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PubMed2002

Swelling-induced Ca2+ release from intracellular calcium stores in rat submandibular gland acinar cells.

The effects of osmotically-induced cell swelling on cytoplasmic free Ca2+ concentration ([Ca2+]i) were studied in acinar cells from rat submandibular gland using microspectrofluorimetry. Video-imaging techniques were also used to measure cell volume. Hypotonic stress (78% control tonicity) caused rapid cell swelling reaching a maximum relative volume of 1.78 +/- 0.05 (n = 5) compared to control. This swelling was followed by regulatory volume decrease, since relative cell volume decreased significantly to 1.61 +/- 0.08 (n = 5) after 10 min exposure to hypotonic medium. Osmotically induced cell swelling evoked by medium of either 78% or 66% tonicity caused a biphasic increase of [Ca2+]i. The rapid phase of this increase in [Ca2+]i was due to release of Ca2 + from intracellular stores, since it was also observed in cells bathed in Ca2+-free solution. The peak increase of [Ca2+]i induced by cell swelling was 3.40 +/- 0.49 (Fura-2 F340/F380 fluorescence ratio, n = 11) and 3.17 +/- 0.43 (n = 17) in the presence and the absence of extracellular Ca2+, respectively, corresponding to an absolute [Ca2+]i of around 1 microm. We found that around two-thirds of cells tested still showed some swelling-induced Ca2+ release (SICR) even after maximal concentrations (10(-5) M - 10(-4) M) of carbachol had been applied to empty agonist-sensitive intracellular Ca2+ stores. This result was confirmed and extended using thapsigargin to deplete intracellular Ca2+ pools. Hypotonic shock still raised [Ca2+]i in cells pretreated with thapsigargin, confirming that at least some SICR occurred from agonist-insensitive stores. Furthermore, SICR was largely inhibited by pretreatment of cells with carbonyl cyanide m-cholorophenyl hydrazone (CCCP) or ruthenium red, inhibitors of mitochondrial Ca2+ uptake. Our results suggest that the increase in [Ca2+]i, which underlies regulatory volume decrease in submandibular acinar cells, results from release of Ca2+ from both agonist-sensitive and mitochondrial Ca2+ stores.

Animals↗
PubMed1979

Effects of in vivo single and multiple isoproterenol injections on subsequently explanted submandibular glands.

The present study reports on the effects of in vivo single and multiple isoproterenol (ISP) injections on the proliferative response of subsequently explanted mouse submandibular glands. Explants were cultured in Waymouth's medium supplemented with 10% fetal bovine serum, and were harvested after 1,2,4 or 6 days in culture. 24 h prior to harvest, the explants were fed medium containing [3H] thymidine. At the time of harvest, the explants were fixed in situ and processed for light-microscopic radioautography. The number of labeled nuclei and the total number of nuclei were counted, and labeling indices were calculated for the control and ISP-treated explant sectons. The results show that the ISP-treated explants had more cells actively synthesizing DNA than did the corresponding control explants after 1 day in culture. In addition, after 2 days in culture, the explants from mice that had received a single dose of ISP incorporated more [3H] thymidine than the corresponding control explants. These results suggest that the in vivo ISP stimulus initiated a hyperplastic response in vivo that was completed during the 1st or 2nd day in vitro.

Animals↗
PubMed1991

Ultrastructure of the submandibular gland in the African multimammate rodent, Praomys natalensis.

Praomys natalensis, an African rodent that is phenotypically and cytogenetically intermediate to rats and mice, possesses a submandibular gland that is histologically similar to that in both of these near relatives, but is ultrastructurally unique. Acinar cells, which are seromucous in nature, contain secretory granules that often contain a perfect "bull's eye" inclusion (or some variant of this configuration) suspended in a dense matrix. The Golgi apparatus in these cells has an unusual structure, with the Golgi saccules often being doubled over, so that the outermost saccule also is the innermost. This peculiar architecture apparently arises fairly late in the secretory process, i.e., a Golgi apparatus of conventional structure gives rise to a nascent granule (condensing vacuole), then its saccules secondarily fold over. Intercalated ducts are preceded by a ring of specialized cells that have a number of serous-type granules, the duct cells themselves being devoid of such granules. Granular convoluted tubules (GCT) contain large dense granules that appear to be spontaneously involved in chain exocytosis. These GCT granules probably are the repositories of nerve growth factor, which is particularly abundant in Praomys. Striated ducts for the most part are typical in appearance, but they and, to a lesser extent, GCTs contain prominent, membrane-bound crystalloids with a periodicity of about 15 nm.

Animals↗
PubMed1995

Substrate specificity of microsomal 1-acyl-sn-glycero-3-phosphoinositol acyltransferase in rat submandibular gland for polyunsaturated long-chain acyl-CoAs.

Microsomal 1-acyl-sn-glycero-3-phosphoinositol (1-acyl-GPI) acyltransferase in the rat submandibular gland showed the highest specific activities for eicosanoid-related polyunsaturated acyl-CoAs, such as arachidonoyl-, bishomo-gamma-linolenoyl- and 5,8,11,14,17-eicosapentaenoyl-CoAs, with low Km values. High activities were also obtained with acyl-CoAs having long (more than 14 carbon atoms) and n - 6 unsaturated (more than 3 double bonds) acyl chains. This enzyme also utilized acyl-CoAs having trans-unsaturated or branched chains, but not short-chains, as substrates, although the activity levels for trans-unsaturated acyl-CoAs were lower than those for cis-unsaturated acyl-CoAs. Chronic administration of isoproterenol induced decreases of this enzyme activity and the content of arachidonic, bishomo-gamma-linolenic and 5,8,11,14,17-eicosapentaenoic acids at the sn-2 position of phosphatidylinositol. These results suggest that enrichment of arachidonic acid in the sn-2 position of phosphatidylinositol is established by the high specificity and affinity of 1-acyl-GPI acyltransferase for arachidonoyl-CoA. On the other hand, the low level of bishomo-gamma-linolenic and 5,8,11,14,17-eicosapentaenoic acids in the sn-2 position of phosphatidylinositol may be explained by their limited availability.

Acyl Coenzyme A↗
PubMed1995

Modification of the beta-adrenergic sensitivity of rat submandibular gland by constant light.

Rats exposed for 2, 5, 10, 20, 35 and 50 days to constant light (CL) showed beta-adrenergic desensitization of the submandibular gland as indicated by dose-response curves to 1.5, 2.0, 3.0, 10.0, 30.0 and 50.0 micrograms/kg isoproterenol. The phenomenon, evident with 1.5, 2.0 and 50.0 micrograms/kg after 2 days at CL, was more intense as time of exposure increased to reach values of about 50% those for controls after 20 days. After 35 and 50 days, desensitization showed some reversion, but the secretory responses were still lower than for controls maintained under a typical photoperiod. In the groups of rats exposed to CL for 20 days, gland wet and dry weights were higher (10%) than those of controls. These changes probably indicate an adaptation of the sympathetic control of salivary secretion induced by environmental illumination.

Animals↗
PubMed1986

Thyroxine increases neonatal mouse submandibular gland mRNA-directed synthesis of epidermal growth factor.

Thyroid hormones are known to modulate the concentrations of epidermal growth factor (EGF) in the mouse submandibular gland (SMG); this action is presumably mediated by the nuclear triiodothyronine receptor. To test the hypothesis that thyroid hormones act to increase SMG EGF concentrations by increasing the number of poly(A)+ -specific mRNA, poly(A)+ RNA was isolated from SMGs of neonatal mice which had been treated daily from birth through to 21 days of age with thyroxine (T4,0.4 microgram/g body weight). Poly(A)+ RNA also was extracted from SMGs of intact 21-day-old mice which had received vehicle alone. No significant differences in total nucleic acid, total RNA, or poly(A)+ RNA yields were noted between the two groups of animals. The isolated poly(A)+ RNAs from T4-treated and control mice were translated in an in vitro wheat germ system. Although no significant differences in efficiency of [35S]cysteine incorporation into trichloracetic acid precipitable material were noted between the two poly(A)+ RNA preparations, a significantly greater proportion of radioactivity was immunoprecipitable by anti-EGF antiserum in the translation medium derived from T4-treated mice (17.2 +/- 0.9%, mean +/- SEM) than in that of control mice (7.3 +/- 0.5%, P less than 0.001). Polyacrylamide gel electrophoresis of the immunoprecipitates (IMMP) revealed the presence of three radioactive bands with apparent relative masses (MrS) of 12,000, 9000, and 6000. The latter species comigrated with purified EGF, [125I]EGF, and an IMMP of a SMG extract. The translation product IMMPs following polyacrylamide gel electrophoresis were iodinated and digested with alpha-chymotrypsin.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1998

Time-dependent changes of levels of endogenous epidermal growth factor in submandibular glands, in kidneys, and in urine in rats during systemic treatment with EGF.

BACKGROUND: Exogenous EGF influences the levels of endogenous EGF differently in the submandibular glands (SMG) and the kidneys. The aim of the present study was to examine the time-dependent changes in levels of endogenous EGF during 1-4 weeks of EGF treatment. METHODS: Female rats were allocated into five groups receiving EGF subcutaneously (150 microg/kg/day) for 0 (controls), 1, 2, 3 and 4 weeks prior to sacrifice at an age of 12 weeks. At the end of the study period, 24-h urine samples were collected. RESULTS: The weight of the SMG increased during EGF treatment (303+/-33 (controls), 359+/-37 (1 week EGF, P < 0.01), 390+/-30 (4 weeks EGF, P < 0.001) (mg mean+/-S.D.)). The EGF content of the SMG was unchanged after 1 week but threefold decreased after 4 weeks of treatment, respectively. The expression of EGF mRNA was decreased after 1 and 4 weeks as assessed with in situ hybridization. The weight of the kidneys was unchanged after 1 week and increased after 4 weeks of treatment (828+/-105 mg (controls) vs. 935+/-44 mg (4 weeks EGF, P < 0.005)). The renal content and the urinary excretion of EGF were significantly increased by 20-30% only in the group treated for 4 weeks. CONCLUSION: EGF treatment induces a time-dependent decrease in the EGF content in the SMG most likely by reducing the biosynthesis of endogenous EGF. In contrast, the EGF content in kidneys and in urine was unchanged after 1 week and increased after prolonged treatment.

Animals↗
PubMed1995

Regulation by purinergic agonists of zinc uptake by rat submandibular glands.

The zinc uptake in rat submandibular cells was measured using fura2 as a fluorescent probe. Basal zinc uptake was observed in a 100 microM - 1 mM concentration range. Carbachol and isoproterenol had no effect but ATP4- dose-dependently increased the basal zinc uptake (half-maximal concentration: 250 microM). The purinergic agonist shifted the concentration curve for zinc to the left by one order of magnitude. The response to ATP was not reproduced by adenosine or ADP and was blocked by Coomassie blue. Calcium, nickel or lanthanum were inhibitors of zinc uptake, while the substitution of extracellular sodium by potassium or lithium increased the basal zinc uptake. We conclude that in submandibular cells zinc can permeate through the non-specific cation channel coupled to ATP-sensitive purinergic receptors.

Adenosine↗
PubMed1975

[Identification of a methionine residue as the reactive site for chymotrypsin in the double-headed proteinase inhibitor from the canine submandibular gland (author's transl)].

The canine submandibular inhibitor is double-headed with two independent reactive sites. Whereas the trypsin-reactive center (-Ala-Cys-Pro-Arg26-Leu-His-) is located in domain I, the chymotrypsin-reactive site (-Met-Cys-Thr-Met78-Asp-Tyr-) is located in domain II. The presence of a methionine residue in this inhibition center is supported by the findings that nitration with tetranitromethane abolishes neither trypsin nor chymotrypsin inhibition, whereas after alkylation of the methione residues, only trypsin inhibition is retained. Remarkably, another inhibitor from microbial sources [10] which also contains a methionine residue in the presumed reactive site also inhibits subtilisin but not chymotrypsin (or trypsin).

Amino Acid Sequence↗
PubMed1979

Microcinematographic demonstration of synchronous and asynchronous myoepithelial contractions in mouse submandibular gland rudiments in organotypic culture.

Time-lapse phase-contrast cinematography revealed contractile activity within mouse submandibular salivary gland rudiments in organotypic culture. Three types of contraction were distinguishable. In type I (voiding contractions), all portions of the gland contracted synchronously, and the active state ranged from 30 min to 2 hr. In type II (priming contractions), all portions of the gland contracted synchronously, but the active state was shorter, ranging from 4 to 10 min. In type III (churning contractions), isolated foci in lobules or secretory units throughout the gland contracted asynchronously and had very short active states of about 1 min. By electron microscopy, myoepithelial cells could first be demonstrated in submandibular glands developing either in vitro or in vivo, at 21 days postconception. Contractions in the cultured rudiments began as early as 18 days postconception. Since neither smooth nor striated muscle could be identified in these glands by electron microscopy, the contractions are believed to result from myoepithelial activity that apparently may begin before ultrastructural evidence of myoepithelial differentiation is clearly present. Although, for over a century, myoepithelium has been presumed to have a contractile function and indirect evidence has lent ample support to this presumption, the present study represents the first direct cinematographic demonstration and characterization of myoepithelial contractions, under conditions in vitro.

Animals↗
PubMedDate not supplied

Biochemical and morphological studies of rat submandibular gland: II. Partial purification of proteins from granule-rich fraction.

Soluble proteins derived from a centrifuged and filtered granule-rich fraction of homogenized rat submandibular gland were analyzed by gel filtration, ion-exchange chromatography, and polyacrylamide gel electrophoresis. Both the granule-rich fraction and final supernatant fraction contained alkaline esterase activity. The major protein component, derived from granules of the convoluted tubules, was further resolved into a series of peptides ranging in molecular weight from 9,000 to 55,000 daltons.

Alkalies↗
PubMed1983

Mast cells in the submandibular gland of cat.

The effects of pilocarpine, compound 48/80 and the electrical stimulation of the chorda tympani nerve on the morphology of the mast cells from the submandibular gland of the cat were studied. All those different treatments of the gland caused the enlargement of the tissue mast cells. We observed also in the glands treated with pilocarpine and compound 48/80 a degranulation of mast cells. The degranulation was more expressed after the application of compound 48/80 to the animals.

Animals↗
PubMed1989

Permeability increase in rat submandibular gland by glandular substance P.

When the chorda tympani was stimulated at 20 Hz, infused horseradish peroxidase (HRP) entered the saliva through the tight junctions in the rat submandibular gland. This response was not inhibited by atropine but was reduced by a substance P(SP)-antagonist. The increase in junctional permeability for HRP never occurred in the gland that was stimulated at 5 Hz or in the SP-depleted gland. Our data suggest that the effect of parasympathetic stimulation on the increase in junctional permeability for macromolecules was due to the release of SP in the gland.

Animals↗
PubMed2003

Modulation of SOD activity in rat submandibular glands.

Reactive oxygen species (ROS) such as superoxide anion (O(2)(*-)), hydrogen peroxide (H(2)O(2)) and hydroxyl radical (*OH) are produced constantly in aerobic organisms. Submandibular glands of rats treated with isoproterenol (ISO) were analysed in respect to the level of superoxide dismutase (SOD), lipoperoxidation, protein oxidation and glutathione (GSH)/GSSG. Although ISO response is very well studied in salivary glands cells, the effect of ISO treatment on ROS productions or on SOD activation is not known. SOD activity increased after isoproterenol subcutaneous injection. In contrast, cycloheximide, a protein synthesis inhibitor, was found to decrease the total SOD activity. The GSH/GSSG ratio and the lipoperoxidation were also found to be increased after isoproterenol injection, while the protein oxidation was not affected by the same treatment. Injection of isoproterenol after cycloheximide treatment resulted in the recovery of the total SOD activity. This increase of activity is related to the amount of MnSOD isoform as shown by Western blot analysis. Our results suggest that the antioxidant protection of SOD may be activated during the salivary glands ISO treatment and that the mitochondrial MnSOD has an important role in the oxidative stress process.

Adrenergic beta-Agonists↗
PubMed1981

Effects of irradiation on the submandibular gland of the rat. An enzyme histochemical and ultrastructural study.

Single-dose cervical irradiation by cobalt 60 in rats induced lasting functional disturbances of the submandibular gland which were excessive when compared with the relative integrity of the gland as seen under the light microscope. Enzyme histochemical and ultrastructural studies revealed severe damage shortly after exposure with appearance of karyolytic bodies and autophagosomes accompanied by increased hydrolase activity. Mitochondrial alterations were concomitant with diminished ductal oxidative enzyme activity. Although most of these alterations resolved rapidly as a result of acinar and ductal cell repair and regeneration originating in the intercalated ducts, secretory abnormalities were still observed two months after exposure as evidenced by the accumulation of granules in acinar cells and the heterogeneity of ductal cell granules. These anomalies, comparable to those observed in sialadenoses, probably result from persistent alterations of intralobular nerve endings.

Animals↗
PubMed1977

Effect of neonatal sympathectomy on the postnatal differentiation of the submandibular gland of the rat.

Right superior cervical sympathectomy was performed in one-day old rats. This operation had a small but definite effect on the postnatal development of the submandibular gland. The gland on the sympathectomized side weighed less and contained smaller amounts of DNA, RNA and protein than the contralateral intact gland. The postnatal development of acinar cells and granular convoluted ductal cells was retarded in the sympathectomized gland. The acinar cells which differentiated after the ganglionectomy were smaller than those in the contralateral intact gland and were filled with secretory granules but devoid of basal basophilia. The rate of cellular proliferation in the sympathectomized gland was, however, similar to that intact gland at various ages studied.

Animals↗
PubMed1994

Mobilization of Ca2+ influx, but not of stored Ca2+, by extracellular ATP in rat submandibular gland acini.

To determine its effect on cytosolic Ca2+ levels (Ca2+i) and to assess its role as an autonomic cotransmitter, ATP was added to suspensions of rat pancreatic or submandibular gland acini loaded with fura-2. ATP had no effect on pancreatic acinar Ca2+i at either high (2.5 mM) or low (< 10 nM) extracellular Ca2+ concentrations (Ca2+o). In submandibular acini, ATP had little effect on Ca2+i when Ca2+o was < 10 nM but, with Ca2+o equal to 2.5 mM, ATP increased Ca2+i to a steady-state level 2-4 times the resting level of 50-100 nM. Addition of carbachol or epinephrine further increased Ca2+i. The mean and standard deviation of the ATP4- concentration at half of maximal activity (K0.5 ATP4-) was 33 +/- 7 microM and the Hill coefficient was 1.5 +/- 0.7 (n = 9). The active species is apparently ATP4-, as adding Mg2+ increases the total concentration of ATP needed to elevate Ca2+i. GTP, UTP, ADP and adenosine produced no significant changes in Ca2+i. However, 3'-O-(4-benzoyl)benzoyl ATP (BZATP) increased Ca2+i with a K0.5 BZATP of 2.4 +/- 0.5 microM and a Hill coefficient of 2.8 +/- 0.4 (n = 8). ATP-induced changes in Ca2+i were not due to a generalized increase in acinar-cell permeability and were unaffected by voltage-dependent Ca2+ channel blockers or by the presence of depolarizing concentrations of K+.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine↗