can help differentiate between these conditions, but histopathology remains the gold standard for diagnosing this neoplastic process.

Diagnosis, Differential↗