scholarly journals Effects of muscarinic, alpha-adrenergic, and substance P agonists and ionomycin on ion transport mechanisms in the rat parotid acinar cell. The dependence of ion transport on intracellular calcium.

1989 ◽  
Vol 93 (2) ◽  
pp. 285-319 ◽  
Author(s):  
S P Soltoff ◽  
M K McMillian ◽  
L C Cantley ◽  
E J Cragoe ◽  
B R Talamo
Keyword(s):  
Substance P ◽  
Ion Transport ◽  
Acinar Cell ◽  
Fluid Secretion ◽  
Ion Fluxes ◽  
Transport Systems ◽  
K Channel ◽  
Muscarinic Agonist ◽  
Parotid Acinar Cell ◽  
Rat Parotid

The relationship between receptor-mediated increases in the intracellular free calcium concentration [( Ca]i) and the stimulation of ion fluxes involved in fluid secretion was examined in the rat parotid acinar cell. Agonist-induced increases in [Ca]i caused the rapid net loss of up to 50-60% of the total content of intracellular chloride (Cli) and potassium (Ki), which is consistent with the activation of calcium-sensitive chloride and potassium channels. These ion movements were accompanied by a 25% reduction in the intracellular volume. The relative magnitudes of the losses of Ki and the net potassium fluxes promoted by carbachol (a muscarinic agonist), phenylephrine (an alpha-adrenergic agonist), and substance P were very similar to their characteristic effects on elevating [Ca]i. Carbachol stimulated the loss of Ki through multiple efflux pathways, including the large-conductance Ca-activated K channel. Carbachol and substance P increased the levels of intracellular sodium (Nai) to more than 2.5 times the normal level by stimulating the net uptake of sodium through multiple pathways; Na-K-2Cl cotransport accounted for greater than 50% of the influx, and approximately 20% was via Na-H exchange, which led to a net alkalinization of the cells. Ionomycin stimulated similar fluxes through these two pathways, but also promoted sodium influx through an additional pathway which was nearly equivalent in magnitude to the combined uptake through the other two pathways. The carbachol-induced increase in Nai and decrease in Ki stimulated the activity of the sodium pump, measured by the ouabain-sensitive rate of oxygen consumption, to nearly maximal levels. In the absence of extracellular calcium or in cells loaded with the calcium chelator BAPTA (bis[o-aminophenoxy]ethane-N,N,N',N'-tetraacetic acid) the magnitudes of agonist- or ionomycin-stimulated ion fluxes were greatly reduced. The parotid cells displayed a marked desensitization to substance P; within 10 min the elevation of [Ca]i and alterations in Ki, Nai, and cell volume spontaneously returned to near baseline levels. In addition to quantitating the activation of various ion flux pathways in the rat parotid acinar cell, these results demonstrate that the activation of ion transport systems responsible for fluid secretion in this tissue is closely linked to the elevation of [Ca]i.

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.

scholarly journals Effects of extracellular ATP on ion transport systems and [Ca2+]i in rat parotid acinar cells. Comparison with the muscarinic agonist carbachol.

1990 ◽  
Vol 95 (2) ◽  
pp. 319-346 ◽  
Author(s):  
S P Soltoff ◽  
M K McMillian ◽  
E J Cragoe ◽  
L C Cantley ◽  
B R Talamo
Keyword(s):  
Relative Proportion ◽  
Acinar Cells ◽  
Initial Response ◽  
Extracellular Atp ◽  
Cation Channel ◽  
Transport Systems ◽  
K Channel ◽  
Muscarinic Agonist ◽  
Parotid Acinar Cells ◽  
Rat Parotid

The effects of extracellular ATP on ion fluxes and the intracellular free Ca2+ concentration ([Ca2+]i) were examined using a suspension of rat parotid acinar cells and were contrasted with the effects of the muscarinic agonist carbachol. Although ATP and carbachol both rapidly increased [Ca2+]i about threefold above the resting level (200-250 nM), the effect of ATP was due primarily to an influx of Ca2+ across the plasma membrane, while the initial response to carbachol was due to a release of Ca2+ from intracellular stores. Within 10 s, ATP (1 mM) and carbachol (20 microM) reduced the cellular Cl- content by 39-50% and cell volume by 15-25%. Both stimuli reduced the cytosolic K+ content by 57-65%, but there were marked differences in the rate and pattern of net K+ movement as well as the effects of K+ channel inhibitors on the effluxes initiated by the two stimuli. The maximum rate of the ATP-stimulated K+ efflux (approximately 2,200 nmol K+/mg protein per min) was about two-thirds that of the carbachol-initiated efflux rate, and was reduced by approximately 30% (vs. 60% for the carbachol-stimulated K+ efflux) by TEA (tetraethylammonium), an inhibitor of the large conductance (BK) K+ channel. Charybdotoxin, another K+ channel blocker, was markedly more effective than TEA on the effects of both agonists, and reduced the rate of K+ efflux initiated by both ATP and carbachol by approximately 80%. The removal of extracellular Ca2+ reduced the ATP- and the carbachol-stimulated rates of K+ efflux by 55 and 17%, respectively. The rate of K+ efflux initiated by either agonist was reduced by 78-95% in cells that were loaded with BAPTA to slow the elevation of [Ca2+]i. These results indicated that ATP and carbachol stimulated the efflux of K+ through multiple types of K(+)-permeable channels, and demonstrated that the relative proportion of efflux through the different pathways was different for the two stimuli. ATP and carbachol also stimulated the rapid entry of Na+ into the parotid cell, and elevated the intracellular Na+ content to 4.4 and 2.6 times the normal level, respectively. The rate of Na+ entry through Na(+)-K(+)-2Cl- cotransport and Na(+)-H+ exchange was similar whether stimulated by ATP, carbachol, or ionomycin, and uptake through these two carrier-mediated transporters accounted for 50% of the ATP-promoted Na+ influx. The remainder may be due to a nonselective cation channel and an ATP-gated cation channel that is also permeable to Ca2+.(ABSTRACT TRUNCATED AT 400 WORDS)

Ion transport mechanisms in rat parotid intralobular striated ducts

1994 ◽  
Vol 266 (6) ◽  
pp. C1594-C1602 ◽  
Author(s):  
M. Paulais ◽  
E. J. Cragoe ◽  
R. J. Turner
Keyword(s):  
Ion Transport ◽  
Channel Blocker ◽  
K Channel ◽  
Muscarinic Agonist ◽  
Transport Mechanisms ◽  
Adrenergic Agonist ◽  
Acid Load ◽  
Rat Parotid ◽  
H Channels ◽  
Beta Adrenergic Agonist

The intracellular pH (pHi) indicator 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein and microfluorimetry were used to characterize several ion transport mechanisms in rat parotid striated ducts. The recovery of ductal pHi from an acute acid load was Na+ dependent and inhibited by the amiloride analogue ethylisopropylamiloride with 50% inhibitory concentration 4.7 +/- 0.8 microM, indicating the presence of a Na(+)-H+ exchanger of the amiloride-insensitive type. The rate of this recovery was stimulated approximately 20% in ducts pretreated with the muscarinic agonist carbachol (10(-5) M) and inhibited approximately 20% in ducts pretreated with the beta-adrenergic agonist isoproterenol (10(-6) M). Upon removal of extracellular K+, ductal pHi rapidly decreased (0.19 +/- 0.02 pH units/min), consistent with a coupling between K+ and H+ (or OH-) fluxes in this tissue. In HCO(3-)-containing medium, the acidification due to K+ removal was blunted, arguing against ductal K(+)-HCO3- cotransport. However, the effect of K+ removal was inhibited by the K+ channel blocker Ba2+ (1 mM) and by the H+ channel blocker Zn2+ (25 microM), consistent with the involvement of electrically coupled K+ and H+ channels. The effect of K+ removal was unaffected by pretreatment of ducts with isoproterenol (10(-6) M) but markedly inhibited (approximately 50%) by pretreatment with carbachol (10(-5) M). No evidence for a significant component of Cl(-)-HCO3- exchange was found in striated ducts. The properties of the Na(+)-H+ exchanger and K(+)-H+ exchange mechanism identified here are consistent with their involvement in ductal salt reabsorption and secretion.

Secretagogue-induced 86Rb+ efflux from bovine parotid is HCO3- dependent

1993 ◽  
Vol 264 (1) ◽  
pp. R162-R168 ◽  
Author(s):  
S. I. Lee ◽  
R. J. Turner
Keyword(s):  
Anion Channel ◽  
Fluid Secretion ◽  
K Channel ◽  
Channel Blockers ◽  
Muscarinic Agonist ◽  
Electrically Conductive ◽  
Significant Evidence ◽  
Anion Secretion ◽  
Conductive Pathway ◽  
Rat Parotid

Muscarinic agonist (carbachol)-induced K+ loss from a bovine parotid mince was studied using 86Rb+ as a K+ marker. In contrast to our previous studies with the rat parotid [Am. J. Physiol. 261 (Gastrointest. Liver Physiol. 24): G111-G118, 1991] in which both Cl(-)-dependent and HCO3(-)-dependent components of carbachol-induced 86Rb+ efflux were observed, no significant evidence for Cl(-)-dependent 86Rb+ loss was detected in the bovine parotid. HCO3(-)-dependent agonist-induced 86Rb+ loss was blunted by K+ and Cl- channel blockers and by removal of extracellular Ca2+, consistent with the hypothesis that this 86Rb+ loss occurs via a Ca(2+)-activated K+ channel and that this cation loss serves to electrically balance the concomitant loss of HCO3- via an electrically conductive pathway, presumably an apical anion channel. Acetate, formate, and propionate could substitute for HCO3-. Interpreted in terms of current models of salivary fluid secretion, which hypothesize that the production of fluid is secondary to anion secretion accompanied by an electrically coupled K+ loss, these results indicate that salivary production in the bovine parotid is driven almost exclusively by acinar HCO3- secretion.

Cl- fluxes related to fluid secretion by the rat parotid: involvement of Cl(-)-HCO3- exchange

1992 ◽  
Vol 262 (3) ◽  
pp. G393-G398 ◽  
Author(s):  
J. E. Melvin ◽  
R. J. Turner
Keyword(s):  
Channel Blocker ◽  
Kinetic Studies ◽  
Fluid Secretion ◽  
Disulfonic Acid ◽  
K Channel ◽  
Maximal Effect ◽  
Partial Recovery ◽  
Free Medium ◽  
Cl Channels ◽  
Rat Parotid

Muscarinic-induced 36Cl- and 86Rb+ (K+ substitute) fluxes were studied in rat parotid acini. Stimulation resulted in a rapid [half time (t1/2) less than 30 s] decrease in both Cl- and Rb+ content (approximately 50 and 30%, respectively) followed by a slower partial recovery (t1/2 approximately 3-4 min) to approximately 80% of resting levels for both ions. Cl- loss was inhibited by the venom of Leiurus quinquestriatus, which contains the maxi-K+ channel blocker charybdotoxin. Cl- recovery was blunted in the presence of bumetanide, an inhibitor of Na(+)-K(+)-Cl- cotransport, or on HCO3- removal and was completely blocked in the presence of bumetanide and 4,4' diisothiocyanostilbene-2,2' disulfonic acid (DIDS), an inhibitor of Cl(-)-HCO3- exchange. In HCO3(-)-containing medium a rapid (t1/2 less than 1 min), DIDS-inhibitable cytoplasmic alkalinization (approximately 0.4 pH unit) was observed in acini switched to a Cl(-)-free solution. This alkalinization was not seen in HCO3(-)-free medium but persisted in the absence of Na+, consistent with the presence of a potent Na(+)-independent Cl(-)-HCO3- exchanger. Kinetic studies indicated that the half-maximal effect of this exchanger for extracellular Cl- was approximately 18 mM. These results are consistent with the hypothesis that secretagogue-induced KCl loss by salivary acinar cells occurs via electrically coupled K+ and Cl- channels. In addition, they provide strong evidence that Cl- entry into, and thus fluid secretion by, these cells is mediated by both Cl(-)-HCO3- exchange and Na(+)-K(+)-Cl- cotransport.

scholarly journals Interaction between zymogen granule and membrane of rat parotid acinar cell, in vitro.

1993 ◽  
Vol 61 ◽  
pp. 57
Author(s):  
Takahiro Nagao ◽  
Ritsuko Fujimoto ◽  
Hideaki Nishio ◽  
Fumiaki Hata
Keyword(s):  
Acinar Cell ◽  
Zymogen Granule ◽  
Parotid Acinar Cell ◽  
Rat Parotid

Functional and molecular characterization of the fluid secretion mechanism in human parotid acinar cells

2007 ◽  
Vol 292 (6) ◽  
pp. R2380-R2390 ◽  
Author(s):  
Tetsuji Nakamoto ◽  
Alaka Srivastava ◽  
Victor G. Romanenko ◽  
Catherine E. Ovitt ◽  
Patricia Perez-Cornejo ◽  
...  
Keyword(s):  
Salivary Gland ◽  
Ion Transport ◽  
Acinar Cells ◽  
Fluid Secretion ◽  
K Channel ◽  
Resting Membrane Potential ◽  
Specific Cell ◽  
Parotid Glands ◽  
Voltage Dependent ◽  
Intracellular Ca

The strategies available for treating salivary gland hypofunction are limited because relatively little is known about the secretion process in humans. An initial microarray screen detected ion transport proteins generally accepted to be critically involved in salivation. We tested for the activity of some of these proteins, as well as for specific cell properties required to support fluid secretion. The resting membrane potential of human acinar cells was near −51 mV, while the intracellular [Cl−] was ∼62 mM, about fourfold higher than expected if Cl ions were passively distributed. Active Cl− uptake mechanisms included a bumetanide-sensitive Na+-K+-2Cl− cotransporter and paired DIDS-sensitive Cl−/HCO3− and EIPA-sensitive Na+/H+ exchangers that correlated with expression of NKCC1, AE2, and NHE1 transcripts, respectively. Intracellular Ca2+ stimulated a niflumic acid-sensitive Cl− current with properties similar to the Ca2+-gated Cl channel BEST2. In addition, intracellular Ca2+ stimulated a paxilline-sensitive and voltage-dependent, large-conductance K channel and a clotrimazole-sensitive, intermediate-conductance K channel, consistent with the detection of transcripts for KCNMA1 and KCNN4, respectively. Our results demonstrate that the ion transport mechanisms in human parotid glands are equivalent to those in the mouse, confirming that animal models provide valuable systems for testing therapies to prevent salivary gland dysfunction.

Morphological Effects of WR-2721 on the Rat Parotid Acinar Cell

1978 ◽  
Vol 75 (2) ◽  
pp. 327 ◽  
Author(s):  
N. E. Pratt ◽  
M. Sodicoff
Keyword(s):  
Acinar Cell ◽  
Parotid Acinar Cell ◽  
Rat Parotid

scholarly journals A mathematical model of fluid secretion from a parotid acinar cell

2007 ◽  
Vol 248 (1) ◽  
pp. 64-80 ◽  
Author(s):  
Elan Gin ◽  
Edmund J. Crampin ◽  
David A. Brown ◽  
Trevor J. Shuttleworth ◽  
David I. Yule ◽  
...  
Keyword(s):  
Mathematical Model ◽  
Acinar Cell ◽  
Fluid Secretion ◽  
Parotid Acinar Cell
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