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PubMed1989

Intranuclear androgen-receptor complex binding sites of mouse submandibular gland.

Nuclear androgen-receptor binding sites in chromatin regions of mouse submandibular gland were studied. Cytosol androgen receptors prelabeled with [3H]androgen interacted with the crude nuclei in mouse submandibular gland and activation of the receptor was a prerequisite for this interaction. After in vivo administration of [3H]androgen to female mice, radioactivities were found in the nuclei purified from submandibular gland tissues and the [3H]androgen-labeled purified nuclei were further digested with micrococcal nuclease. The androgen receptor was found in solubilized, active chromatin fractions which contained mono- and dinucleosomes. By an in vitro exchange assay, endogenous androgen-receptor complexes associated with chromatin binding sites in intact males were found in the solubilized fraction after micrococcal nuclear digestion, whereas such complexes were not found in females. These results suggest that the androgen receptors translocated to the nuclei and became associated with chromatin and that this association occurred in transcriptionally active chromatin regions that were preferentially sensitive to micrococcal nuclease.

Animals↗

PubMed1984

Adrenergic innervation and noradrenaline content of the rat submandibular gland during the experimental Trypanosoma cruzi infection.

Rat submandibular glands were tested histochemically for the presence of formaldehyde induced fluorescence 18, 32, 45, 60 and 100 days after inoculation with Y strain of Trypanosoma cruzi. At days 32, 60 and 100 the glandular noradrenaline content was also assayed fluorometrically. The fluorescent varicose nerve fibres of all the studied glands were severely reduced at days 18 and 32 of infection. At day 45 reduction was not so pronounced and by day 60 the density of adrenergic terminals was similar to that of the control glands. At day 32 of infection the glandular contents of noradrenaline expressed as ug/gland or ug/g of fresh tissue dropped to 50% of the control values. At day 64, the noradrenaline contents per gland were still different from the controls. However, at day 100 no difference was observed between the values for control and infected animals. These results indicate that the sympathetic innervation of the rat submandibular gland is affected during the acute phase of Chagas' disease but returns to normality during the chronic phase.

Adrenergic Fibers↗

PubMed1995

Cell proliferation and apoptosis in isoprenaline-induced sialosis in the rat submandibular glands.

Regression of submandibular acinar cell hyperplasia after withdrawal of the mitogenic stimulus induced by isoprenaline was studied in male Wistar rats. Intraperitoneal administration of 39 mg of isoprenaline, in divided doses over a 9-day period, resulted in a marked increase in gland size and weight. The proportional volume occupied by acinar cells increased. These changes were associated with acinar cell proliferation and an increase in the mitotic index and cytoplasmic: nuclear ratio. Thus, both hyperplasia and hypertrophy clearly contributed to gland enlargement. Granular and other duct cells appeared to be unaffected by isoprenaline administration. Following cessation of the drug, the mitotic index reached a peak value by the second day but thereafter declined rapidly. This change was matched by a marked rise in the apoptotic cell index, the latter reaching a maximum by the fourth day. By the end of the second week, glands had returned to normal with respect to size and weight and neither hyperplasia nor apoptosis could be detected histologically. These results confirm that apoptosis is involved in the regulation of submandibular salivary gland size.

Animals↗

PubMed2002

Ultrastructure of the submandibular gland of the rare white-winged vampire bat, Diaemus youngi.

The submandibular gland of the white-winged vampire bat, Diaemus youngi, was examined by electron microscopy. Unlike typical submandibular glands, those in Diaemus have only one type of secretory cell in their endpieces, namely, serous cells. These serous cells are conventional in structure, with an extensive rough endoplasmic reticulum, scattered dictyosomes, and numerous secretory granules. The endpiece lumina, as well as intercellular canaliculi, are fitted with numerous microvilli, which also are present on the otherwise unremarkable intercalated duct cells. Striated ducts are of conventional morphology, but have a brush border-like array of microvilli on their luminal surface. These cells resemble those in the submandibular gland of the common vampire bat, Desmodus rotundus. The presence of an abundance of microvilli in the salivary glands in the two vampire bat species (and their absence from chiropteran species that consume other types of diets) is a strong indication that these structures play a significant role in dealing with the problems posed by a sanguivorous diet.

Animals↗

PubMed2004

Cell death and cell proliferation in the regeneration of atrophied rat submandibular glands after duct ligation.

BACKGROUND: The present study aimed to clarify the proliferation and apoptosis of parenchymal cells during regeneration of rat submandibular glands following atrophy. METHODS: Atrophy of the right submandibular gland of rats was induced by excretory duct ligation at the hilum with metal clips, which were removed 1 week (day 0) after ligation. The right submandibular glands were collected from 0 to 14 days after removal of the clips and investigated using immunohistochemistry for proliferating cell nuclear antigen (PCNA) as a marker of proliferating cells, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-digoxigenin nick end labeling (TUNEL) as a marker of apoptotic cells, and transmission electron microscopy (TEM). RESULTS: After 1 week of ligation, there were many remaining ducts and a few acini in the atrophic glands. At day 3 after discontinuing the ligation, newly formed acini appeared and thereafter increased in number and maturity. Many residual and newly formed acinar cells showed positive reaction to PCNA especially at days 4 and 5. The PCNA-positive duct cells decreased in number with the regeneration. A few TUNEL-positive acinar and duct cells were identified during regeneration. Mitosis and apoptosis of parenchymal cells were also identified by TEM. CONCLUSIONS: During regeneration of the submandibular gland after atrophy, both residual and newly formed acinar cells proliferate actively. There is also apoptosis of parenchymal cells; however, the significance of apoptosis is low.

Animals↗

PubMed2002

Taurine in submandibular gland of the rat: effect of muscarinic drugs.

Taurine exerts a number of actions in mammalian cells, including regulation of ion transport and osmoregulation. The production and secretion of saliva involve transepithelial ion transport, thereby making the plasma-like primary saliva hypotonic before secretion. Therefore, it is plausible to suggest modulation of salivary taurine by muscarinic agents that affect salivary gland function. One of the objectives of this study was to determine tissue content and localization of taurine in the submandibular gland of the rat. Further, we determined whether treatment with muscarinic drugs that either increase (e.g., pilocarpine) or decrease (e.g., propantheline) saliva secretion affects the submandibular gland taurine content. The results indicate that the submandibular gland contains an appreciable amount of taurine (8.9 +/- 0.3 micromoles/g wet wt). Further, acute treatment of the rats with either of the muscarinic drugs did not significantly affect tissue taurine content compared to the control group. By contrast, chronic treatment with propantheline, but not pilocarpine, reduced the tissue content of taurine compared to the control rats (p<0.05). Utilizing light microscopic immunohistochemical techniques, intense immunoreactivity was found primarily in the striated ducts of the submandibular gland. Neither pilocarpine nor propantheline treatment led to differential distribution of immunoreactivity in this tissue. In conclusion, the submandibular gland contains an appreciable amount of taurine, primarily in the striated ducts, that can be decreased by chronic muscarinic receptor blockade.

Animals↗

PubMed1990

Morphological study of granular convoluted tubules in the submandibular gland of the mouse during the growth of a sarcoma-180 subcutaneous tumor.

Histological and cytological changes in the submandibular glands of adult male mice arising during the growth of sarcoma-180 subcutaneous tumors were studied. The submandibular glands of the mice were examined by morphometric analysis at 1, 3, 6, 10, 20, 30 and 64 days after inoculation of the tumor cells. There was a slow increase in the relative cross-sectional area of the granular convoluted tubule (GCT) in the section of the submandibular gland of the animals as the tumors grew. The increased proportional area of the GCT was significantly different from that of the control's from day 30. However, the mean weight of the glands was not increased. The proportional area of the granular cluster in the cells of the GCT of tumor cells in inoculated animals decreased about 5% on the first day and then quickly increased by 16% on the third day in comparison with those of the controls, eventually reaching a maximum of 74% (control, 54%) by day 30. In addition, the average number of granules per GCT cell decreased in the first three days, then increased to normal levels from day 6, going above the normal level from day 20 of the tumor growth. These changes in the glands of tumor-bearing animals disappeared within 20 days after removal of the tumor. These results indicate that the growth of the sarcoma-180 subcutaneous tumor caused morphological changes in the GCT and GCT cells, suggesting an alternation in the requirements of the secretions contained in the granules, such as the epidermal growth factor, during the growth of the tumor.

Animals↗

PubMed2005

The effect of local injection of botulinum toxin A on the immunoreactivity of nNOS in the rat submandibular gland: an immunohistochemical study.

PURPOSE: In our study, we intend to investigate the influence of local injections of botulinum toxin A on the activity of neuronal nitric oxide synthase (nNOS) in submandibular glands of adult rats. Since interest has been focused on the role of nitric oxide (NO) as a possible neuromodulator of secretory regulation processes in the upper aerodigestive tract, it was the aim of the present study to show that the toxin also interferes with the metabolic actions of NO on investigated rat submandibular glands. It is of great clinical interest whether the NO pathway is able to influence salivary gland secretion. Increasing of knowledge in this field maybe helpful to treat sialorrhoea, especially in juvenile otolaryngologic and neurologic patients. MATERIALS AND METHODS: We performed immunohistochemical reaction of neuronal nitric oxide synthase (nNOS) in the submandibular gland of female adult Wistar rats, both in native (untreated) glands and after intraglandular injection of botulinum toxin A under general anesthesia. The immunoreactivity of nNOS was investigated on different times after injection. RESULTS: Other than in the untreated glands, there was a significant decrease of nNOS in the treated organs, which became stronger with extended toxin exposure time. The present study shows explicit data on the effect of botulinum toxin A injection on a higher number of examined submandibular glands and is able to analyze a time course of the effect duration. CONCLUSION: In our study, it was shown that botulinum toxin A had an influence on the immunoreactivity of neuronal nitric oxide synthase (nNOS) in submandibular glands. Therefore, the participation of nitric oxide (NO) in the regulation of secretion from these organs seems to be evident. It might be assumed that the influence of botulinum toxin A on nNOS in the submandibular gland of the rat is able to explain the sometimes longer duration of toxin effect at the neuroglandular junction than at the motor endplate.

Animals↗

PubMed1989

Mechanism of action of calcitonin on secretion in rat submandibular gland.

Calcitonin (CT) was found to reduce the initial flow of pilocarpine-stimulated saliva from the submandibular glands in the rat. Although there was a concomitant increase of the concentration of calcium and protein in the saliva, the calcium/protein ratio was not significantly affected. CT also caused a significant increase of the potassium concentration in submandibular saliva. Both in vivo and in vitro, CT inhibited the production of cyclic AMP (cAMP) both in the absence and in the presence of forskolin. This decrease in intracellular cAMP levels could result in an inhibition of mucus secretion, which would explain the previously observed calcitonin-induced intracellular accumulation of mucus in the submandibular gland acinar cells. CT did not affect the cytoplasmic free Ca2+ concentration (as measured with fura 2) in isolated submandibular acini either in the absence or in the presence of cholinergic or adrenergic agonists. These results indicate that the inhibition of fluid secretion in the submandibular gland by calcitonin must be located distal to changes in [Ca2+]i. It can be concluded that CT affects both mucus and fluid secretion in the submandibular gland, but that only the inhibition of mucus secretion can as yet be explained by an effect at the level of the second messenger.

Adenylyl Cyclases↗

PubMed1989

[A quantitative morphologic study on age changes in the histologic structures of the human submandibular gland].

One hundred and nine samples of human submandibular glands were studied by quantitative morphometric methods. All of the samples in this study were removed during operations. According to the age distribution of patients, 109 samples were divided into the young (less than 25 years), the adult (25-44 years), the elderly (45-59 years) and the aged (greater than or equal to 60 years). The results had shown that the amount of the proportional volume of parenchymal cells, comparing with that of the young, decreased 23 percent in the aged, the proportional volume of acini tissue decreased 31.2%, the proportional volume of the fat and fibrous tissues increased 2.0 and 0.8 times, respectively. The relationship between aging and changes of the proportional volume of parenchymal cells, duct cells, fat and fibrous tissues in human submandibular glands were discussed.

Adolescent↗

PubMed1986

Effects of VIP, PHM and substance P on blood vessels and secretory elements of the human submandibular gland.

The effects of the neuropeptides VIP, PHM and substance P (SP) on vascular smooth muscle tone, K+ secretion from exocrine elements and tissue content of cyclic AMP (cAMP) in the human submandibular gland were studied in vitro. All three peptides caused relaxation of noradrenaline contracted human submandibular arteries at nM concentrations. SP was slightly more active than VIP and PHM which had a similar potency as vasodilators. Only carbachol but not VIP, PHM or SP stimulated K+ secretion from exocrine elements of the human submandibular gland. Principally similar in vitro effects on K+ secretion were obtained on the cat submandibular gland, but in the rat not only carbachol but also SP stimulated K+ secretion. VIP and PHM increased cAMP production of exocrine elements in the human submandibular gland in nM concentrations. VIP was about 5-fold more potent than PHM with regards to cAMP production. In conclusion, VIP, PHM and SP relaxed human submandibular arteries in vitro. Both VIP and PHM stimulated cAMP production in glandular tissue but none of the three peptides induced K+ secretion from human submandibular gland tissue. This suggests that, in contrast to the situation in the rat, SP does not cause watery salivation in man, while VIP and PHM may modulate protein e.g. amylase content of the saliva.

Adult↗

PubMed1992

Molecular cloning and characterization of rKlk10, a cDNA encoding T-kininogenase from rat submandibular gland and kidney.

We have cloned and determined the nucleotide sequence of a novel kallikrein-like mRNA, designated rKlk10*, from rat submandibular gland and kidney with the aid of the polymerase chain reaction (PCR). This cDNA contains 737 base pairs comprising the sequence encoding a mature protein of 235 amino acid residues, partial zymogen peptide, and 3' noncoding sequence. Sequence comparisons showed that rKlk10 mRNA shares 87 and 88% sequence identity with rat tissue kallikrein at nucleic acid and amino acid levels, respectively. It encodes a 26,428-Da acidic protein whose derived amino acid sequence matches completely with the partial amino acid sequence of a kallikrein-like enzyme designated as T-kininogenase, K10 protein, or antigen-gamma purified from rat submandibular gland [Xiong et al. (1990) J. Biol. Chem. 265, 2822-2827; Gutman et al. (1991) Eur. J. Biochem. 784, 1-5; Berg et al. (1991) Biochem. J. 280, 19-25]. The protein encoded by rKlk10 retains the key amino acid residues determining kallikrein cleavage specificity. Northern blot analysis with an rKlk10-specific oligonucleotide probe showed that its mRNA level in the submandibular gland is decreased dramatically by administration of the beta agonist isoproterenol. Tissue-specific expression of rKlk10 was analyzed by Northern blotting and Southern blotting of PCR-amplified cDNA, which showed that rKlk10 is expressed at high levels in the submandibular gland and low levels in the kidney but not in seven other tissues including prostate, liver, heart, adrenal gland, testes, pituitary, and pancreas. rKlk10 cDNAs cloned from the kidney and submandibular gland show sequence identity.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗

PubMed2001

Stereological study on the submandibular gland in hypothyroid rats.

Most research done on hypothyroidism has focused on physiological and biochemical aspects of the tissues, whilst there has been little work on tissue morphology especially on salivary glands. The present study has used hypothyroid Wistar rats as a model for investigating the effects of hypothyroidism on submandibular gland structure. Two groups of Wistar rats were studied. One was made hypothyroid with methimazole and the second was an untreated euthyroid group (control). They were euthanised after 10 weeks. Submandibular glands were removed and studied. Systematic random transverse sections were obtained from submandibular glands and subjected to morphometric analysis. Volume density and absolute volume of granular, striated and excretory ducts and interlobular connective tissue were estimated by point counting. Volume-weighted mean particle volume of serous and mucous acini was also determined by unbiased stereology. Absolute volume of granular ducts in the submandibular gland of hypothyroid rats was reduced by approximately 50% (p<0.009) whilst that of the striated and excretory ducts and interlobular connective tissue was unaffected. Volume of serous acini was also significantly (p<0.03) lower in hypothyroid rats. These changes suggest that hypothyroidism has an effect on submandibular gland structure, and that this effect occurs mostly in two major exocrine compartments (granular duct and serous acinus) of the gland.

Animals↗

PubMed2006

Minimally invasive endoscopic resection of the submandibular gland: a new approach.

BACKGROUND: This study evaluates the benefits of a new approach, endoscopic resection of the submandibular gland through the hyoid midline level skin incision by use of an ultrasonically activated scalpel. METHODS: Twelve operations for patients presenting with submandibular gland disease were performed via minimally invasive endoscopic resection by a single surgeon (M-KC). RESULTS: All 12 submandibular gland resections were successfully performed endoscopically, and no conversions to conventional open resection were necessary. Of the 12 patients who underwent excision, 3, 6, and 3 had mixed tumor, sialoadenitis with sialolithiasis, and chronic sialoadenitis, respectively. The procedures lasted 50 to 125 minutes (median duration, 70 minutes). No complications associated with the operation occurred, and the scar was almost invisible because of its concealed location with the neck in the natural position. CONCLUSIONS: Endoscopic resection of the submandibular gland is a feasible method for treatment of benign lesions. The main advantages of this procedure are that the small operative scar is concealed in the submental skin crease, resulting in improved cosmetic results and minimization of the possibility of facial nerve injury.

Adult↗

PubMed1983

Malignant lymphoepithelial lesion of the submandibular gland.

A case of malignant lymphoepithelial lesion affecting the submandibular gland of a 48-year-old Japanese man is described. Histologically, a well-encapsulated tumorous mass replacing almost the whole gland was composed of lymphoid and epithelial elements, and the latter revealed severe anaplasia with frequent mitotic figures indicating malignant changes. There is no sign of recurrence up to now, 32 months after the operation. Ultrastructural studies on a formalin-fixed sampling from the gland demonstrated the squamous nature of the epithelial component. This is the first case of malignant lymphoepithelial lesion of the submandibular gland detected in Japan.

Epithelium↗

PubMed1996

Kininogenase secretion from dispersed cells of mouse submandibular gland.

Kininogenase activity in both male and female submandibular gland (SMG) began to increase at 4 weeks of age and reached a peak at 6 weeks of age, and then showed a stable level of activity until 10 weeks of age. The release of kininogenase was stimulated via activation of alpha-adrenoceptors alone in male SMG and via both alpha-adrenoceptors and muscarinic cholinergic receptors in female SMG. This indicates that there is a marked difference between male and female SMG in response to methacholine, e.g., the less granular convoluted tubule (GCT) there is in the SMG, the higher is the percentage release of kininogenase evoked by methacholine. It is suggested from these data that the site of localization of kininogenase in the male and female mouse submandibular gland is different; probably predominantly in the GCT in the male and in the striated duct in the female.

Adrenergic alpha-Agonists↗

PubMed1990

[Release mechanisms of inactive renin from rat renal cortical slices: role of the submandibular gland].

In our previous studies, we showed an in vivo stimulating effect of the extract of the rat submandibular gland on plasma inactive renin release. In this study, we evaluated the effects of the rat submandibular gland extract and of some plasma active renin stimulants on inactive renin release from rat renal cortical slices. Adult male Wistar rats (250-350g) were kept on a regular diet (Na 260mg/100g) and nephrectomized under pentobarbital anesthesia (50mg/kg, i.p.). Five thin renal cortical slices were obtained from each kidney by using a razor blade. These renal cortical slices were incubated in Earle's buffer (pH7.4, Difco) at 37 degrees C for 30 min (preincubation), then transferred into 10ml fresh Earle's buffer with or without some agents and incubated at 37 degrees C for 1 hour (experimental incubation). For each experiment, 6 groups of 5 renal cortical slices were employed. The agents used in this study were as follows: isoproterenol (10(-5)M), furosemide (50 micrograms/ml), prostaglandin E1 (10(-5)M), prostaglandin I2 (10(-5)M) and the rat submandibular gland extract (100 microliters) which was obtained after homogenation with 10 x (w/v) 0.01M pyrophosphate buffer (pH6.5) including 0.1M NaCl. One ml of samples of this Earle's buffer were withdrawn every 20 min. Active renin in the samples was assayed by the commercial RIA-kit (Dainabot), and total renin was assayed after trypsin (Worthington) treatment (30 micrograms/300 microliters sample) at 4 degrees C for 10 min. Inactive renin was determined as the difference between total renin and active renin. Active and inactive renins increased linearly in the buffer without any agents (control) during the observation period (60 min). Isoproterenol (10(-5)M) stimulated the release of active renin significantly (p less than 0.01 vs. control) but did not affect the release of inactive renin. Furosemide (50 micrograms/ml) stimulated the release of active and inactive renins significantly at 20 and 40 min (p less than 0.05 vs. control) but did not affect the release of either renin at 60 min. Both prostaglandins E1 and I2 (10(-5)M) stimulated the release of active renin significantly (p less than 0.01 vs. control) but inhibited, on the other hand, the release of inactive renin significantly (p less than 0.01 vs. control). The rat submandibular gland extract (100 microliters) did not affect the release of active renin but stimulated the release of inactive renin significantly (p less than 0.05 vs. control).(ABSTRACT TRUNCATED AT 400 WORDS)

Alprostadil↗

PubMed1983

Characterization of the submandibular gland microsomal calcium transport system.

Calcium accumulation by submandibular gland microsomes (first described by Selinger and Naim, ((1970) Biochim. Biophys. Acta 323, 337-341) has been further characterized. Accumulation was concentration dependent, had a Km of 25 microM added calcium and a Vmax of 12 nM calcium/mg protein per min. No accumulation was observed in the presence of either the calcium ionophore A23187, or the detergent Triton X-100 (0.05). The divalent cations Sr2+ and Mn2+ inhibited accumulation competitively with Ki values of 67 microM and 200 microM, respectively. The effect of various enzyme inhibitors were tested on the microsomal calcium transport system and it was found that chlorpromazine, trifluoperazine, and DIDS all inhibited. The mitochondrial transport inhibitors ruthenium red and CCCP had no effect on transport. Experiments directed at clarifying the cellular location of the system are described. It was found that the membrane vesicles responsible for transport show different purification properties than the membrane vesicles which contain the standard enzyme markers for total and rough endoplasmic reticulum, Golgi apparatus, plasma membrane, and lysosomes. These conclusions are based upon experiments using these properties for membrane purification, density, size, and electrophoretic mobility. Three possible explanations of the results are given and it is organelles. The significance of the results in: (1) understanding the biochemical properties of the submandibular gland microsomal calcium transport system, (2) clarifying the cellular location of the system, and (3) clarifying the function of the system in salivary secretion are discussed.

Animals↗