scholarly journals Role of the BK channel (KCa1.1) during activation of electrogenic K+ secretion in guinea pig distal colon

2012 ◽  
Vol 303 (12) ◽  
pp. G1322-G1334 ◽  
Author(s):  
Jin Zhang ◽  
Susan T. Halm ◽  
Dan R. Halm
Keyword(s):  
Guinea Pig ◽  
Apical Membrane ◽  
Splice Variants ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Distal Colon ◽  
Bk Channel ◽  
Prostaglandin E ◽  
Maximal Inhibition ◽  
K Secretion

Secretagogues acting at a variety of receptor types activate electrogenic K+ secretion in guinea pig distal colon, often accompanied by Cl− secretion. Distinct blockers of KCa1.1 (BK, Kcnma1), iberiotoxin (IbTx), and paxilline inhibited the negative short-circuit current ( Isc) associated with K+ secretion. Mucosal addition of IbTx inhibited epinephrine-activated Isc (epi Isc) and transepithelial conductance (epi Gt) consistent with K+ secretion occurring via apical membrane KCa1.1. The concentration dependence of IbTx inhibition of epi Isc yielded an IC50 of 193 nM, with a maximal inhibition of 51%. Similarly, IbTx inhibited epi Gt with an IC50 of 220 nM and maximal inhibition of 48%. Mucosally added paxilline (10 μM) inhibited epi Isc and epi Gt by ∼50%. IbTx and paxilline also inhibited Isc activated by mucosal ATP, supporting apical KCa1.1 as a requirement for this K+ secretagogue. Responses to IbTx and paxilline indicated that a component of K+ secretion occurred during activation of Cl− secretion by prostaglandin-E2 and cholinergic stimulation. Analysis of KCa1.1α mRNA expression in distal colonic epithelial cells indicated the presence of the ZERO splice variant and three splice variants for the COOH terminus. The presence of the regulatory β-subunits KCaβ1 and KCaβ4 also was demonstrated. Immunolocalization supported the presence of KCa1.1α in apical and basolateral membranes of surface and crypt cells. Together these results support a cellular mechanism for electrogenic K+ secretion involving apical membrane KCa1.1 during activation by several secretagogue types, but the observed K+ secretion likely required the activity of additional K+ channel types in the apical membrane.

Aldosterone stimulates K secretion prior to onset of Na absorption in guinea pig distal colon

1994 ◽  
Vol 266 (2) ◽  
pp. C552-C558 ◽  
Author(s):  
D. R. Halm ◽  
S. T. Halm
Keyword(s):  
Guinea Pig ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Distal Colon ◽  
Ussing Chambers ◽  
High Affinity Receptor ◽  
Additive Increase ◽  
K Secretion ◽  
Affinity Receptor

Distal colon from guinea pig was stimulated in vitro by aldosterone in Ussing chambers that allowed measurement of short-circuit current (Isc) and tissue conductance (Gt). The response to aldosterone was delayed by approximately 20 min and resulted in a negative Isc, consistent with K secretion. Approximately 1 h later the Isc began to increase and eventually became positive, consistent with subsequent stimulation of Na absorption. The Na-absorptive response could be inhibited by mucosal amiloride without altering the rate of K secretion. Similarly, K secretion could be inhibited by serosal bumetanide without altering Na absorption. In the presence of spironolactone, actinomycin D, or cycloheximide, aldosterone failed to stimulate both K secretion and Na absorption. A dose response to aldosterone provided an apparent Kd of 2.6 +/- 0.5 nM, consistent with a high-affinity receptor coupled to this secretory response. Stimulation by the K secretagogue epinephrine did not produce an additive increase in K secretion, suggesting that the same cell type responds to both aldosterone and epinephrine and that the protein induced by aldosterone was not one of the membrane proteins responsible for K secretion.

Active K transport across rabbit distal colon: relation to Na absorption and Cl secretion

1986 ◽  
Vol 251 (2) ◽  
pp. C252-C267 ◽  
Author(s):  
D. R. Halm ◽  
R. A. Frizzell
Keyword(s):  
Apical Membrane ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Distal Colon ◽  
Secretory Process ◽  
Cl Secretion ◽  
K Secretion ◽  
Transport Inhibitors ◽  
Unidirectional Fluxes ◽  
K Transport

We measured isotopic unidirectional fluxes of K to elucidate the mechanisms of active K transport across the distal colon of the rabbit. Separate pathways for active K absorption and active K secretion were detected using various transport inhibitors and stimulators. The rate and direction of net K transport depend on the activities of these two pathways. K absorption was reduced by orthovanadate (both solutions) or serosal Ba, consistent with ATPase-dependent uptake of K across the apical membrane and exit via a Ba-sensitive basolateral K conductance. K secretion was inhibited by serosal ouabain or mucosal Ba, indicating that K secretion involves basolateral uptake via the Na-K pump and apical exit via a Ba-sensitive K conductance. Active K secretion appears to be electrogenic, since inhibition by ouabain produced equivalent changes in the net K flux and short-circuit current. Addition of bumetanide to the serosal solution or the removal of either Na or Cl from the serosal solution inhibited K secretion; mucosal solution amiloride was without effect. These results indicate that this K secretory process is independent of electrogenic Na absorption but is mechanistically similar to Cl secretory processes. Both epinephrine and prostaglandin E2 (PGE2) stimulate K secretion, but only PGE2 also stimulates Cl secretion. The response to these secretogogues suggests that the mechanisms underlying K and Cl secretion are closely linked but can be regulated independently.

scholarly journals Prostanoids stimulate K secretion and Cl secretion in guinea pig distal colon via distinct pathways

2001 ◽  
Vol 281 (4) ◽  
pp. G984-G996 ◽  
Author(s):  
Dan R. Halm ◽  
Susan Troutman Halm
Keyword(s):  
Guinea Pig ◽  
Rank Order ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Distal Colon ◽  
Prostanoid Receptors ◽  
Cl Secretion ◽  
Inflammatory Conditions ◽  
K Secretion

Short-circuit current ( I sc) and transepithelial conductance ( G t) were measured in guinea pig distal colonic mucosa isolated from submucosa and underlying muscle layers. Indomethacin (2 μM) and NS-398 (2 μM) were added to suppress endogenous production of prostanoids. Serosal addition of PGE2 (10 nM) stimulated negative I scconsistent with K secretion, and concentrations >30 nM stimulated positive I sc consistent with Cl secretion. PGE2 also stimulated G t at low and high concentrations. Dose responses to prostanoids specific for EP prostanoid receptors were consistent with stimulating K secretion through EP2 receptors, based on a rank order potency (from EC50 values) of PGE2 (1.9 nM) > 11-deoxy-PGE1 (8.3 nM) > 19( R)-hydroxy-PGE2 (13.9 nM) > butaprost (67 nM) > 17-phenyl-trinor-PGE2 (307 nM) ≫ sulprostone (>10 μM). An isoprostane, 8-iso-PGE2, stimulated K secretion with an EC50 of 33 nM. Cl secretory response was stimulated by PGD2 and BW-245C, a DP prostanoid receptor-specific agonist: BW-245C (15 nM) > PGD2 (30 nM) > PGE2 (203 nM). Agonists specific for FP, IP, and TP prostanoid receptors were ineffective in stimulating I sc and G t at concentrations <1 μM. These results indicate that PGE2stimulated electrogenic K secretion through activation of EP2 receptors and electrogenic KCl secretion through activation of DP receptors. Thus stimulation of Cl secretion in vivo would occur either via physiological concentrations of PGD2(<100 nM) or pathophysiological concentrations of PGE2(>100 nM) that could occur during inflammatory conditions.

Apical nonspecific cation conductances in rabbit cecum

1994 ◽  
Vol 266 (3) ◽  
pp. G475-G484 ◽  
Author(s):  
J. H. Sellin ◽  
W. P. Dubinsky
Keyword(s):  
Epithelial Transport ◽  
Apical Membrane ◽  
Divalent Cations ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Distal Colon ◽  
Na Channel ◽  
Similar Response ◽  
Electrogenic Transport

Rabbit cecum exhibits electrogenic Na absorption in vitro. However, because this transport process is not inhibited by amiloride nor does it demonstrate saturation kinetics typical of the amiloride-inhibitable Na channel, we considered whether the cecal transporter represented one of a recently described family of nonselective cation conductances or channels (NSCC). Both transepithelial and vesicle studies demonstrated that K, Cs, and Rb were transported via an apical conductance. Electrogenic transport was inhibited by divalent cations including Ca, Mg, and Ba but was unaffected by either lanthanum or gadolinium. Parallel studies in distal colon did not exhibit a similar response to either K substitution or Ba inhibition. Phenamil, verapamil, and nicardipine significantly inhibited the short-circuit current (Isc). stimulated by nominal Ca- and Mg-free conditions. Flux studies demonstrated a correlation between changes in Isc and Na transport. Microelectrode impalement studies suggested that there may be both NSCC and K conductance in the apical membrane. Planar bilayer studies identified a 190-pS cation channel that may correlate with the macroscopic transport properties of this epithelium. These studies are consistent with a model of cecal Na absorption mediated by a NSCC in the apical membrane; this may be the mechanism underlying the distinct epithelial transport characteristics of this intestinal segment.

5-HT activates nitric oxide-generating neurons to stimulate chloride secretion in guinea pig distal colon

1998 ◽  
Vol 275 (4) ◽  
pp. G829-G834 ◽  
Author(s):  
Atsukazu Kuwahara ◽  
Hirofumi Kuramoto ◽  
Makoto Kadowaki
Keyword(s):  
Nitric Oxide ◽  
Guinea Pig ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Distal Colon ◽  
Chloride Secretion ◽  
Evoked Response ◽  
Evoked Responses ◽  
No Donor ◽  
Simultaneous Application

The participation of nitric oxide (NO) in serotonin (5-hydroxytryptamine; 5-HT)-evoked chloride secretion in guinea pig distal colon was examined. Submucosal/mucosal segments were mounted in Ussing flux chambers, and an increase in short-circuit current ( I sc) was used as an index of secretion. Addition of 5-HT to the serosal side produced a concentration-dependent (10−7–10−5M) increase in I sc caused by chloride secretion. N G-nitro-l-arginine (l-NNA) significantly reduced the 5-HT-evoked early (P-1) and late (P-2) responses to 61.1 and 70.6% of control, respectively. Neurally evoked response was also inhibited by l-NNA. The NO donor sodium nitroprusside (SNP, 10−4 M) increased basal I sc mainly because of chloride secretion. The SNP-evoked response was significantly reduced by tetrodotoxin but was unchanged by atropine or indomethacin. These results suggest that the 5-HT-evoked increase in I sc is associated with an NO-generating mechanism. Atropine significantly reduced the 5-HT (10−5 M)-evoked P-1 and P-2 responses to 71.8 and 19.7% of control, respectively. Simultaneous application of atropine andl-NNA further decreased the 5-HT-evoked responses more than either drug alone; application ofl-NNA and atropine decreased the 5-HT-evoked P-1 and P-2 responses to 68.5 and 39.2% of atropine-treated tissues, respectively. These results suggest that noncholinergic components of P-1 and P-2 responses are 71.8 and 19.7% of control, respectively, and that NO components of P-1 and P-2 responses are 32 and 61%, respectively, of the noncholinergic component of the 5-HT-evoked responses. The results provide evidence that NO may participate as a noncholinergic mediator of 5-HT-evoked chloride secretion in guinea pig distal colon.

Apical P2Y4 purinergic receptor controls K+ secretion by vestibular dark cell epithelium

2001 ◽  
Vol 281 (1) ◽  
pp. C282-C289 ◽  
Author(s):  
Daniel C. Marcus ◽  
Margaret A. Scofield
Keyword(s):  
Apical Membrane ◽  
Purinergic Receptor ◽  
Short Circuit ◽  
Short Circuit Current ◽  
G Protein Coupled Receptors ◽  
Disulfonic Acid ◽  
Dark Cell ◽  
K Secretion ◽  
G Protein Coupled ◽  
Uridine Nucleotides

It was previously shown that K+ secretion by vestibular dark cell epithelium is under control of G protein-coupled receptors of the P2Y family in the apical membrane that are activated by both purine and uridine nucleotides (P2Y2, P2Y4, or P2Y6). The present study was conducted to determine the subtype of purinergic receptor and to test whether these receptors undergo desensitization. The transepithelial short-circuit current represents electrogenic K+ secretion and was found to be reduced by UTP, ATP, and diadenosine tetraphosphate, but not UDP. Neither pyridoxalphosphate-6-azophenyl-2′,4′-disulfonic acid (PPADS, 30 μM) nor suramin (100 μM) inhibited the effect of UTP. The potencies of the agonists were consistent with rodent P2Y4and P2Y2, but not P2Y6, receptors. The ineffectiveness of suramin was consistent with P2Y4, but not P2Y2. Transcripts for both P2Y2 and P2Y4 were found in vestibular labyrinth. Sustained exposure to ATP or UTP for 15 min caused a constant depression of short-circuit current with no apparent desensitization. The results support the conclusion that regulation of K+ secretion across vestibular dark cell epithelium occurs by P2Y4 receptors without desensitization of the response.

Extracellular ATP stimulates K+ secretion across cultured human airway epithelium

1997 ◽  
Vol 272 (6) ◽  
pp. L1084-L1091 ◽  
Author(s):  
L. L. Clarke ◽  
T. Chinet ◽  
R. C. Boucher
Keyword(s):  
Airway Epithelium ◽  
Apical Membrane ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Ringer Solution ◽  
Extracellular Atp ◽  
Cl Secretion ◽  
Human Airway ◽  
Human Airway Epithelium ◽  
K Secretion

Extracellular ATP applied to the luminal side of human airway epithelium (HAE) activates an apical membrane Cl- conductance and transepithelial Cl- secretion. However, in some HAE preparations, we have found that luminal ATP induces a change in short-circuit current (Isc), consistent with K+ secretion. Using intracellular microelectrodes and radioisotopic flux studies, we investigated whether extracellular ATP regulates transepithelial K+ secretion in primary HAE cultures. In physiological Ringer solution, HAE had a negligible electrochemical driving force for Cl- secretion (DFCl), and luminal ATP induced a change in Isc opposite in polarity to Cl- secretion. Intracellular microelectrode measurements indicated that the "reversed" Isc was associated with activation of a hyperpolarizing (K+) conductance in the apical membrane. Radioisotope studies of HAE pretreated with amiloride to induce a favorable DFCl revealed that luminal ATP stimulates a small 42K secretory flux concurrently with Cl- secretion. In ion-substituted Ringer solution, luminal ATP stimulated both the outward (K+) current and the inward (Cl-) current with approximately equal potency (approximately 10(-6) M). We conclude that luminal ATP activates an apical membrane K+ conductance and transepithelial K+ secretion across HAE.

scholarly journals Regulation of electrolyte transport with IL-1β in rabbit distal colon

1995 ◽  
Vol 4 (1) ◽  
pp. 61-66 ◽  
Author(s):  
F. R. Homaidan ◽  
H. Desai ◽  
L. Zhao ◽  
G. Broutman ◽  
R. Burakoff
Keyword(s):  
Inflammatory Bowel Disease ◽  
Protein Synthesis ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Distal Colon ◽  
Chloride Secretion ◽  
Electrolyte Transport ◽  
Interleukin 1Β ◽  
Prostaglandin E ◽  
Inflammatory Bowel

Interletrkin-1β levels are elevated in inflammatory bowel disease. In this study the mechanism by which interleukin-1β affects electrolyte transport in the rabbit distal colon, was investigated. Interleukin-1β caused a delayed increase in short-circuit current (Isc) which was attributed to protein synthesis since the effect was inhibited by cycloheximide. The interleukin-1β induced increase in Iscwas not affected by amiloride treatment but was completely inhibited by bumetanide or in chloride-free buffer and by indomethacin. Prostaglandin E2levels increased in tissue treated with interleukin-1β, but this increase was reversed by cycloheximide. These data suggest that interleukin-1β causes its effect via a yet to be identified second messenger, by increasing chloride secretion through a prostaglandin E2mediated mechanism.

H2 receptors mediate cyclical chloride secretion in guinea pig distal colon

1990 ◽  
Vol 258 (6) ◽  
pp. G887-G893 ◽  
Author(s):  
Y. Z. Wang ◽  
H. J. Cooke
Keyword(s):  
Guinea Pig ◽  
Chloride Transport ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Distal Colon ◽  
Chloride Secretion ◽  
Chloride Ions ◽  
H2 Antagonist ◽  
H2 Receptors ◽  
Peak Increase

We tested the hypothesis that histamine mediates ion secretion in the guinea pig distal colon by stimulating H2 receptors on submucosal neurons. Serosal addition of histamine evoked a transient increase in short-circuit current (Isc) followed by recurrent cyclical increases in Isc. The transient phase of the response was examined previously and was not investigated in these studies. Histamine (1.5-2.5 x 10(-5) M) evoked a peak increase in Isc of 177 +/- 25 microA/cm2 at intervals of 5 min for 1-2 h. The duration of each recurrent cycle averaged 2.1 +/- 0.3 min. The H2 agonist dimaprit evoked recurrent cycles that had larger amplitudes than those caused by histamine. In the presence of histamine or dimaprit, the amplitude of the first cycle of the response was always less than subsequent cycles, regardless of the initial concentration. The cyclical responses to histamine, 2-methylhistamine, or dimaprit were unaltered by the H1 blocker pyrilamine, were reduced by the H2 antagonist cimetidine, and were abolished by the neuronal blocker tetrodotoxin. Blockade of prostaglandin formation with piroxicam did not prevent the recurrent cycles. The recurrent cycles were inhibited by the chloride transport blocker bumetanide and by removal of chloride ions. Our results demonstrate that histamine mediates prolonged cyclical chloride secretion in the guinea pig distal colon by activating H2 receptors on submucosal neurons involved in regulation of epithelial chloride transport.

Substance P-evoked Cl−secretion in guinea pig distal colonic epithelia: interaction with PGE2

2002 ◽  
Vol 283 (2) ◽  
pp. G347-G356 ◽  
Author(s):  
Yutaka Hosoda ◽  
Shin-Ichiro Karaki ◽  
Yukiko Shimoda ◽  
Atsukazu Kuwahara
Keyword(s):  
Substance P ◽  
Guinea Pig ◽  
Imaging System ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Distal Colon ◽  
Intracellular Camp ◽  
Bathing Solution ◽  
Flux Chamber ◽  
Intracellular Ca

Interaction between substance P (SP) and PGE2on Cl−secretion in the guinea pig distal colonic epithelia was investigated. A short-circuit current ( Isc) was measured as an index of ion transport. Mucosa preparations deprived of muscle and submucosa of distal colon were mounted in the Ussing flux chamber and treated with TTX and piroxicam to remove the influences of neuronal activity and endogenous PG synthesis, respectively. Although SP (10−7M) itself evoked little increase in Isc, exogenous PGE2concentration dependently enhanced the response of SP. The effect of PGE2on the SP-evoked response was mimicked by forskolin and 8-bromoadenosine cAMP. Depletion of Ca2+from the bathing solution reduced the PGE2-dependent response of SP. Effects of PGE2, SP, and SP in the presence of PGE2on intracellular Ca2+concentration ([Ca2+]i) in isolated crypt cells were measured by the confocal microscope fluorescence imaging system. SP, but not PGE2, temporally evoked an increase in [Ca2+]ibut declined to the baseline within 3 min. A return of the SP-evoked increase in [Ca2+]iwas slower in the presence of PGE2than SP alone. These results suggest that PGE2synergistically enhances SP-evoked Cl−secretion via an interaction between the intracellular cAMP and [Ca2+]iin the epithelial cells. In conclusion, SP and PGE2could cooperatively induce massive Cl−secretion in guinea pig distal colon at epithelial levels.

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