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Biomedical subjects

J R Garrett

Publications and source records attributed to J R Garrett.

At least 73 records · Page 4Linked to original sources

Increase of microliths in inactive salivary glands of cat.

Secretory inactivity could be a factor in the formation of microliths, and so their occurrence in feline salivary glands after the secretory inactivity produced by parasympathectomy was investigated. Parasympathectomy was followed by a greatly increased occurrence of microliths in the submandibular salivary gland, but not in the parotid and sublingual, which may relate to residual secretory activity in these glands. This discovery suggests that secretory inactivity may indeed be a factor in the production of microliths in human salivary glands, and consequently of chronic sialadenitis and sialothiasis.

Animals↗

Secretory oedema in diabetic submandibular glands during parasympathetic nerve stimulation: relationship to microvascular abnormalities in streptozotocin-treated rats.

1. Submandibular secretion during parasympathetic stimulation (5 Hz) was examined in streptozotocin-diabetic and age-matched control rats. 2. At 3 weeks, but not 3 and 6 months, flow rate was initially greater than in controls, but it declined rapidly after 30 min. 3. The reduction in flow rate was associated with oedema of the gland. 4. At 3 months, graded stimulation revealed a tendency to oedema at frequencies of 10 Hz and above. 5. Morphologically, submandibular capillary density was increased in diabetic rats. 6. Thus, in diabetes the submandibular gland appears less able to withstand continuous parasympathetic stimulation, due in part to an increase in tissue capillary area.

Animals↗

Endothelium-derived vasodilator responses to sympathetic stimulation of the submandibular gland in the cat.

1. The extent to which vasodilator responses to electrical stimulation of the sympathetic innervation, in the submandibular gland of the cat, depend upon release of endothelium-derived relaxing factor (EDRF) within the gland has been investigated in anaesthetized cats given N-nitro-L-arginine methyl ester (L-NAME) which specifically blocks the synthesis of EDRF from arginine. 2. Close intra-arterial infusions of L-NAME (> or = 100 mg kg-1) produced a steady and significant rise in mean aortic pressure together with a steady increase in basal submandibular vascular resistance over the next 20-30 min. It also reduced, but failed to abolish, the vasodilatation which occurs during intermittent stimulation of the sympathetic innervation (20 Hz for 1 s at 10 s intervals) together with the after-dilatation which occurs immediately after a period of continuous stimulation of these nerve fibres (2 Hz). 3. In cats pretreated with the beta-blocker propranolol (> or = 1.0 mg kg-1) both vasodilator responses were reduced, but persisted until L-NAME was administered, whereupon both were abolished. 4. It is concluded that release of EDRF within the submandibular gland of the cat contributes to the basal tone of the vasculature and is responsible for the alpha-adrenergic vasodilator responses to stimulation of the sympathetic innervation, but not for the beta-adrenergic vasodilator responses.

Animals↗

Nerve-induced secretion of glycoconjugates from cat submandibular glands: a correlative study with lectin probes on tissues and saliva.

Horseradish peroxidase-conjugated lectins were used on tissue sections to localize the main secretory glycoproteins in cat submandibular glands and on Western blots to evaluate their movement into saliva with selective nerve stimulation. Central acinar cells bound lectins from Arachis hypogaea (PNA) specific for the terminal disaccharide Gal beta 1, 3GalNac, Griffonia simplifolia (GSA I-B4) specific for terminal alpha Gal, and Lotus tetragonolobus (LTA) specific for fucose. Lectins from Limax flavus (LFA) specific for sialic acid and Dolichos biflorus (DBA) specific for terminal alpha GalNac reacted preferentially with demilunar cells, whereas apical granules in striated ducts were recognized principally by LTA. Parasympathetic stimulation promoted the release of lectin-reactive glycoconjugates from both central and demilunar cells. In contrast, sympathetic stimulation caused almost complete release of LTA-reactive granules in striated ducts and only moderate secretion from demilunar cells. Lectin blots of stimulated saliva discriminated many of the constituent bands, providing information about their glycosylation. Several bands were common to both parasympathetic and sympathetic saliva, and many bands gave wider ranges of lectin binding than anticipated from the histochemistry. The major component in parasympathetic saliva was a glycoconjugate of less than 12 KD which reacted with every lectin tested. Lectin blots of sympathetic saliva showed a prominent diffuse LTA-reactive band around 33 KD, which was attributed to tissue kallikrein. The identity and cellular origin of most bands in stimulated submandibular saliva are still unclear but the technique shows considerable promise for improving the recognition and characterization of individual glycoconjugates.

Animals↗

Secretory responses in granular ducts and acini of submandibular glands in vivo to parasympathetic or sympathetic nerve stimulation in rats.

The roles of sympathetic and parasympathetic nerves in the secretion of saliva from submandibular glands of rats have been tested by electrical stimulation of either nerve for 1 h unilaterally in separate animals. The flows of saliva thereby induced and their protein content were monitored. Structural changes in each gland were assessed by light- and electron microscopy and compared with the unstimulated contralateral control gland, and the extent of the changes was determined morphometrically. Sympathetic nerve stimulation induced a relatively low flow of saliva that was rich in protein and was accompanied by extensive degranulation from both acinar and granular duct cells. In contrast parasympathetic nerve stimulation induced a considerable flow of saliva that had a low protein content and no detectable degranulation occurred from the secretory cells. It is possible, therefore, that some protein in parasympathetic saliva may have arisen from a non-granular pathway.

Animals↗

Rat sublingual salivary glands: secretory changes on parasympathetic or sympathetic nerve stimulation and a reappraisal of the adrenergic innervation of striated ducts.

Sublingual glands were examined by light and electron microscopy after stimulating the parasympathetic nerve (5 Hz continuously) or the sympathetic nerve (50 Hz in bursts 1 s every 10 s) and compared with contralateral, unstimulated, normal glands from each animal. Parasympathetic stimulation caused secretion of mucin from the tubulo-acini and possibly a small amount of degranulation from the demilunes but no changes were detected in the striated ducts. Sympathetic stimulation, on the other hand, had no effect on the tubulo-acini or demilunes but caused a surprisingly extensive degranulation of the striated ducts plus loss of glycogen from their cells. Reassessment of the adrenergic innervation in the glands was therefore undertaken, by means of catecholamine fluorescence. This identified a regular association between adrenergic nerves and the striated ducts not only in sublingual but also in submandibular glands; features that have not previously been recognized. There was, however, only a sparse adrenergic innervation of the other parenchymal elements in the sublingual glands.

Animals↗

Proteinase activities in bovine atrium and the possible role of mast cell tryptase in the processing of atrial natriuretic factor (ANF).

1. Using low salt, Triton X-100 and high salt extracts of bovine atria, two main proteinases were identified by means of fluorogenic oligopeptide substrates. 2. An acidic proteinase, extracted in low salt and Triton X-100 was identified as cathepsin B, but it caused little hydrolysis of the Z-Gly-Pro-Arg- containing substrate that resembles the cleavage site for activation of pro-ANF. 3. An alkaline proteinase was extracted only with high salt and had characteristics of the serine proteinase tryptase. It cleaved Z-Gly-Pro-Arg- containing substrates more efficiently than others tested and was localized in and around mast cells histochemically. Previously, Imada et al., 1988 (J. biol. Chem. 263, 9515-9519) found an identical enzyme would cleave ANF from pro-ANF. 4. These results suggest therefore that mast cell tryptase may be involved in the activation of ANF from pro-ANF.

Amino Acid Sequence↗

Serum amylase of non-parotid and non-pancreatic origin increases on feeding in rats and may originate from the liver.

1. In order to evaluate possible non-salivary contributions to the content of salivary-type amylase in the circulation, parotid glands--the only salivary source of amylase in rats--have been totally removed and the effects on serum amylase have been assessed, after fasting and at different times after feeding. 2. Despite the parotidectomy the resting level of salivary-type amylase remained the same and an increase was still found to occur on feeding. 3. Isoelectric focusing has identified additional isoforms of amylase in serum and liver distinct from those occurring in parotid saliva. 4. The liver therefore may be contributing to the fasting levels of serum amylase and to the increases that occur on feeding, in rats.

Amylases↗

Morphological effects of sympathetic nerve stimulation on rat parotid glands 3-4 weeks after the induction of streptozotocin diabetes.

Male Wistar rats were fasted overnight and anaesthetized 3-4 weeks after the induction of streptozotocin diabetes. The right parotid ducts were cannulated, and parotid salivary flow was induced by stimulating the sympathetic trunk in bursts (50 Hz, 1:10). Stimulated and unstimulated glands were weighed, fixed for morphometric analysis, and assayed for total protein and amylase. Gland weights did not differ between diabetic and control rats. Nevertheless, total protein (6.17 +/- 5.40 mg) and amylase (1.32 +/- 0.49 mg) output from the gland were reduced in diabetic animals compared with controls (13.73 +/- 2.81 and 3.41 +/- 0.51 mg, respectively). Morphometric analysis of unstimulated glands showed no differences in either acinar cell profile area, or in the number of secretory granule profiles/cell in unstimulated glands. Upon sympathetic stimulation, however, the depletion of granule profiles was significantly less in diabetic than in control rats. Finally, the mean diameter of secretory granules was significantly less in diabetic (0.89 +/- 0.05 microM) than in control (1.04 +/- 0.06 microM) glands.

Amylases↗

Effects of parasympathectomy on protein composition of sympathetically evoked parotid saliva in rats.

1. Male Wistar rats were given unilateral postganglionic parasympathectomies by sectioning the auriculo-temporal nerve. 2. Analyses of the protein compositions of sympathetically induced saliva from both glands 1 week later revealed changes in the proportions of different secretory proteins, in particular amylase and basic proline-rich proteins were decreased. 3. These results suggest that parasympathetic impulses are required for the normal synthesis of amylase by parotid parenchyma in rats. Basic proline-rich proteins, known to require a sympathetic drive for normal synthesis, appear to require a parasympathetic input as well.

Animals↗

Factors influencing the movement of parotid amylase into the serum of rats on feeding.

Feeding hard chow to rats for 1 h caused an increase in parotid-type amylase in the serum. The increase was largely inhibited by parasympathetic or sympathetic denervation of the parotid glands and also by beta-adrenoceptor blockade. The results suggest that movement of amylase into the blood is influenced not only by the volume of saliva being formed but also by its amylase content.

Amylases↗

Na+, K(+)-ATPase in cat salivary glands and changes induced by nerve stimulation: an immunohistochemical study.

The enzyme Na+, K(+)-ATPase was localized immunohistochemically in major salivary glands of the cat before and after autonomic nerve stimulation. Immunostaining was limited to basolateral plasma membranes. Cells lining striated and excretory ducts contained abundant Na+, K(+)-ATPase and showed no changes with neural stimulation. Serous-type cells in resting glands varied in reactivity, showing weak to moderate staining intensity in the parotid gland and more uniform staining of greater intensity in the sublingual gland. In contrast, demilune cells in the resting submandibular gland showed little if any staining. Mucous-type cells were negative in all glands. Parasympathetic stimulation promoted a gradual increase in immunostaining of submandibular demilune cells, which became marked with time. Sympathetic stimulation produced no detectable changes in Na+, K(+)-ATPase immunoreactivity in any site. These results support the concept that basolateral Na+, K(+)-ATPase is essential to the formation of a near-isotonic primary saliva by serous-type cells. The mechanism whereby parasympathetic stimulation evokes a marked flow of submandibular saliva remains unexplained, but has now been shown to involve a marked increase in the immunoreactivity of Na+, K(+)-ATPase at the base of the gland's demilune cells.

Animals↗

Effects of inhibitors on proteinase activities in gels following isoelectric focussing.

Homogenates of rat submandibular glands were electrophoresed on isoelectric focussing gels and bands of proteinase activity demonstrated by means of overlay membranes impregnated with fluorogenic oligopeptide substrates. Inhibition characteristics of individual proteinase bands were tested within the gels and after excision and subsequent elution from the gels. Some bands of activity were resistant to inhibition by pre-incubation of the gel with certain inhibitors, but were subsequently found to be inhibited when tested with the same substrate after elution. These inconsistencies were influenced by the use of different substrates. The results indicate that, though some useful screening information may be obtained by applying inhibitors to the gels, this system does not permit a reliable assessment of inhibition characteristics which require subsequent elution and assay.

Amino Acid Sequence↗

Nerve-induced secretion of parotid acinar granules in cats.

Electron microscopy of cat parotid glands revealed great heterogeneity in the secretory granules of normal unstimulated acinar cells. Electrical stimulation of the parasympathetic nerve to the gland evoked a copious flow of parotid saliva which was accompanied by an extensive depletion of the secretory granules from the acinar cells. Exocytosis was captured as it was occurring by means of perfusion-fixation, and showed that the events occur in a conventional manner. Stimulation of the sympathetic nerve caused only a very small flow of saliva, and no acinar degranulation was detected. It can be concluded that the parasympathetic secretomotor axons provide the main drive for parotid acinar degranulation in the cat. This contrasts with the rat in which sympathetic impulses provide the main stimulus for parotid acinar degranulation. These dissimilarities serve to emphasise how extensively species differences may influence autonomic responses in salivary glands.

Animals↗