Endothelin stimulates chloride secretion across canine tracheal epithelium

1991 ◽  
Vol 261 (2) ◽  
pp. L188-L194 ◽  
Author(s):  
P. I. Plews ◽  
Z. A. Abdel-Malek ◽  
C. A. Doupnik ◽  
G. D. Leikauf
Keyword(s):  
Epithelial Cells ◽  
Short Circuit ◽  
Second Messengers ◽  
Short Circuit Current ◽  
Chloride Secretion ◽  
Tracheal Epithelium ◽  
Membrane Phospholipids ◽  
Cl Secretion ◽  
Tracheal Epithelial Cells ◽  
Tracheal Epithelial

The endothelins (ET) are a group of isopeptides produced by a number of cells, including canine tracheal epithelial cells. Because these compounds are endogenous peptides that may activate eicosanoid metabolism, we investigated the effects of ET on Cl secretion in canine tracheal epithelium. Endothelin 1 (ET-1) was found to produce a dose-dependent change in short-circuit current (Isc) that increased slowly and reached a maximal value within 10-15 min. When isopeptides of ET were compared, 300 nM ET-1 and ET-2 produced comparable maximal increases in Isc, whereas ET-3 produced smaller changes in Isc (half-maximal concentrations of 2.2, 7.2, and 10.4 nM, respectively). Ionic substitution of Cl with nontransported anions, iodide and gluconate, reduced ET-1-induced changes in Isc. Furthermore, the response was inhibited by the NaCl cotransport inhibitor, furosemide. In paired tissues, ET-1 significantly increased mucosal net 36Cl flux without significant effect on 22Na flux. The increase in Isc induced by ET was diminished by pretreatment with indomethacin. The second messengers mediating the increase in Isc were investigated in cultured canine tracheal epithelial cells. ET-1 stimulated the release of [3H]arachidonate from membrane phospholipids, increased intracellular Ca2+ (occasionally producing oscillations), and increased adenosine 3',5'-cyclic monophosphate accumulation. The latter was diminished by indomethacin. Thus ET is a potent agonist of Cl secretion (with the isopeptides having the following potency: ET-1 greater than or equal to ET-2 greater than ET-3) and acts, in part, through a cyclooxygenase-dependent mechanism.

Changes in intracellular sodium with chloride secretion in dog tracheal epithelium

1986 ◽  
Vol 250 (4) ◽  
pp. C646-C650 ◽  
Author(s):  
S. R. Shorofsky ◽  
M. Field ◽  
H. A. Fozzard
Keyword(s):  
Steady State ◽  
Kinetic Models ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Chloride Secretion ◽  
Tracheal Epithelium ◽  
Cl Secretion ◽  
Intracellular Na Activity ◽  
Sufficient Energy ◽  
Stimulation Of

Na-selective microelectrodes were employed to investigate the mechanism of Cl secretion by canine tracheal epithelium. In control tissues with a mean short-circuit current (Isc) of 30.1 microA/cm2, the intracellular Na activity (aiNa) was 10.7 mM. Following steady-state stimulation of Cl secretion with epinephrine (Isc = 126.4 microA/cm2), aiNa was 21.3 mM. These data indicate that there is sufficient energy in the Na gradient to drive Cl secretion by this tissue. When analyzed with simple kinetic models for the Na-K pump, they also suggest that the basolateral entry step involves the Na-K-2Cl cotransporter.

Phorbol ester and okadaic acid regulation of Na-2Cl-K cotransport in rabbit tracheal epithelial cells

1996 ◽  
Vol 271 (1) ◽  
pp. C338-C346 ◽  
Author(s):  
C. M. Liedtke ◽  
L. Thomas
Keyword(s):  
Epithelial Cells ◽  
Okadaic Acid ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Term Treatment ◽  
Secretory Cells ◽  
Adrenergic Stimulation ◽  
Short Term Treatment ◽  
Tracheal Epithelial Cells ◽  
Tracheal Epithelial

We evaluated a role for protein kinase C (PKC) in the regulation of rabbit tracheal epithelial Na-Cl(K) cotransport. Short-term treatment with phorbol 12-myristate 13-acetate (PMA) dose dependently increased bumetanide-sensitive Na and Cl efflux and elevated staurosporine- and bumetanide-sensitive Na, Cl, and K uptake. PMA and the alpha 2A-adrenergic agonist guanabenz both induced contransport with a stoichiometry of 2 Cl:1 Na and 2 Cl:1 Rb and elevated staurosporine-sensitive PKC activity in cytosolic and particulate fractions. Prolonged PMA treatment did not sustain bumetanide-sensitive 2 Cl:1 Na and 2 Cl:1 Rb transport but did block stimulation of bumetanide-sensitive transport by PMA or guanabenz and elevation of PKC activity by PMA and guanabenz in a particulate fraction. Cells treated with okadaic acid exhibited a staurosporine- and bumetanide-sensitive 2 Cl:1 Na and 2 Cl:1 Rb uptake. In cultured monolayers, basolateral perfusion with epinephrine, isoproterenol, or PMA increased short-circuit current (Isc). Basolateral application of bumetanide reduced elevated Isc to baseline levels, indicating a role for Cl secretory cells in a reconstituted tracheal epithelium. Pretreatment of transmonolayer cultures with PMA diminished the stimulatory response to epinephrine. These results indicate that, in rabbit tracheal epithelial cells, alpha-adrenergic stimulation activated Na-2Cl-K cotransport and that PKC is a critical effector in this process.

Regulation of chloride secretion in dog tracheal epithelium by protein kinase C

1987 ◽  
Vol 253 (6) ◽  
pp. C802-C808 ◽  
Author(s):  
R. A. Barthelson ◽  
D. B. Jacoby ◽  
J. H. Widdicombe
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Chloride Secretion ◽  
Tracheal Epithelium ◽  
Base Line ◽  
Cl Secretion ◽  
Cell Extracts ◽  
Kinase C

The effects of stimulating protein kinase C on Cl- secretion across dog tracheal epithelium were investigated. The phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA), and the synthetic diacylglycerol, 1-oleolyl-2-acetylglycerol (OAG), which stimulate protein kinase C (PKC), both stimulated short-circuit current (Isc) with Kd of 10 nM and 1 microM, respectively. In Cl(-)-free solution, the increases in Isc were virtually abolished, suggesting that these compounds stimulate Cl- secretion, a hypothesis confirmed for TPA by measurement of 36Cl- fluxes. The stimulations of Cl- secretion were not sensitive to indomethacin, nor were cAMP levels elevated during stimulation. In addition to its transient stimulatory effect, TPA at high doses caused the eventual lowering of the base-line Isc and a block of subsequent stimulation by cAMP-mediated agonists. This was probably not the result of toxicity or an effect on adenylate cyclase or on cAMP-dependent protein kinase. Cell extracts from both cultured and native dog tracheal epithelial cells showed strong PKC activities. These results suggest that PKC may play a role in regulating Cl- secretion across dog tracheal epithelium.

Effect of vasoactive intestinal peptide on ion transport across dog tracheal epithelium

1983 ◽  
Vol 55 (6) ◽  
pp. 1844-1848 ◽  
Author(s):  
I. Nathanson ◽  
J. H. Widdicombe ◽  
P. J. Barnes
Keyword(s):  
Epithelial Cells ◽  
Vasoactive Intestinal Peptide ◽  
Short Circuit ◽  
Statistical Significance ◽  
Short Circuit Current ◽  
Open Circuit ◽  
Tracheal Epithelium ◽  
Cl Secretion ◽  
High Affinity ◽  
Stimulation Of

Under short-circuit conditions, vasoactive intestinal peptide (VIP) did not alter net Na+ movement but selectively stimulated net Cl- secretion across dog tracheal epithelium with a high affinity (Km congruent to 10(-8) M). The increase in Cl- secretion was not different from the rise in short-circuit current (Isc). However, stimulation of Cl- secretion was not maximal, because the addition of isoproterenol (10(-6) M) to VIP-treated tissues further increased the Isc by 54%. The effect of exogenous VIP was not blocked by a combination of atropine, phentolamine, propranolol (10(-5) or 10(-6) M), or tetrodotoxin (10(-6) M). Under open-circuit conditions, VIP caused an increase in the net secretion of Cl- and Na+, but the changes did not reach statistical significance. We conclude that VIP acts directly on receptors on the surface of epithelial cells to stimulate active Cl- secretion. The abundance of VIP nerves in the submucosa suggests that VIP may be important in regulation of fluid movement across the epithelium.

Atriopeptin stimulates chloride secretion in cultured shark rectal gland cells

1991 ◽  
Vol 260 (5) ◽  
pp. C1125-C1130 ◽  
Author(s):  
K. J. Karnaky ◽  
J. D. Valentich ◽  
M. G. Currie ◽  
W. F. Oehlenschlager ◽  
M. P. Kennedy
Keyword(s):  
Epithelial Cells ◽  
Plasma Membranes ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Chloride Secretion ◽  
Rectal Gland ◽  
Cl Secretion ◽  
Intracellular Cgmp ◽  
Similar Dose ◽  
Shark Rectal Gland

Monolayer cultures of shark rectal gland (SRG) epithelial cells were treated with atriopeptin (AP), and the effects on Cl- secretion and intracellular guanosine 3',5'-cyclic monophosphate (cGMP) accumulation were examined. Basolateral or apical exposure to 10(-7) M AP markedly stimulated (8-fold) Cl(-)-dependent, bumetanide-sensitive, short-circuit current (Isc). The AP-stimulated Isc exhibited transient oscillations before reaching a steady state. This behavior is not observed when Isc is activated by other secretagogues such as vasoactive intestinal peptide, 2-chloroadenosine, forskolin, or ionomycin. Intracellular cGMP was concomitantly elevated (10-fold) by 10(-7) M AP. Both Isc stimulation and cGMP accumulation responses exhibited a similar dose dependency beginning at an AP concentration of 1 nM. The bilateral response to AP suggests the presence of receptors on both apical and basolateral plasma membranes. These results are the first demonstration of a direct effect of AP on Cl(-)-secreting epithelial cells. These data also suggest a role for cGMP in mediating Cl- secretion in these cells.

HOCl effects on tracheal epithelium: conductance and permeability measurements

1995 ◽  
Vol 78 (4) ◽  
pp. 1330-1338 ◽  
Author(s):  
Y. Guo ◽  
L. A. Schneider ◽  
O. D. Wangensteen
Keyword(s):  
Epithelial Cells ◽  
Hypochlorous Acid ◽  
Short Circuit ◽  
Electrical Conductance ◽  
Short Circuit Current ◽  
Tracheal Epithelium ◽  
P Values ◽  
Solute Permeability ◽  
Age Dependent ◽  
Tracheal Epithelial

It is speculated that hypochlorous acid (HOCl), produced by neutrophils, can disrupt the tracheal epithelial barrier without damage to epithelial cells. This was investigated with solute permeability (P) and electrical conductance (G) measurements on tracheae from 4-wk-old rabbits. A new system for epithelial bioelectric measurements on intact tracheae was developed and validated. Control values of G, short-circuit current, and spontaneous potential difference were 4.9 +/- 0.5 (SE) mS/cm2, 42.6 +/- 4.7 microA/cm2, and 8.9 +/- 1.0 mV (lumen negative), respectively (n = 5). Control P values for sucrose, inulin, and Dextran-20 were 5.14 +/- 0.48, 0.63 +/- 0.10, and 0.057 +/- 0.007 x 10(-7) cm/s, respectively (n = 6). Tracheae treated with HOCl had no effect; 6 mM HOCl, a concentration that could exist in infected airways, significantly increased both P and G (about two- to fourfold) without damage to epithelial cells; and 12 and 30 mM HOCl caused more than 10-fold increases for both P and G with cell disruption. Vitamin C blocked epithelial damage caused by 30 mM HOCl. Tracheae from 1-wk-old rabbits were significantly more sensitive to HOCl than those from 4-wk-old or adult rabbits. This study validated a new bioelectric measurement system and showed that HOCl has both dose- and age-dependent effects on the tracheal epithelium.

Electrical properties of dog tracheal epithelial cells grown in monolayer culture

1984 ◽  
Vol 246 (3) ◽  
pp. C355-C359 ◽  
Author(s):  
D. L. Coleman ◽  
I. K. Tuet ◽  
J. H. Widdicombe
Keyword(s):  
Electrical Properties ◽  
Epithelial Cells ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Airway Epithelia ◽  
Tracheal Epithelial Cells ◽  
Monolayer Cultures ◽  
Tracheal Epithelial ◽  
Pg E2 ◽  
Water And Ion Transport

Epithelial cells from dog trachea, when grown in tissue culture, formed confluent monolayers 5-6 days after plating. After 8-10 days, domes [mean diam 356 +/- (SE) 53 micron] appeared in monolayers grown in collagen-coated flasks. When grown on polycarbonate filters coated with collagen, a detectable resistance (greater than 5 omega X cm2) and transepithelial potential difference (PD) (greater than 0.1 mV) developed 6 days after plating and increased to approximately 15 omega X cm2 and 15 mV at 10 days. Serosal ouabain (10(-4) M) abolished PD and short-circuit current (Isc). Luminal ouabain had no effect. Luminal amiloride (10(-4) M) and serosal bumetanide (10(-4) M) each decreased PD and Isc. However, a combination of both of these drugs did not abolish Isc. Isoproterenol (10(-5) M), dibutyryl adenosine 3',5'-cyclic monophosphate (10(-3) M), vasoactive intestinal peptide (10(-7) M), prostaglandin (PG) E2 (10(-5) M), PGF2 alpha (10(-5) M), and bradykinin (10(-5) M) each increased PD and Isc. Thus these monolayer cultures maintain electrical properties resembling those of the original tissue. This preparation may prove useful for the study of water and ion transport by airway epithelia.

Mechanisms of sodium and chloride transport across equine tracheal epithelium

1990 ◽  
Vol 259 (6) ◽  
pp. L459-L467 ◽  
Author(s):  
G. J. Tessier ◽  
T. R. Traynor ◽  
M. S. Kannan ◽  
S. M. O3'Grady
Keyword(s):  
Chloride Transport ◽  
Short Circuit ◽  
Basolateral Membrane ◽  
Short Circuit Current ◽  
Ringer Solution ◽  
Tracheal Epithelium ◽  
Ussing Chambers ◽  
Cl Secretion ◽  
Cotransport System ◽  
Ethanesulfonic Acid

Equine tracheal epithelium, stripped of serosal muscle, mounted in Ussing chambers, and bathed in plasmalike Ringer solution generates a serosa-positive transepithelial potential of 10–22 mV and a short-circuit current (Isc) of 70–200 microA/cm2. Mucosal amiloride (10 microM) causes a 40–60% decrease in Isc and inhibits the net transepithelial Na flux by 95%. Substitution of Cl with gluconate resulted in a 30% decrease in basal Isc. Bicarbonate substitution with 20 mM N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid decreased the Isc by 21%. The Cl-dependent Isc was inhibited by serosal addition of 1 mM amiloride. Bicarbonate replacement or serosal amiloride (1 mM) inhibits the net Cl flux by 72 and 69%, respectively. Bicarbonate replacement significantly reduces the effects of serosal amiloride (1 mM) on Isc, indicating its effect is HCO3 dependent. Addition of 8-bromoadenosine 3',5'-cyclic monophosphate (8-BrcAMP; 100 microM) causes a 40% increase in Isc. This effect is inhibited by subsequent addition of 10 microM serosal bumetanide. Bumetanide (10 microM) reduces net Cl secretion following stimulation with 8-BrcAMP (100 microM). Serosal addition of BaCl2 (1 mM) causes a reduction in Isc equal to that following Cl replacement in the presence or absence of 100 microM cAMP. These results suggest that 1) Na absorption depends on amiloride-inhibitable Na channels in the apical membrane, 2) Cl influx across the basolateral membrane occurs by both a Na-H/Cl-HCO3 parallel exchange mechanism under basal conditions and by a bumetanide-sensitive Na-(K?)-Cl cotransport system under cAMP-stimulated conditions, and 3) basal and cAMP-stimulated Cl secretion depends on Ba-sensitive K channels in the basolateral membrane.

Role of calcium in chloride secretion mediated by cAMP pathway activation in rabbit distal colon mucosa

1993 ◽  
Vol 264 (2) ◽  
pp. G252-G260 ◽  
Author(s):  
V. Calderaro ◽  
E. Chiosi ◽  
R. Greco ◽  
A. M. Spina ◽  
A. Giovane ◽  
...  
Keyword(s):  
Short Circuit ◽  
Fold Increase ◽  
Short Circuit Current ◽  
Distal Colon ◽  
Chloride Secretion ◽  
Free Medium ◽  
Cl Secretion ◽  
Pathway Activation ◽  
Flux Measurements ◽  
Pg E2

Effects of Ca2+ on adenosine 3',5'-cyclic monophosphate (cAMP)-mediated Cl- secretion were investigated in intact mucosa and isolated crypt cells of rabbit descending colon. Addition of 10 microM prostaglandin (PG)E2 or forskolin to tissues incubated in Ca(2+)-free medium increased the size of short-circuit current (Isc) and Cl- secretion as estimated by unidirectional 36Cl flux measurements (net flux = -2.31 +/- 0.24 vs. -1.22 +/- 0.10 mueq.h-1.cm-2, n = 4, P < 0.001). Addition of 10 microM PGE2 to tissues incubated in 1.2 mM Ca2+ Ringer induced a 7-fold increase in mean cAMP level, whereas it produced an 11-fold increase in tissues exposed to Ca(2+)-free medium. Membrane preparations from whole mucosa incubated in Ca(2+)-free medium displayed a cyclic nucleotide phosphodiesterase activity significantly lower than controls (18.76 +/- 0.54 vs. 31.20 +/- 0.39 pmol cAMP. mg protein-1.min-1, means +/- SE, n = 4, P < 0.001). Ca2+ removal also affected adenylate cyclase (AC) responsiveness to agonists; AC activity increased in controls by 54 and 226% after stimulation with 10 microM PGE2 and forskolin, respectively, but it increased more (77 and 325%, respectively) after incubation in Ca(2+)-free solutions.(ABSTRACT TRUNCATED AT 250 WORDS)

Sign in / Sign up

Export Citation Format

Share Document