scholarly journals Altered Responses to Agonists after Chronic In Vivo Atropine Administration in Rat Parotid Acini

1993 ◽  
Vol 4 (3) ◽  
pp. 427-434 ◽  
Author(s):  
James E. Melvin ◽  
Guo H. Zhang
Keyword(s):  
Fluorescent Dyes ◽  
Inositol Phosphates ◽  
Chronic Administration ◽  
Fluid Secretion ◽  
Acute Response ◽  
Muscarinic Stimulation ◽  
Atropine Treatment ◽  
Rat Parotid ◽  
Efflux Pathway

Salivary gland hypofunction, resulting from a variety of perturbations including prescribed medications, is associated with adverse effects on the health of the oral cavity. In the present study, we investigated the in vivo effects of chronic administration of atropine, a muscarinic antagonist, on the acute response of rat parotid acini to a-adrenergic and muscarinic stimulation. The regulation of intracellular pH (pHi) and cytosolic free Ca2* ([Ca2+]i) were monitored using dual wavelength microfluorometry of the ion-sensitive fluorescent dyes, BCECF and fura-2, respectively. Chronic atropine treatment (40 mg/kg/d for 4 weeks) significantly increased the magnitude of the initial (<30 s) agonist-induced rise in [Ca2+]i, but did not alter the sustained increase in [Ca2+]i (>2 min). The generation of inositol trisphosphates and inositol tetrakisphosphates after 30 s of muscarinic stimulation was not significantly altered. The resting Cl- content, as well as the stimulated Cl- loss, were reduced in parotid acini after chronic atropine administration. In addition, the muscarinic- and a-adrenergic-induced intracellular acidification was blunted, suggesting that reduced HCO3- efflux occurs in acini isolated from atropine-treated animals. Our results indicate (1) that chronic atropine treatment does not inhibit the receptor-coupled generation of inositol phosphates or the resulting rise in [Ca2+]i and (2) chronic treatment may prevent the production of saliva either by reducing the driving force for anion-dependent fluid secretion or by preventing the activation of the anion efflux pathway.

scholarly journals Modulation of PKCδ tyrosine phosphorylation and activity in salivary and PC-12 cells by Src kinases

2001 ◽  
Vol 280 (6) ◽  
pp. C1498-C1510 ◽  
Author(s):  
Cyril Benes ◽  
Stephen P. Soltoff
Keyword(s):  
Substance P ◽  
Tyrosine Phosphorylation ◽  
Tyrosine Kinases ◽  
Tyrosine Phosphatase ◽  
Fluid Secretion ◽  
Pc 12 Cells ◽  
Intracellular Ca ◽  
Rat Parotid ◽  
Salivary Cell

Protein kinase C (PKC) δ becomes tyrosine phosphorylated in rat parotid acinar cells exposed to muscarinic and substance P receptor agonists, which initiate fluid secretion in this salivary cell. Here we examine the signaling components of PKCδ tyrosine phosphorylation and effects of phosphorylation on PKCδ activity. Carbachol- and substance P-promoted increases in PKCδ tyrosine phosphorylation were blocked by inhibiting phospholipase C (PLC) but not by blocking intracellular Ca2+ concentration elevation, suggesting that diacylglycerol, rather than d- myo-inositol 1,4,5-trisphosphate production, positively modulated this phosphorylation. Stimuli-dependent increases in PKCδ activity in parotid and PC-12 cells were blocked in vivo by inhibitors of Src tyrosine kinases. Dephosphorylation of tyrosine residues by PTP1B, a protein tyrosine phosphatase, reduced the enhanced PKCδ activity. Lipid cofactors modified the tyrosine phosphorylation-dependent PKCδ activation. Two PKCδ regulatory sites (Thr-505 and Ser-662) were constitutively phosphorylated in unstimulated parotid cells, and these phosphorylations were not altered by stimuli that increased PKCδ tyrosine phosphorylation. These results demonstrate that PKCδ activity is positively modulated by tyrosine phosphorylation in parotid and PC-12 cells and suggest that PLC-dependent effects of secretagogues on salivary cells involve Src-related kinases.

The Effect of Chronic Atropine Treatment on Salivary Composition and Caries in Rats

1989 ◽  
Vol 68 (12) ◽  
pp. 1739-1745 ◽  
Author(s):  
G.E. Watson ◽  
S.K. Pearson ◽  
J.L. Falany ◽  
D.J. Culp ◽  
L.A. Tabak ◽  
...  
Keyword(s):  
Chronic Administration ◽  
Cholinergic Receptor ◽  
Blood Levels ◽  
High Dose ◽  
Parotid Saliva ◽  
Sds Page ◽  
Atropine Treatment ◽  
High Doses

Many instances of salivary dysfunction in humans can be traced to the use of medications that have hyposalivary side-effects. In this study, atropine, a cholinergic antagonist, was administered chronically to rats by use of osmotic mini-pumps. Steady-state blood levels, similar to levels obtained in human multiple oral dosing, were thus maintained. Atropine delivered in this manner for 24 days was found to decrease protein concentration of parotid saliva (p<0.05) elicited by pilocarpine, and to increase smooth-surface caries scores (p<0.05) in rats fed a cariogenic diet. Parotid saliva collected via ductal cannulation from rats subjected to chronic atropine administration (and stimulated to secrete by pilocarpine) exhibited increased levels of two basic proline-rich proteins (Peak A and SP-3), as evaluated by SDS-PAGE, compared with those observed in saliva from controls. Cannulation of sublingual glands in animals receiving high doses of atropine produced no measurable secretion upon pilocarpine stimulation. Carbachol stimulation of dispersed cell aggregates of sublingual glands from sham-operated and high-dose atropine groups indicated that the glands responded similarly once the antagonist was washed from the system, implying that the lack of secretion in vivo was caused by antagonism of the cholinergic receptor by atropine. Our observations suggest that this model system can be exploited for determination of the effects of chronic administration of hyposalivary drugs on salivary composition and caries rates.

Ion transport systems in human labial acinar cells

1996 ◽  
Vol 270 (1) ◽  
pp. G213-G219 ◽  
Author(s):  
M. Paulais ◽  
I. H. Valdez ◽  
P. C. Fox ◽  
R. L. Evans ◽  
R. J. Turner
Keyword(s):  
Ion Transport ◽  
Fluid Secretion ◽  
Transport Systems ◽  
Muscarinic Agonist ◽  
Intracellular Acidification ◽  
Adrenergic Stimulation ◽  
Beta Adrenergic ◽  
Muscarinic Stimulation ◽  
Rat Parotid ◽  
Exchange Activity

Human labial acini were assayed for the presence of ion transport systems associated with salivary fluid secretion using microfluorometric methods. Na(+)-K(+)-Cl- contransport and Na+/H+ exchange activities (determined by their bumetanide and amiloride sensitivities, respectively) were found at levels approximately 50% of those seen in similarly assayed rat parotid acini, but little, if any, C1-/HCO3-exchange activity was observed. Also, when human labial acini were stimulated with the muscarinic agonist carbachol, little evidence of the intracellular acidification associated with HCO3- secretion by other salivary glands was found. Na+/H+ exchange activity in human labial acini was downregulated (approximately 40%) by beta-adrenergic stimulation and upregulated (approximately threefold) by muscarinic stimulation. In contrast, beta-adrenergic stimulation produced only a marginally significant increase in Na(+)-K(+)-Cl- cotransport activity, and muscarinic stimulation was without effect. We include that basolateral Na(+)-K(+)-Cl- cotransport appears to be the dominant mechanism driving Cl- secretion and thereby fluid secretion in this tissue.

Neuropeptides and Secretion

1987 ◽  
Vol 66 (2) ◽  
pp. 524-530 ◽  
Author(s):  
J. Ekström
Keyword(s):  
Secretory Granules ◽  
Fluid Secretion ◽  
Nerve Fibers ◽  
Parasympathetic Nerve ◽  
Calcitonin Gene ◽  
Rat Parotid Gland ◽  
Substance K ◽  
Rat Parotid ◽  
Stimulation Of

In the rat parotid gland, an atropine-resistant parasympathetic-nerve-evoked secretion was demonstrated in vivo. In the absence of atropine, the non-adrenergic, non-cholinergic transmitter release seemed to contribute to the fluid secretion and to be largely responsible for the secretion of amylase and acinar secretory granules. The gland was reached by nerve fibers containing substance P (SP), vasoactive intestinal peptide (VIP), and, to some extent, calcitonin-gene-related peptide (CGRP) via the parasympathetic auriculo-temporal nerve. Upon electrical stimulation of the nerve, these peptides were released. SP and substance K (SK), a novel tachykinin, induced a profuse watery secretion when injected i.v., while VIP caused a sparse but amylase-rich secretion. CGRP caused no secretion on its own. The tachykinin-evoked secretory response was enhanced by VIP and CGRP. A SPanalogue almost abolished the SP-evoked response, while the atropine-resistant parasympathetic response was only halved. None of the peptides under study can on its own account for the atropine-resistant parasympathetic secretion. The neuropeptides may play complementary roles in the regulation of the exocrine functions of the gland.

Optical imaging probes and their potential contribution to radiotracer development

2016 ◽  
Vol 55 (02) ◽  
pp. 51-62 ◽  
Author(s):  
S. Hermann ◽  
M. Schäfers ◽  
C. Höltke ◽  
A. Faust
Keyword(s):  
Optical Imaging ◽  
Fluorescent Dyes ◽  
Great Influence ◽  
Potential Contribution ◽  
Preclinical Evaluation ◽  
Guided Surgery ◽  
Fluorescence Guided Surgery ◽  
Imaging Probes ◽  
Unspecific Binding

SummaryOptical imaging has long been considered a method for histological or microscopic investigations. Over the last 15 years, however, this method was applied for preclinical molecular imaging and, just recently, was also able to show its principal potential for clinical applications (e.g. fluorescence-guided surgery). Reviewing the development and preclinical evaluation of new fluorescent dyes and target-specific dye conjugates, these often show characteristic patterns of their routes of excretion and biodistribution, which could also be interesting for the development and optimization of radiopharmaceuticals. Especially ionic charges show a great influence on biodistribution and netcharge and charge-distribution on a conjugate often determines unspecific binding or background signals in liver, kidney or intestine, and other organs.Learning from fluorescent probe behaviour in vivo and translating this knowledge to radio-pharmaceuticals might be useful to further optimize emerging and existing radiopharmaceuticals with respect to their biodistribution and thereby availability for binding to their targets.

scholarly journals Fibrillar pharmacology of functionalized nanocellulose

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Sam Wong ◽  
Simone Alidori ◽  
Barbara P. Mello ◽  
Bryan Aristega Almeida ◽  
David Ulmert ◽  
...  
Keyword(s):  
Aspect Ratio ◽  
Fluorescent Dyes ◽  
Pharmacokinetic Profile ◽  
Water Soluble ◽  
Size Exclusion ◽  
Hydroxyl Groups ◽  
Target Tissue ◽  
Exclusion Chromatography ◽  
Specific Accumulation

AbstractCellulose nanocrystals (CNC) are linear organic nanomaterials derived from an abundant naturally occurring biopolymer resource. Strategic modification of the primary and secondary hydroxyl groups on the CNC introduces amine and iodine group substitution, respectively. The amine groups (0.285 mmol of amine per gram of functionalized CNC (fCNC)) are further reacted with radiometal loaded-chelates or fluorescent dyes as tracers to evaluate the pharmacokinetic profile of the fCNC in vivo. In this way, these nanoscale macromolecules can be covalently functionalized and yield water-soluble and biocompatible fibrillar nanoplatforms for gene, drug and radionuclide delivery in vivo. Transmission electron microscopy of fCNC reveals a length of 162.4 ± 16.3 nm, diameter of 11.2 ± 1.52 nm and aspect ratio of 16.4 ± 1.94 per particle (mean ± SEM) and is confirmed using atomic force microscopy. Size exclusion chromatography of macromolecular fCNC describes a fibrillar molecular behavior as evidenced by retention times typical of late eluting small molecules and functionalized carbon nanotubes. In vivo, greater than 50% of intravenously injected radiolabeled fCNC is excreted in the urine within 1 h post administration and is consistent with the pharmacological profile observed for other rigid, high aspect ratio macromolecules. Tissue distribution of fCNC shows accumulation in kidneys, liver, and spleen (14.6 ± 6.0; 6.1 ± 2.6; and 7.7 ± 1.4% of the injected activity per gram of tissue, respectively) at 72 h post-administration. Confocal fluorescence microscopy reveals cell-specific accumulation in these target tissue sinks. In summary, our findings suggest that functionalized nanocellulose can be used as a potential drug delivery platform for the kidneys.

Involvement of αENaC and Nedd4-2 in the conversion from lung fluid secretion to fluid absorption at birth in the rat as assayed by RNA interference analysis

2007 ◽  
Vol 293 (4) ◽  
pp. L1069-L1078 ◽  
Author(s):  
Tianbo Li ◽  
Shyny Koshy ◽  
Hans G. Folkesson
Keyword(s):  
Extravascular Lung Water ◽  
Fluid Secretion ◽  
Alveolar Epithelial Cells ◽  
Na Channel ◽  
Fluid Absorption ◽  
Lung Water ◽  
Fourfold Increase ◽  
Lung Fluid ◽  
Near Term

To explore interactions between the epithelial Na channel (ENaC) and neural precursor expressed, developmentally downregulated protein 4-2 (Nedd4-2) at the conversion of the rat lung from fluid secretion to absorption at birth, we used small-interfering RNA (siRNA) against αENaC and Nedd4-2. siRNA-generating plasmid DNA (pDNA) was administered via trans-thoracic intrapulmonary (ttip) injection 24 h before ENaC and Nedd4-2 expression, extravascular lung water, and mortality were measured. αENaC mRNA and protein were specifically reduced by ∼65% after pSi-4 injection. Nedd4-2 mRNA and protein were reduced by ∼60% after pSi-N1 injection. Interestingly, αENaC and βENaC mRNA and protein expression were increased after Nedd4-2 silencing. Extravascular lung water was significantly increased after αENaC silencing and reduced after Nedd4-2 silencing. αENaC silencing resulted in a fourfold increase in newborn mortality, whereas silencing Nedd4-2 did not affect mortality. We also isolated distal lung epithelial (DLE) cells after in vivo αENaC or Nedd4-2 silencing and measured αENaC or Nedd4-2 expression in freshly isolated DLE cells. In these DLE cells, there were attenuated αENaC or Nedd4-2 mRNA and protein, thus demonstrating that αENaC and Nedd4-2 silencing occurred in alveolar epithelial cells after ttip injection. We also looked for pDNA by PCR to determine pDNA presence in the lungs and found strong evidence for pDNA presence in both lungs. Thus we provide evidence that ENaC and Nedd4-2 are involved in the transition from lung fluid secretion to fluid absorption near term and at birth.

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