scholarly journals Modulation of electroneutral Na transport in sheep rumen epithelium by luminal ammonia

2005 ◽  
Vol 289 (3) ◽  
pp. G508-G520 ◽  
Author(s):  
Khalid Abdoun ◽  
Friederike Stumpff ◽  
Katarina Wolf ◽  
Holger Martens
Keyword(s):  
Patch Clamp ◽  
Ussing Chamber ◽  
Proton Exchange ◽  
Na Transport ◽  
Patch Clamp Technique ◽  
Cytosolic Ph ◽  
Rumen Epithelium ◽  
Joint Application ◽  
Sodium Proton Exchange ◽  
Sheep Rumen

Ammonia is an abundant fermentation product in the forestomachs of ruminants and the intestine of other species. Uptake as NH3 or NH4+ should modulate cytosolic pH and sodium-proton exchange via Na+/H+ exchanger (NHE). Transport rates of Na+, NH4+, and NH3 across the isolated rumen epithelium were studied at various luminal ammonia concentrations and pH values using the Ussing chamber method. The patch-clamp technique was used to identify an uptake route for NH4+. The data show that luminal ammonia inhibits electroneutral Na transport at pH 7.4 and abolishes it at 30 mM ( P < 0.05). In contrast, at pH 6.4, ammonia stimulates Na transport ( P < 0.05). Flux data reveal that at pH 6.4, ∼70% of ammonia is absorbed in the form of NH4+, whereas at pH 7.4, uptake of NH3 exceeds that of NH4+ by a factor of approximately four. The patch-clamp data show a quinidine-sensitive permeability for NH4+ and K+ but not Na+. Conductance was 135 ± 12 pS in symmetrical NH4Cl solution (130 mM). Permeability was modulated by the concentration of permeant ions, with PK > PNH4 at high and PNH4 > PK at lower external concentrations. Joint application of both ions led to anomalous mole fraction effects. In conclusion, the luminal pH determines the predominant form of ammonia absorption from the rumen and the effect of ammonia on electroneutral Na transport. Protons that enter the cytosol through potassium channels in the form of NH4+ stimulate and nonionic diffusion of NH3 blocks NHE, thus contributing to sodium transport and regulation of pH.

scholarly journals Evidence for NHE3-mediated Na transport in sheep and bovine forestomach

2011 ◽  
Vol 301 (2) ◽  
pp. R313-R319 ◽  
Author(s):  
Imtiaz Rabbani ◽  
Christiane Siegling-Vlitakis ◽  
Bardhyl Noci ◽  
Holger Martens
Keyword(s):  
Ussing Chamber ◽  
Specific Inhibitor ◽  
Transepithelial Transport ◽  
Rt Pcr ◽  
Na Transport ◽  
Rumen Epithelium ◽  
Transepithelial Na Transport ◽  
High Concentrations ◽  
Chamber Studies ◽  
Nhe3 Inhibitor

Na absorption across the cornified, multilayered, and squamous rumen epithelium is mediated by electrogenic amiloride-insensitive transport and by electroneutral Na transport. High concentrations of amiloride (>100 μM) inhibit Na transport, indicating Na+/H+ exchange (NHE) activity. The underlying NHE isoform for transepithelial Na absorption was characterized by mucosal application of the specific inhibitor HOE642 for NHE1 and S3226 for NHE3 in Ussing chamber studies with isolated epithelia from bovine and sheep forestomach. S3226 (1 μM; NHE3 inhibitor) abolished electroneutral Na transport under control conditions and also the short-chain fatty acid-induced increase of Na transport via NHE. However, HOE642 (30 μM; NHE1 inhibitor) did not change Na transport rates. NHE3 was immunohistochemically localized in membranes of the upper layers toward the lumen. Expression of NHE1 and NHE3 has been previously demonstrated by RT-PCR, and earlier experiments with isolated rumen epithelial cells have shown the activity of both NHE1 and NHE3. Obviously, both isoforms are involved in the regulation of intracellular pH, pHi. However, transepithelial Na transport is only mediated by apical uptake via NHE3 in connection with extrusion of Na by the basolaterally located Na-K-ATPase. The missing involvement of NHE1 in transepithelial Na transport suggests that the proposed “job sharing” in epithelia between these two isoforms probably also applies to forestomach epithelia: NHE3 for transepithelial transport and NHE1 for, among others, pHi and volume regulation.

Effect of ammonia on Na+ transport across isolated rumen epithelium of sheep is diet dependent

2003 ◽  
Vol 90 (4) ◽  
pp. 751-758 ◽  
Author(s):  
Khalid Abdoun ◽  
Katarina Wolf ◽  
Gisela Arndt ◽  
Holger Martens
Keyword(s):  
Cellular Uptake ◽  
Ussing Chamber ◽  
Ammonium Ion ◽  
Na Transport ◽  
Adaptation Mechanism ◽  
Rumen Epithelium ◽  
Ruminal Epithelium ◽  
Positive Effect ◽  
Chamber Technique

The cellular uptake of ammonia affects the intracellular pH (pHi) of polar and non-polar cells. A predominant uptake of NH3 and its intra-cellular protonation tend to alkalinise the cytoplasm, whereas a predominant uptake of NH4+ acidifies the cytoplasm by reversing this reaction. Hence, the well-known absorption of ammonia across the rumen epithelium probably causes a change in the pHi. The magnitude and direction of this change in pHi (acid or alkaline) depends on the relative transport rates of NH3 and NH4+. Consequently, the intra-cellular availability of protons will influence the activity of the Na+-H+ exchanger, which could affect transepithelial Na+ transport. The aim of the present study has been to test this possible interaction between ruminal ammonia concentrations and Na+ transport. The term ammonia is used to designate the sum of the protonated (NH4+) and unprotonated (NH3) forms. Isolated ruminal epithelium of sheep was investigated by using the Ussing-chamber technique in vitro. The present results indicate that ammonia inhibits Na+ transport across the rumen epithelium of hay-fed sheep, probably by binding intracellular protons and thus inhibiting Na+-H+ exchange. By contrast, ammonia stimulates Na+ transport in concentrate-fed and urea-fed sheep, which develop an adaptation mechanism in the form of an increased metabolism of ammonia in the rumen mucosa and/or an increased permeability of rumen epithelium to the charged ammonium ion (NH4+). Intracellular dissociation of NH4+ increases the availability of protons, which stimulate Na+ –H+ exchange. This positive effect of ruminal ammonia on Na+ absorption may significantly contribute to the regulation of osmotic pressure of the ruminal fluid, because intraruminal ammonia concentrations up to 40 mmol/l have been reported.

Luminal hyperosmolarity decreases Na transport and impairs barrier function of sheep rumen epithelium

2005 ◽  
Vol 175 (8) ◽  
pp. 575-591 ◽  
Author(s):  
Monika Schweigel ◽  
Markus Freyer ◽  
Sabine Leclercq ◽  
Benjamin Etschmann ◽  
Ulrike Lodemann ◽  
...  
Keyword(s):  
Barrier Function ◽  
Na Transport ◽  
Rumen Epithelium ◽  
Sheep Rumen

Vasorelaxing Activity of R-(−)-3′-Hydroxy-2,4,5-trimethoxydalbergiquinol from Dalbergia tonkinensis: Involvement of Smooth Muscle CaV1.2 Channels

Planta Medica ◽  
2020 ◽  
Vol 86 (04) ◽  
pp. 284-293 ◽  
Author(s):  
Nguyen Manh Cuong ◽  
Ninh The Son ◽  
Ngu Truong Nhan ◽  
Pham Ngoc Khanh ◽  
Tran Thu Huong ◽  
...  
Keyword(s):  
Patch Clamp ◽  
Antihypertensive Drugs ◽  
Docking Simulation ◽  
Mechanical Activity ◽  
Dependent Manner ◽  
Vascular Effects ◽  
Patch Clamp Technique ◽  
Spasmolytic Activity ◽  
Cav1.2 Channels ◽  
Dalbergia Tonkinensis

Abstract Dalbergia species heartwood, widely used in traditional medicine to treat various cardiovascular diseases, might represent a rich source of vasoactive agents. In Vietnam, Dalbergia tonkinensis is an endemic tree. Therefore, the aim of the present work was to investigate the vascular activity of R-(−)-3′-hydroxy-2,4,5-trimethoxydalbergiquinol isolated from the heartwood of D. tonkinensis and to provide circular dichroism features of its R absolute configuration. The vascular effects of R-(−)-3′-hydroxy-2,4,5-trimethoxydalbergiquinol were assessed on the in vitro mechanical activity of rat aorta rings, under isometric conditions, and on whole-cell Ba2+ currents through CaV1.2 channels (IBa1.2) recorded in single, rat tail main artery myocytes by means of the patch-clamp technique. R-(−)-3′-Hydroxy-2,4,5-trimethoxydalbergiquinol showed concentration-dependent, vasorelaxant activity on both endothelium-deprived and endothelium intact rings precontracted with the α 1 receptor agonist phenylephrine. Neither the NO (nitric oxide) synthase inhibitor Nω-nitro-L-arginine methyl ester nor the cyclooxygenase inhibitor indomethacin affected its spasmolytic activity. R-(−)-3′-Hydroxy-2,4,5-trimethoxydalbergiquinol-induced vasorelaxation was antagonized by (S)-(−)-Bay K 8644 and unaffected by tetraethylammonium plus glibenclamide. In patch-clamp experiments, R-(−)-3′-hydroxy-2,4,5-trimethoxydalbergiquinol inhibited IBa1.2 in a concentration-dependent manner and significantly decreased the time constant of current inactivation. R-(−)-3′-Hydroxy-2,4,5-trimethoxydalbergiquinol likely stabilized the channel in its closed state, as suggested by molecular modelling and docking simulation to the CaV1.2 channel α 1c subunit. In conclusion, D. tonkinensis species may represent a source of agents potentially useful for the development of novel antihypertensive drugs.

Sodium/Proton Exchange in the Maintenance of Intracellular pH in FRTL-5 Thyroid Cells*

Endocrinology ◽  
1988 ◽  
Vol 123 (4) ◽  
pp. 1705-1711 ◽  
Author(s):  
CLAUDIO MARCOCCI ◽  
EVELYN F. GROLLMAN
Keyword(s):  
Intracellular Ph ◽  
Proton Exchange ◽  
Thyroid Cells ◽  
Sodium Proton Exchange

scholarly journals Ca2+-activated Cl− current in sheep lymphatic smooth muscle

2000 ◽  
Vol 279 (5) ◽  
pp. C1327-C1335 ◽  
Author(s):  
H. M. Toland ◽  
K. D. McCloskey ◽  
K. D. Thornbury ◽  
N. G. McHale ◽  
M. A. Hollywood
Keyword(s):  
Smooth Muscle ◽  
Carboxylic Acid ◽  
Patch Clamp ◽  
Action Potentials ◽  
Channel Blocker ◽  
Equilibrium Potential ◽  
Plateau Phase ◽  
Current Clamp ◽  
Patch Clamp Technique ◽  
Perforated Patch Clamp

Freshly dispersed sheep mesenteric lymphatic smooth muscle cells were studied at 37°C using the perforated patch-clamp technique with Cs+- and K+-filled pipettes. Depolarizing steps evoked currents that consisted ofl-type Ca2+ [ I Ca(L)] current and a slowly developing current. The slow current reversed at 1 ± 1.5 mV with symmetrical Cl− concentrations compared with 23.2 ± 1.2 mV ( n = 5) and −34.3 ± 3.5 mV ( n = 4) when external Cl− was substituted with either glutamate (86 mM) or I− (125 mM). Nifedipine (1 μM) blocked and BAY K 8644 enhanced I Ca(L), the slow-developing sustained current, and the tail current. The Cl− channel blocker anthracene-9-carboxylic acid (9-AC) reduced only the slowly developing inward and tail currents. Application of caffeine (10 mM) to voltage-clamped cells evoked currents that reversed close to the Cl− equilibrium potential and were sensitive to 9-AC. Small spontaneous transient depolarizations and larger action potentials were observed in current clamp, and these were blocked by 9-AC. Evoked action potentials were triphasic and had a prominent plateau phase that was selectively blocked by 9-AC. Similarly, fluid output was reduced by 9-AC in doubly cannulated segments of spontaneously pumping sheep lymphatics, suggesting that the Ca2+-activated Cl− current plays an important role in the electrical activity underlying spontaneous activity in this tissue.

Pancreatic beta-cell electrical activity: the role of anions and the control of pH

1983 ◽  
Vol 244 (3) ◽  
pp. C188-C197 ◽  
Author(s):  
G. T. Eddlestone ◽  
P. M. Beigelman
Keyword(s):  
Membrane Potential ◽  
Cell Membrane ◽  
Beta Cell ◽  
Pancreatic Beta Cell ◽  
Proton Exchange ◽  
Disulfonic Acid ◽  
Control Mechanisms ◽  
Exchange System ◽  
Sodium Proton Exchange

The influence of chloride on the mouse pancreatic beta-cell membrane potential and the cell membrane mechanisms controlling intracellular pH (pHi) have been investigated using glass microelectrodes to monitor the membrane potential. It has been shown that chloride is distributed passively across the beta-cell membrane such that chloride potential is equal to the membrane potential. Withdrawal of perifusate chloride or bicarbonate and the application of the drugs 4-acetamido-4'-isethiocyanostilbene-2,2'-disulfonic acid (SITS) and probenecid, both blockers of transmembrane anion movement, have been used to establish that a chloride-bicarbonate exchange system is operative in the cell membrane and that it is one of the control mechanisms of pHi. Amiloride, a specific blocker of the transmembrane sodium proton exchange, has been used to demonstrate that this mechanism is also operative in the beta-cell membrane in the control of pHi. The hypothesis that the calcium-activated potassium permeability is proton sensitive at an intracellular site, a fall in pHi causing a fall in permeability and an increase in pHi causing an increase in permeability, has been used to explain many of the effects observed in this study.

Ketamine blocks Ca2+-activated K+ channels in rabbit cerebral arterial smooth muscle cells

2003 ◽  
Vol 285 (3) ◽  
pp. H1347-H1355 ◽  
Author(s):  
Jin Han ◽  
Nari Kim ◽  
Hyun Joo ◽  
Euiyong Kim
Keyword(s):  
Smooth Muscle ◽  
Patch Clamp ◽  
Smooth Muscle Cells ◽  
Cerebral Arteries ◽  
Muscle Cells ◽  
Channel Activity ◽  
Patch Clamp Technique ◽  
Arterial Smooth Muscle ◽  
Clamp Technique ◽  
Arterial Smooth Muscle Cells

Although ketamine and Ca2+-activated K+ (KCa) channels have been implicated in the contractile activity regulation of cerebral arteries, no studies have addressed the specific interactions between ketamine and the KCa channels in cerebral arteries. The purpose of this study was to examine the direct effects of ketamine on KCa channel activities using the patch-clamp technique in single-cell preparations of rabbit middle cerebral arterial smooth muscle. We tested the hypothesis that ketamine modulates the KCa channel activity of the cerebral arterial smooth muscle cells of the rabbit. Vascular myocytes were isolated from rabbit middle cerebral arteries using enzymatic dissociation. Single KCa channel activities of smooth muscle cells from rabbit cerebral arteries were recorded using the patch-clamp technique. In the inside-out patches, ketamine in the micromolar range inhibited channel activity with a half-maximal inhibition of the ketamine conentration value of 83.8 ± 12.9 μM. The Hill coefficient was 1.2 ± 0.3. The slope conductance of the current-voltage relationship was 320.1 ± 2.0 pS between 0 and +60 mV in the presence of ketamine and symmetrical 145 mM K+. Ketamine had little effect on either the voltage-dependency or open- and closed-time histograms of KCa channel. The present study clearly demonstrates that ketamine inhibits KCa channel activities in rabbit middle cerebral arterial smooth muscle cells. This inhibition of KCa channels may represent a mechanism for ketamine-induced cerebral vasoconstriction.

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