Regulation of ion transport in porcine gallbladder: effects of VIP and norepinephrine

1989 ◽  
Vol 257 (1) ◽  
pp. C52-C57 ◽  
Author(s):  
S. M. O'Grady ◽  
P. J. Wolters ◽  
K. Hildebrand ◽  
D. R. Brown
Keyword(s):  
Receptor Antagonist ◽  
Adrenergic Receptor ◽  
Sulfonic Acid ◽  
Short Circuit ◽  
Fold Increase ◽  
Short Circuit Current ◽  
Maximal Effect ◽  
Camp Content ◽  
Flux Measurements

The objective of this study was to investigate the effects of vasoactive intestinal peptide (VIP) and norepinephrine (NE) on Na and Cl transport across the isolated porcine gallbladder. Serosal addition of either VIP or secretin increased the short-circuit current (Isc). The half-maximal effect for VIP was 84.3 nM. The effect of VIP was mimicked by 8-bromoadenosine 3',5'-cyclic monophosphate (8-BrcAMP). Replacement of Cl with gluconate nearly abolished the effect of 8-BrcAMP on Isc, whereas HCO3 replacement with N-2-hydroxyethylpiperazine-N'-2-ethane-sulfonic acid buffer had no effect. Transepithelial flux measurements indicated that 8-BrcAMP stimulates net Cl secretion and inhibits Na absorption. Norepinephrine inhibits VIP-stimulated changes in Isc as well as the basal Isc. NE does not, however, reverse the effects of 8-BrcAMP on Isc. The effects of NE are antagonized by yohimbine (alpha 2-adrenergic receptor antagonist) but not prazosin (an alpha 1-adrenergic receptor antagonist). VIP causes a 2.5-fold increase in cAMP content in the gallbladder epithelium. This increase is blocked by NE. Serosal tetrodotoxin did not inhibit the peptide effects, indicating that VIP receptors are localized on the epithelium. Depolarization of submucosal nerves with veratrine inhibited the basal Isc and was reversible with yohimbine. This result indicated that sympathetic nerve pathways regulate Na and Cl absorption in vitro.

alpha 2-Adrenergic receptor regulation of ion transport in rabbit ileum

1982 ◽  
Vol 242 (3) ◽  
pp. G237-G242 ◽  
Author(s):  
E. B. Chang ◽  
M. Field ◽  
R. J. Miller
Keyword(s):  
Ion Transport ◽  
Adrenergic Receptor ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Maximal Effect ◽  
Rabbit Ileum ◽  
Ileal Mucosa ◽  
Receptor System ◽  
Alpha Adrenergic Receptors

Catecholamines are known to decrease short-circuit current (Isc), stimulate NaCl absorption, and inhibit HCO3 secretion in rabbit ileal mucosa in vitro. These effects appear to be mediated by alpha-adrenergic receptors because they are partially blocked by phentolamine and not by propranolol. To further characterize this receptor system, we determined the interactions of epinephrine (Epi) with alpha-subtype-selective antagonists. Prazosin (PZ), a specific alpha 1-antagonist, did not alter the Epi dose-response curve at concentrations up to 10(-5) M. Yohimbine (YO), a specific alpha 2-antagonist, completely inhibited the Epi effect on Isc. At 10(-5) M, YO increased by 70-fold the concentration of Epi required to produce a half-maximal effect (ED50; from 1.4 X 10(-7) M to 10(-5) M). YO and PZ by themselves had no significant effect on Isc in concentrations up to 10(-5) M. Clonidine, a specific alpha 2-agonist, decreased Isc with an ED50 similar to that of Epi; its effect was blocked by YO but not by PZ. Two alpha 1-selective agonists, methoxamine and phenylephrine, only caused a decrease in Isc in doses greater than 10(-5) M. This effect was reversed by YO but not by PZ. The effects of YO and PZ on Epi-modified Cl fluxes were also determined. YO completely aborted the effects of Epi on net Cl flux. No significant effects were seen with PZ. We conclude that the effects of Epi on ileal ion transport are mediated by a specific alpha 2-adrenergic receptor present in ileal mucosa and that no physiologic alpha 1-receptor function can be demonstrated.

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)

Antisecretory effect of prostaglandin D2 in rat colon in vitro: action sites

1991 ◽  
Vol 260 (6) ◽  
pp. G904-G910 ◽  
Author(s):  
K. J. Goerg ◽  
C. Diener ◽  
M. Diener ◽  
W. Rummel
Keyword(s):  
Short Circuit ◽  
Ussing Chamber ◽  
Short Circuit Current ◽  
Rat Colon ◽  
Prostaglandin D2 ◽  
Cyclic Monophosphate ◽  
Heat Stable ◽  
Antisecretory Effect ◽  
Flux Measurements

The effect of prostaglandin D2 (PGD2) on colonic ion transport was studied in the Ussing chamber. PGD2 (10(-6) M) decreased baseline short-circuit current (Isc) in two preparations of rat colon descendens, a mucosa-submucosa preparation with and a mucosa preparation without the submucosal plexus. In both preparations, PGD2 inhibited the neuronally mediated secretory responses to electric field stimulation, the sea anemone toxin ATX II, and different cholinergic agents. Unidirectional flux measurements revealed that PGD2 diminished the secretagogue-induced increase in the serosal-to-mucosal flux of Cl- and thereby inhibited net Cl- secretion. PGD2, however, had no effect on the adenosine 3',5'-cyclic monophosphate-mediated response to forskolin or vasoactive intestinal peptide or on guanosine 3',5'-cyclic monophosphate-mediated secretion induced by the heat-stable enterotoxin of Escherichia coli. The PGD2 also blocked the increase in Isc evoked by two neuronally acting inflammatory mediators, i.e., bradykinin and PGI2 in the mucosa-submucosa preparation, but had no effect on the response to PGE2. Consequently, PGD2 exerts an indirect antisecretory effect caused by an inhibition of enteric secretomotor neurons of both the submucosal and the mucosal plexus.

Receptors for vasoactive intestinal peptide and secretin on small intestinal epithelial cells

1980 ◽  
Vol 238 (3) ◽  
pp. G190-G196
Author(s):  
H. J. Binder ◽  
G. F. Lemp ◽  
J. D. Gardner
Keyword(s):  
Small Intestine ◽  
Guinea Pig ◽  
Vasoactive Intestinal Peptide ◽  
Binding Sites ◽  
Prostaglandin E1 ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Maximal Effect ◽  
Single Class

Binding of 125I-labeled vasoactive intestinal peptide (VIP) to dispersed enterocytes prepared from guinea pig small intestine was saturable, temperature dependent, and reversible, and reflected interaction of the labeled peptide with a single class of binding sites. Each enterocyte possessed approximately 60,000 binding sites and binding of the tracer to these sites could be inhibited by VIP [concentration for half-maximal effect (Kd), 12 nM] and by secretin (Kd greater than 1 micro M), but not by glucagon, gastrin, cholecystokinin, calcitonin, bombesin, litorin, physalaemin, substance P, eledoisin, serotonin, carbamylcholine, or histamine. With VIP and secretin, there was a close correlation between the relative potency for inhibition of binding of 125I-VIP and that for increasing cellular cAMP. For a given peptide, however, a 10-fold higher concentration was required for half-maximal inhibition of binding than for half-maximal stimulation of cellular cAMP. In addition to inhibiting binding of 125I-VIP and increasing cellular cAMP in enterocytes, secretin caused an increase in short-circuit current across guinea pig small intestine in vitro. Prostaglandin E1 increased cellular cAMP, but did not alter binding of 125I-VIP and the increase in cAMP caused by prostaglandin E1 plus VIP or secretin was equal to the sum of the increase caused by each agent alone.

Epithelial properties of human colonic carcinoma cell line Caco-2: effect of secretagogues

1985 ◽  
Vol 248 (5) ◽  
pp. C410-C418 ◽  
Author(s):  
E. Grasset ◽  
J. Bernabeu ◽  
M. Pinto
Keyword(s):  
Short Circuit ◽  
Short Circuit Current ◽  
Chloride Secretion ◽  
Colonic Carcinoma ◽  
Secretory Cells ◽  
Mucosal Membrane ◽  
Flux Measurements ◽  
Cell Depolarization ◽  
Epithelial Layers

Human colonic carcinoma Caco-2 cells grown in vitro form epithelial layers of highly polarized cells. Unlike colonic adsorptive cells they possess a mucosal membrane with very limited ionic conductance, even after exposure to aldosterone. When grown on filters, Caco-2 cells were sensitive to various secretagogues; these included 10(-5) M dibutyryl adenosine 3',5'-cyclic monophosphate (DBcAMP) and 10(-10) M vasoactive intestinal peptide, both of which, added serosally, enhanced the short-circuit current. The same applied to mucosal forskolin. Caco-2 cell sensitivity to serosal epinephrine was lower. Ion substitutions and 22Na-36Cl flux measurements indicated the possibility of secretagogue-dependent chloride secretion. Measurements on cells grown on Petri dishes and exposed to 1 mM DBcAMP for 1 h enabled detection of more profound modifications. Sustained 20-mV cell depolarization and a large reduction in the relative electrical resistance of the mucosal membrane were concomitant with a sizable decrease in 36Cl accumulation. These results suggest that Caco-2 cells, which to some extent resemble colonic crypt cells, possess the cAMP-dependent mucosal chloride conductance characteristic of secretory cells.

Hormonal control of chloride transport across locust rectum

1978 ◽  
Vol 56 (8) ◽  
pp. 1879-1882 ◽  
Author(s):  
J. Spring ◽  
J. Hanrahan ◽  
J. Phillips
Keyword(s):  
Chloride Transport ◽  
Short Circuit ◽  
Fold Increase ◽  
Short Circuit Current ◽  
Open Circuit ◽  
Hormonal Control ◽  
Net Flux ◽  
Hemolymph Volume ◽  
Camp Levels

Rates of ion transport across locust recta were monitored in vitro by following fluxes of 22Na+ and 36Cl−, short-circuit current (Isc), and open-circuit electropotential difference (PD) across this epithelium for several hours. Corpora cardiaca (CC) homogenates, cAMP, theophylline, and hemolymph of recently fed locusts all stimulate electrogenic transport of Cl− across locust rectum, as indicated by a two- to three-fold increase in 36Cl− net flux, Isc, and PD. Cyclic AMP caused a Cl-dependent increase in PD across the lumen-facing but not the hemocoel-facing plasma membrane of the epithelial cells. We propose that a blood-borne factor, possibly from the CC, causes an elevation in cAMP levels in rectal tissue and that this second messenger acts by increasing Cl− entry into the cell from the rectal lumen. Additional fluid absorption accompanies the resulting increase in transport of NaCl, leading to an increase in the hemolymph volume of previously dehydrated locusts.

Permeability of the rabbit colon in vitro

1977 ◽  
Vol 232 (1) ◽  
pp. F5-F9 ◽  
Author(s):  
T. Yorio ◽  
P. J. Bentley
Keyword(s):  
Amphotericin B ◽  
Short Circuit ◽  
Physiological Role ◽  
Short Circuit Current ◽  
Rabbit Colon ◽  
Flux Measurements

The rabbit colon, in vitro as everted-sac or diaphragm preparations, exhibits a transmural PD, as high as 70 mV, a short-circuit current (SCC) of 100 to 150 muA cm-3 and a resistance of 300–500 omega-cm2. It maintains these functions for at least 3 h. The SCC can be abolished by amiloride or increased by amphotericin B. Na, Cl, and K flux measurements showed a net influx (mucosa to serosa) of Na and a net efflux of K. The SCC can be accounted for by the movements of these ions. The SCC in the presence of amphotericin B was nearly equivalent to the net Na flux. Amiloride abolished the net Na transfer but did not ht (Ktrnas X 10(7)) for water was 5,000 cm s-1 while that for urea was 26 cm s-1. The rabbit provides a viable pn and the present observations appear to be consistent with its physiological role.

Potassium secretion by rabbit descending colon: effects of adrenergic stimuli

1986 ◽  
Vol 250 (4) ◽  
pp. G432-G439 ◽  
Author(s):  
P. L. Smith ◽  
R. D. McCabe
Keyword(s):  
Short Circuit ◽  
Short Circuit Current ◽  
Maximal Effect ◽  
Dependent Manner ◽  
Adrenergic Stimulation ◽  
Adrenergic Agents ◽  
Ussing Chambers ◽  
K Secretion ◽  
Potassium Secretion

Stripped rabbit distal colonic mucosa was studied in vitro in Ussing chambers to investigate the effects of adrenergic stimuli on Na+, K+, and Cl- transport. The adrenergic stimuli epinephrine and norepinephrine decrease short-circuit current in a dose-dependent manner, with a half-maximal effect at 5 X 10(-7) M and a maximal effect between 10(-5) and 10(-4) M. The effects produced by norepinephrine and epinephrine can also be elicited by the beta 1-agonist dobutamine, but not by the beta 2-agonist terbutaline or the alpha-agonist phenylephrine. In addition, the effects of adrenergic stimulation can be inhibited by the beta-antagonist propranolol but not by the muscarinic antagonist atropine, the alpha 2-antagonist yohimbine, or tetrodotoxin. The decrease in short-circuit current elicited by adrenergic stimuli is accompanied by an increase in net K+ secretion with no change in net Cl- or Na+ transport. This increase in net K+ secretion elicited by beta-adrenergic stimulation can be inhibited by trifluoperazine but not by indomethacin. These studies suggest that K+ transport by the colon can be regulated by adrenergic agents acting via beta 1-receptors.

Electrogenic Na+ absorption of rat distal colon is confined to surface epithelium: a voltage-scanning study

1993 ◽  
Vol 264 (5) ◽  
pp. C1285-C1293 ◽  
Author(s):  
A. Kockerling ◽  
D. Sorgenfrei ◽  
M. Fromm
Keyword(s):  
Short Circuit ◽  
Fold Increase ◽  
Short Circuit Current ◽  
Distal Colon ◽  
Source Model ◽  
Surface Epithelium ◽  
Electrode Position ◽  
Electrogenic Transport ◽  
Voltage Scanning

There is no quantitative assignment of large intestinal electrogenic Na+ absorption to surface epithelium and crypts so far. We determined the spatial distribution of electrogenic Na+ absorption to crypts and surface epithelium of rat late distal colon using a modified voltage-scanning technique. Voltage deflections resulting from external 30-Hz current were sensed by an extracellular microelectrode stepping at 0.7 Hz above crypt openings or surface epithelium. Local conductances were calculated applying a planar model of electrical field distribution to surface epithelium and a electrostatic disk source model to the crypts. These models were confirmed by methodological experiments where the electrode position was varied in vertical and horizontal direction. Electrogenic Na+ absorption was detected by blocking apical Na+ channels by mucosal 0.1 mM amiloride. Under control conditions surface epithelium contributed 44% (2.0 +/- 0.2 mS/cm2) and crypts 56% (2.6 +/- 0.2 mS/cm2) to the total conductance of 4.6 +/- 0.4 mS/cm2. Electrogenic Na+ absorption was induced by 6 h in vitro incubation in a medium containing 3 nM aldosterone. This caused a short-circuit current (ISC) of 12.1 +/- 0.8 mumol.h-1.cm-2, which was paralleled by a 2.5-fold increase in surface epithelial conductance to 5.1 +/- 0.4 mS/cm2, whereas crypt conductance was not significantly altered (3.0 +/- 0.2 mS/cm2). Amiloride reversed ISC to -0.8 +/- 0.1 mumol.-1.cm-2 and decreased surface epithelium conductance to 2.3 +/- 0.3 mS/cm2 but again had no significant effect on crypt conductance (2.5 +/- 0.3 mS/cm2). Sham incubation (no hormones added) for 6 h neither induced electrogenic transport nor altered local epithelial conductances.(ABSTRACT TRUNCATED AT 250 WORDS)

IL-1 stimulates intestinal myofibroblast COX gene expression and augments activation of Cl- secretion in T84 cells

1996 ◽  
Vol 271 (4) ◽  
pp. C1262-C1268 ◽  
Author(s):  
T. A. Hinterleitner ◽  
J. I. Saada ◽  
H. M. Berschneider ◽  
D. W. Powell ◽  
J. D. Valentich
Keyword(s):  
Gene Expression ◽  
Short Circuit ◽  
Interleukin 1 ◽  
Fold Increase ◽  
Short Circuit Current ◽  
Mrna Levels ◽  
T84 Cells ◽  
Important Mediator ◽  
Cox 1

Because interleukin-1 (IL-1) is an important mediator in the inflamed intestine, its effects on enterocyte-subepithelial myofibroblast (SEMF) interaction were investigated in vitro. Acutely juxtaposing T84 cells with 18Co or P2JF SEMF preincubated with IL-1 alpha significantly enhanced T84 short-circuit current (Isc) responsiveness to secretagogues in comparison to SEMF not activated by IL-1 alpha. The sensitivity of T84 cell Isc to Ca(2+)-dependent, but not adenosine 3',5'-cyclic monophosphate-dependent, secretagogues was augmented by IL-1 alpha-treated SEMF. These effects of IL-1 alpha are directly correlated with SEMF prostaglandin E2 (PGE2) production. Both IL-1 alpha augmentation of Cl secretagogue responsiveness and PGE2 formation were inhibited by IL-1 receptor antagonist. Within 5 h, IL-1 alpha stimulated a 10-fold increase in cyclooxygenase (COX)-2 steady-state mRNA levels in 18Co cells. In contrast, COX-1 message levels increased more slowly to two- to threefold above control levels after 24 h incubation. These results demonstrate that the proinflammatory cytokine IL-1 alpha accentuates intestinal SEMF augmentation of enterocyte responsiveness to Ca(2+)-dependent CI-secretagogues. PGE2 is an important mediator of SEMF-enterocyte interaction. The effects of IL-1 alpha on SEMF PGE2 productions are, at least in part, due to stimulation of COX gene expression.

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