ATP activates a cation-permeable pathway in rat parotid acinar cells

1992 ◽  
Vol 262 (4) ◽  
pp. C934-C940 ◽  
Author(s):  
S. P. Soltoff ◽  
M. K. McMillian ◽  
B. R. Talamo
Keyword(s):  
Membrane Potential ◽  
Uptake Rate ◽  
Purinergic Receptor ◽  
Acinar Cells ◽  
Muscarinic Agonist ◽  
Tetraacetic Acid ◽  
Nucleotide Analogues ◽  
Parotid Acinar Cells ◽  
Biphasic Effect ◽  
Rat Parotid

Effects of several purinergic receptor agonists were examined on rat parotid acinar cells. Extracellular ATP stimulated 45Ca2+ uptake into isolated rat parotid acinar cells in a concentration-dependent fashion (EC50 approximately 125 microM ATP) at a maximum rate of approximately 6 nmol.mg protein-1.min-1. In the absence of extracellular Na+, ATP increased the uptake rate by greater than 100%. Increasing concentrations of extracellular Na+ reduced the ATP-stimulated rate of 45Ca2+ entry in a graded fashion (IC50 16.6 mM), suggesting that Ca2+ and Na+ compete for entry. Uptake rate was not reduced when intracellular Ca2+ was buffered with 1,2-bis(2-aminophenoxy)ethane-N,N,N'N'-tetraacetic acid, indicating that the effects of ATP were not initiated by an elevation in intracellular free Ca2+ concentration. 3-O-(4'-benzoyl)benzoyl-ATP was much more potent (EC50 approximately 4 microM) and stimulated Ca2+ influx at a greater rate (approximately 12 nmol.mg protein-1.min-1) than ATP. Other nucleotide analogues, including adenosine 5'-O-(3-thiotriphosphate), 2-methylthio-ATP, and 5'-adenylylimidodiphosphate, were much less effective than ATP. ATP produced a biphasic effect on membrane potential: an initial hyperpolarization was followed by a rapid depolarization. The depolarization was greatly reduced in the absence of extracellular Na+, but not in the absence of extracellular Ca2+, indicating that the majority of the depolarizing current was due to Na+ entry. Effects of ATP on the membrane potential were distinguishable from those of the Ca2+ ionophore ionomycin and the muscarinic agonist carbachol. Depolarization of the cells by gramicidin or K+ did not produce an increase in 45Ca2+ uptake.(ABSTRACT TRUNCATED AT 250 WORDS)

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)

Mechanism of secretagogue-induced HCO3- and Cl- loss from rat parotid acini

1991 ◽  
Vol 261 (1) ◽  
pp. G111-G118 ◽  
Author(s):  
S. I. Lee ◽  
R. J. Turner
Keyword(s):  
Acinar Cells ◽  
Time Dependent ◽  
K Channel ◽  
Channel Blockers ◽  
Muscarinic Agonist ◽  
Transport Pathways ◽  
Parotid Acinar Cells ◽  
Muscarinic Response ◽  
Muscarinic Stimulation ◽  
Rat Parotid

The Cl(-)- and HCO3(-)-dependent components of muscarinic agonist (carbachol)-induced K+ loss from a rat parotid mince were studied using 86Rb+ as a K+ marker. Both components of 86Rb+ loss were blunted by K+ and Cl- channel blockers and by removal of extracellular Ca2+, consistent with the hypothesis that 86Rb+ loss occurs via a Ca(2+)-activated K+ channel and that this cation loss serves to electrically balance a concomitant loss of the corresponding anion via one or more conductive pathways (channels). Two tissue "pools" of 86Rb+ were observed, a carbachol-sensitive pool and a carbachol-insensitive pool (approximately 70 and approximately 30% of the total 86Rb+ content, respectively). There was no evidence for a time-dependent desensitization of the muscarinic response of the carbachol-sensitive pool. Cl(-)-dependent 86Rb+ loss was not affected by HCO3- addition, suggesting that both Cl- and HCO3- secretion are accompanied by 86Rb+ loss from the same pool and thus occur from the same cells. HCO3(-)-dependent 86Rb+ loss was not enhanced by lowering the extracellular Na+ concentration, indicating that the HCO3- exit pathway is not a Na(+)-HCO3- symport. The data are consistent with the postulate that Cl- and HCO3- are secreted by rat parotid acinar cells via the same or very similar conductive transport pathways in response to muscarinic stimulation.

Suppression of carbachol-induced oscillatory Cl− secretion by forskolin in rat parotid and submandibular acinar cells

2008 ◽  
Vol 294 (3) ◽  
pp. G738-G747 ◽  
Author(s):  
Takahide Shintani ◽  
Chikara Hirono ◽  
Makoto Sugita ◽  
Yoshiko Iwasa ◽  
Yoshiki Shiba
Keyword(s):  
Membrane Potential ◽  
Salivary Glands ◽  
Acinar Cells ◽  
Fluid Secretion ◽  
Sympathetic Stimulation ◽  
Current Clamp ◽  
Inward Current ◽  
Parotid Acinar Cells ◽  
Current Clamp Mode ◽  
Rat Parotid

Sympathetic stimulation induces weak salivation compared with parasympathetic stimulation. To clarify this phenomenon in salivary glands, we investigated cAMP-induced modulation of Ca2+-activated Cl− secretion from rat parotid and submandibular acinar cells because fluid secretion from salivary glands depends on the Cl− secretion. Carbachol (Cch), a Ca2+-increasing agent, induced hyperpolarization of the cells with oscillatory depolarization in the current clamp mode of the gramicidin-perforated patch recording. In the voltage clamp mode at −80 mV, Cch induced a bumetanide-sensitive oscillatory inward current, which was larger in rat submandibular acinar cells than in parotid acinar cells. Forskolin and IBMX, cAMP-increasing agents, did not induce any marked current, but they evoked a small nonoscillatory inward current in the presence of Cch and suppressed the Cch-induced oscillatory inward current in all parotid acinar cells and half (56%) of submandibular acinar cells. In the current clamp mode, forskolin + IBMX evoked a small nonoscillatory depolarization in the presence of Cch and reduced the amplitude of Cch-induced oscillatory depolarization in both acinar cells. The oscillatory inward current estimated at the depolarized membrane potential was suppressed by forskolin + IBMX. These results indicate that cAMP suppresses Ca2+-activated oscillatory Cl− secretion of parotid and submandibular acinar cells at −80 mV and possibly at the membrane potential during Cch stimulation. The suppression may result in the weak salivation induced by sympathetic stimulation.

scholarly journals β-Adrenergic stimulation alters oligosaccharide pyrophosphoryl dolichol metabolism in rat parotid acinar cells

1985 ◽  
Vol 231 (2) ◽  
pp. 431-438 ◽  
Author(s):  
S R Grant ◽  
E E Kousvelari ◽  
D K Banerjee ◽  
B J Baum
Keyword(s):  
Half Life ◽  
Specific Radioactivity ◽  
Acinar Cells ◽  
Protein Glycosylation ◽  
Significant Enhancement ◽  
Adrenergic Stimulation ◽  
Parotid Acinar Cells ◽  
Secretory Glycoproteins ◽  
Rat Parotid ◽  
Stimulation Of

beta-Adrenergic stimulation of rat parotid acinar cells markedly increases [3H]mannose incorporation into N-linked glycoproteins [Kousvelari, Grant, Banerjee, Newby & Baum (1984) Biochem. J. 222, 17-24]. More than 90% of this protein-bound [3H]mannose was preferentially incorporated into four secretory glycoproteins. The ratio of [3H]mannose/[14C]leucine present in these individual proteins was 1.7-4-fold greater with isoproterenol-treated cells than with untreated controls. In isoproterenol-stimulated cells, [3H]mannose incorporation into mannosylphosphoryl dolichol and oligosaccharide-PP-dolichol was increased 2-3-fold over that observed in unstimulated cells. Similarly, formation of mannosylated oligosaccharide-PP-dolichol was increased approx. 4-fold in microsomes prepared from isoproterenol-treated cells. Also, turnover of oligosaccharide-PP-dolichol was significantly increased (5-fold) by β-adrenergic stimulation; the half-life for oligosaccharide-PP-dolichol decreased from 6 min in control cells to 1.2 min in isoproterenol-stimulated cells. By 15 min after isoproterenol addition to acinar cells, the specific radioactivity of parotid oligosaccharide moieties increased about 3-fold over the value observed in the absence of the agonist. Taken together, these results strongly suggest that elevation of N-linked protein glycosylation in rat parotid acinar cells after β-adrenoreceptor stimulation resulted from significant enhancement in the synthesis of mannosylphosphoryl dolichol and oligosaccharide-PP-dolichol and the turnover of oligosaccharide-PP-dolichol.

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