Inward rectification in Limulus ventral photoreceptors

1992 ◽  
Vol 8 (1) ◽  
pp. 19-25 ◽  
Author(s):  
Cynthia L. Phillips ◽  
Juan Bacigalupo ◽  
Peter M. O'Day
Keyword(s):  
Late Phase ◽  
Inward Rectifier ◽  
Inward Rectification ◽  
Voltage Range ◽  
Inward Current ◽  
Intracellular Ca ◽  
Dynamic Voltage ◽  
Inward Currents ◽  
Two Phases ◽  
Inwardly Rectifying

AbstractWe examined inward rectification in Limulus ventral photoreceptors using the two-microelectrode voltage clamp. Hyperpolarization in the dark induced an inward current whose magnitude was distinctly dependent on extracellular K+ concentration, [K+0]. The [K+0] dependence resembled the characteristic [K+0] dependence of other inward rectifiers. The inward current was not dependent on extracellular Ca2+ or Na+, and it was unaffected by intracellular injection of Cl−. The hyperpolarization induced currents had two phases, an early nearly instantaneous phase and a slowly developing late phase. The currents were sensitive to extracellular barium and cesium. In voltage-pulse experiments, the magnitudes of the inwardly rectifying currents were variable from cell to cell, with some cells exhibiting negligible inward currents. Large hyperpolarizations (to membrane potentials more negative than about – 140 mV) caused unstable inward current recordings, irreversible desensitization, and irreversible elevation of intracellular Ca2+ concentration. The inward rectifier provides negative feedback by tending to depolarize the cell (with inward current) in response to hyperpolarization. We suggest that the inward rectifier reduces the amount of hyperpolarization that would otherwise be generated by electrogenic processes. This feature would restrict the dynamic voltage range of the photoreceptors at very hyperpolarized potentials.

Inward rectifier K+ currents in smooth muscle cells from rat coronary arteries: block by Mg2+, Ca2+, and Ba2+

1996 ◽  
Vol 271 (2) ◽  
pp. H696-H705 ◽  
Author(s):  
B. E. Robertson ◽  
A. D. Bonev ◽  
M. T. Nelson
Keyword(s):  
Smooth Muscle ◽  
Membrane Potential ◽  
Coronary Artery ◽  
Smooth Muscle Cells ◽  
Coronary Arteries ◽  
Inward Rectifier ◽  
Inward Rectification ◽  
K Channels ◽  
Inward Currents ◽  
K Currents

Inward rectifier K+ channels have been implicated in the control of membrane potential and external K(+)-induced dilations of small coronary arteries. To identify and characterize inward rectifier K+ currents in coronary artery smooth muscle, whole cell K+ currents in smooth muscle cells enzymatically isolated from rat coronary (septal) arteries (diameters, 100-150 microns) were measured in the conventional and perforated configurations of the patch-clamp technique. Ba(2+)-sensitive, whole cell K+ current-voltage relationships exhibited inward rectification. Blockers of Ca(2+)-activated K+ channels (1 mM tetraethylammonium ion), ATP-sensitive K+ channels (10 microM glibenclamide), and voltage-dependent K+ channels (1 mM 4-aminopyridine) in smooth muscle did not affect inward rectifier K+ currents. The nonselective K+ channel inhibitor phencyclidine (100 microM) reduced inward rectifier K+ currents by approximately 50%. External Ba2+ reduced inward currents, with membrane potential hyperpolarization increasing inhibition. The half-inhibition constant for Ba2+ was 2.1 microM at -60 mV, decreasing e-fold for a 25-mV hyperpolarization. External Cs+ also blocked inward rectifier K+ currents, with the half-inhibition constant for Cs+ of 2.9 mM at -60 mV. External Ca2+ and Mg2+ reduced inward rectifier K+ currents. At -60 mV, Ca2+ and Mg2+ (1 mM) reduced inward currents by 33 and 21%, respectively. Inward rectification was not affected by dialysis of the cell's interior with a nominally Ca(2+)- and Mg(2+)-free solution. These findings indicate that inward rectifier K+ channels exist in coronary artery smooth muscle and that Ba2+ may be a useful probe for the functional role of inward rectifier K+ channels in coronary arteries.

Potassium channels regulate tone in rat pulmonary veins

2001 ◽  
Vol 280 (6) ◽  
pp. L1138-L1147 ◽  
Author(s):  
Evangelos D. Michelakis ◽  
E. Kenneth Weir ◽  
Xichen Wu ◽  
Ali Nsair ◽  
Ross Waite ◽  
...  
Keyword(s):  
Low Dose ◽  
Venous Return ◽  
Pulmonary Veins ◽  
Outward Current ◽  
Inward Rectifier ◽  
Inward Current ◽  
Voltage Gated ◽  
Disease States ◽  
Inward Currents ◽  
Vascular Beds

Intrapulmonary veins (PVs) contribute to pulmonary vascular resistance, but the mechanisms controlling PV tone are poorly understood. Although smooth muscle cell (SMC) K+ channels regulate tone in most vascular beds, their role in PV tone is unknown. We show that voltage-gated (KV) and inward rectifier (Kir) K+ channels control resting PV tone in the rat. PVs have a coaxial structure, with layers of cardiomyocytes (CMs) arrayed externally around a subendothelial layer of typical SMCs, thus forming spinchterlike structures. PVCMs have both an inward current, inhibited by low-dose Ba2+, and an outward current, inhibited by 4-aminopyridine. In contrast, PVSMCs lack inward currents, and their outward current is inhibited by tetraethylammonium (5 mM) and 4-aminopyridine. Several KV, Kir, and large-conductance Ca2+-sensitive K+channels are present in PVs. Immunohistochemistry showed that Kir channels are present in PVCMs and PV endothelial cells but not in PVSMCs. We conclude that K+ channels are present and functionally important in rat PVs. PVCMs form sphincters rich in Kir channels, which may modulate venous return both physiologically and in disease states including pulmonary edema.

Acetylcholine reduces inward rectification on thymus-derived macrophage cells in culture

1987 ◽  
Vol 65 (3) ◽  
pp. 348-351 ◽  
Author(s):  
F. Moody-Corbett ◽  
P. Brehm
Keyword(s):  
Outward Current ◽  
Inward Rectifier ◽  
Cholinergic Innervation ◽  
Equilibrium Potential ◽  
Inward Rectification ◽  
Inward Current ◽  
Carbon Particles ◽  
Macrophage Cells ◽  
Anomalous Rectifier ◽  
Cell Current

Cultures prepared from dissociated rat thymus were examined 1–2 weeks after plating. Macrophage cells were identified by their adherence, morphological appearance, and ability to phagocytize carbon particles or heat-inactivated Staphylococcus aureus. Whole cell current recordings from macrophage cells revealed an inward current at potentials more negative than the equilibrium potential for potassium and an outward current at potentials more positive than −40 mV in normal recording solution. Acetylcholine or muscarine caused a reduction in inward current but did not alter the outward current. The inward current and acetylcholine effect were seen at less negative potentials by decreasing the potassium equilibrium potential and both were blocked by the addition of cesium to the external recording solution. These results indicated that the inward current was mediated by potassium through the inward or anomalous rectifier. Physiologically, the action of acetylcholine on the inward rectifier of these macrophage cells may be mediated by cholinergic innervation of the thymus.

scholarly journals Potentiated L-type Ca2+ Channels Rectify

2003 ◽  
Vol 121 (6) ◽  
pp. 541-550 ◽  
Author(s):  
Valérie Leuranguer ◽  
Robert T. Dirksen ◽  
Kurt G. Beam
Keyword(s):  
Outward Current ◽  
Complete Removal ◽  
Inward Rectification ◽  
Wild Type ◽  
Strong Suppression ◽  
Inward Current ◽  
Outward Currents ◽  
Inward Currents ◽  
Bayk 8644 ◽  
Ca2 Channels

Strong depolarization and dihydropyridine agonists potentiate inward currents through native L-type Ca2+ channels, but the effect on outward currents is less clear due to the small size of these currents. Here, we examined potentiation of wild-type α1C and two constructs bearing mutations in conserved glutamates in the pore regions of repeats II and IV (E2A/E4A-α1C) or repeat III (E3K-α1C). With 10 mM Ca2+ in the bath and 110 mM Cs+ in the pipette, these mutated channels, expressed in dysgenic myotubes, produced both inward and outward currents of substantial amplitude. For both the wild-type and mutated channels, we observed strong inward rectification of potentiation: strong depolarization had little effect on outward tail currents but caused the inward tail currents to be larger and to decay more slowly. Similarly, exposure to DHP agonist increased the amplitude of inward currents and decreased the amplitude of outward currents through both E2A/E4A-α1C and E3K-α1C. As in the absence of drug, strong depolarization in the presence of dihydropyridine agonist had little effect on outward tail currents but increased the amplitude and slowed the decay of inward tail currents. We tested whether cytoplasmic Mg2+ functions as the blocking particle responsible for the rectification of potentiated L-type Ca2+ channels. However, even after complete removal of cytoplasmic Mg2+, (−)BayK 8644 still potentiated inward current and partially blocked outward current via E2A/E4A-α1C. Although zero Mg2+ did not reveal potentiation of outward current by DHP agonist, it did have two striking effects, (a) a strong suppression of decay of both inward and outward currents via E2A/E4A-α1C and (b) a nearly complete elimination of depolarization-induced potentiation of inward tail currents. These results can be explained by postulating that potentiation exposes a binding site in the pore to which an intracellular blocking particle can bind and produce inward rectification of the potentiated channels.

Role of arachidonic acid in depolarization-induced modulation of ion currents in Aplysia giant neurons

1990 ◽  
Vol 64 (2) ◽  
pp. 341-350 ◽  
Author(s):  
R. O. Carlson ◽  
I. B. Levitan
Keyword(s):  
Arachidonic Acid ◽  
Calcium Influx ◽  
Reversal Potential ◽  
Inward Current ◽  
Inward Currents ◽  
Pharmacologic Agents ◽  
Inwardly Rectifying ◽  
Aplysia Neuron ◽  
External Calcium

1. The effects of membrane depolarization on inward currents subsequently elicited by hyperpolarization were studied with the use of two-electrode, voltage-clamp techniques in the giant neurons LP1 and R2 of Aplysia. 2. Several successive sets of brief depolarizing pulses, or bursts, were used to depolarize the giant neurons. Two distinct inward currents elicited by hyperpolarization were found to be altered after these sets of depolarizing pulses. These currents were distinguished by their voltage dependence, reversal potential, and sensitivity to 1 mM BaCl2. One of the inward currents was increased after depolarization. It was outwardly rectifying, reversed at -50 mV, and not blocked by Ba2+, suggesting it was a chloride current (ICl). The other inward current, which was decreased after depolarization, was inwardly rectifying, reversed at -70 mV, and completely inhibited by Ba2+. These are characteristics of the inwardly rectifying potassium current (IR), a current previously described to be inhibited after depolarization in Aplysia neuron R 15. Depolarization typically increased the putative ICl and decreased IR for minutes, with the decrease in IR consistently outlasting the increase in an initial brief net increase in inward current followed by a long-lasting decrease. 3. Several criteria suggest arachidonic acid (AA) may mediate depolarization-induced modulation of IR. Previously, free AA has been shown to constitutively inhibit IR in the resting state. Also, depolarization has been reported to stimulate liberation of AA from storage in Aplysia ganglia. Consistent with previous results in neuron R 15, depolarization-induced modulation of IR in giant neurons was dependent on external calcium. Indomethacin and 4-bromophenacylbromide (BPB), pharmacologic agents that activate IR through inhibition of AA turnover, altered the effect of depolarization on IR. In contrast serotonin (5HT), which activates IR through adenosine 3',5'-cyclic monophosphate (cAMP), did not alter the effect of depolarization. Also, extended perfusion with bovine serum albumin (BSA), which strips AA from lipid storage in neurons, decreased the depolarization-induced modulation of IR. We conclude that the calcium influx accompanying depolarization activates the phospholipase responsible for liberation of AA from phospholipid, and the liberated AA then acts to inhibit IR. The molecular mechanism of this AA-mediated inhibition remains to be determined. 4. Depolarization-induced modulation of ICl was also dependent on external calcium but was not affected by BPB and only slightly decreased with indomethacin. This suggested AA was probably not involved in this modulation. However, 5HT opposed the modulation of IC1 induced by previous depolarization, suggesting cAMP may be involved in this effect of depolarization.

scholarly journals Electrostatics in the Cytoplasmic Pore Produce Intrinsic Inward Rectification in Kir2.1 Channels

2005 ◽  
Vol 126 (6) ◽  
pp. 551-562 ◽  
Author(s):  
Shih-Hao Yeh ◽  
Hsueh-Kai Chang ◽  
Ru-Chi Shieh
Keyword(s):  
Cell Types ◽  
Inward Rectifier ◽  
Inward Rectification ◽  
Membrane Excitability ◽  
Channel Inhibition ◽  
Charged Residues ◽  
Voltage Dependent ◽  
Inwardly Rectifying ◽  
Kir2.1 Channel ◽  
Positively Charged

Inward rectifier K+ channels are important in regulating membrane excitability in many cell types. The physiological functions of these channels are related to their unique inward rectification, which has been attributed to voltage-dependent block. Here, we show that inward rectification can also be induced by neutral and positively charged residues at site 224 in the internal vestibule of tetrameric Kir2.1 channels. The order of extent of inward rectification is E224K mutant > E224G mutant > wild type in the absence of internal blockers. Mutating the glycines at the equivalent sites to lysines also rendered weak inward rectifier Kir1.1 channels more inwardly rectifying. Also, conjugating positively charged methanethiosulfonate to the cysteines at site 224 induced strong inward rectification, whereas negatively charged methanethiosulfonate alleviated inward rectification in the E224C mutant. These results suggest that charges at site 224 may control inward rectification in the Kir2.1 channel. In a D172N mutant, spermine interacting with E224 and E299 induced channel inhibition during depolarization but did not occlude the pore, further suggesting that a mechanism other than channel block is involved in the inward rectification of the Kir2.1 channel. In this and our previous studies we showed that the M2 bundle crossing and selectivity filter were not involved in the inward rectification induced by spermine interacting with E224 and E299. We propose that neutral and positively charged residues at site 224 increase a local energy barrier, which reduces K+ efflux more than K+ influx, thereby producing inward rectification.

scholarly journals IRK1 Inward Rectifier K+ Channels Exhibit No Intrinsic Rectification

2002 ◽  
Vol 120 (4) ◽  
pp. 539-551 ◽  
Author(s):  
Donglin Guo ◽  
Zhe Lu
Keyword(s):  
Inward Rectifier ◽  
Inward Rectification ◽  
Induced Current ◽  
Experimental Conditions ◽  
Intact Cells ◽  
K Channels ◽  
Inward Current ◽  
High Affinity ◽  
Modest Degree ◽  
Voltage Jump

In intact cells the depolarization-induced outward IRK1 currents undergo profound relaxation so that the steady-state macroscopic I-V curve exhibits strong inward rectification. A modest degree of rectification persists after the membrane patches were perfused with artificial solutions devoid of Mg2+ and polyamines, which has been interpreted as a reflection of intrinsic channel gating and led to the view that inward rectification results from enhancement of the intrinsic gating by intracellular cations rather than simple pore block. Furthermore, IRK1 exhibits significant extracellular K+-sensitive relaxation of its inward current, a feature that has been likened to the C-type inactivation observed in the voltage-activated Shaker K+ channels. We found that both these current relaxations can be accounted for by impurities in some common constituents of recording solutions, such as residual hydroxyethylpiperazine in HEPES and ethylenediamine in EDTA. Therefore, inherently, IRK1 channels are essentially ohmic at the macroscopic level, and the voltage jump–induced current relaxations do not reflect IRK1 gating but the unusually high affinity of its pore for cations. Furthermore, our study helps define the optimal experimental conditions for studying IRK1.

Inwardly rectifying K+ channels in esophageal smooth muscle

2000 ◽  
Vol 279 (5) ◽  
pp. G951-G960 ◽  
Author(s):  
Junzhi Ji ◽  
Anne Marie F. Salapatek ◽  
Nicholas E. Diamant
Keyword(s):  
Membrane Potential ◽  
Longitudinal Muscle ◽  
Muscle Cells ◽  
Equilibrium Potential ◽  
Inward Rectification ◽  
Resting Membrane Potential ◽  
Patch Clamp Technique ◽  
Membrane Hyperpolarization ◽  
Inward Current ◽  
Inwardly Rectifying

The whole cell patch-clamp technique was used to investigate whether there were inwardly rectifying K+(Kir) channels in the longitudinal muscle of cat esophagus. Inward currents were observable on membrane hyperpolarization negative to the K+ equilibrium potential ( E k) in freshly isolated esophageal longitudinal muscle cells. The current-voltage relationship exhibited strong inward rectification with a reversal potential ( E rev) of −76.5 mV. Elevation of external K+ increased the inward current amplitude and positively shifted its E rev after the E k, suggesting that potassium ions carry this current. External Ba2+ and Cs+ inhibited this inward current, with hyperpolarization remarkably increasing the inhibition. The IC50 for Ba2+ and Cs+ at −60 mV was 2.9 and 1.6 mM, respectively. Furthermore, external Ba2+ of 10 μM moderately depolarized the resting membrane potential of the longitudinal muscle cells by 6.3 mV while inhibiting the inward rectification. We conclude that Kir channels are present in the longitudinal muscle of cat esophagus, where they contribute to its resting membrane potential.

P2X2 receptors are essential for [Ca2+]i increases in response to ATP in cultured rat myenteric neurons

2005 ◽  
Vol 289 (5) ◽  
pp. G935-G948 ◽  
Author(s):  
Toshio Ohta ◽  
Akane Kubota ◽  
Matsuka Murakami ◽  
Ken-ichi Otsuguro ◽  
Shigeo Ito
Keyword(s):  
Rank Order ◽  
Reversal Potential ◽  
Specific Protein ◽  
Inward Rectification ◽  
Channel Blockers ◽  
Patch Clamp Technique ◽  
Myenteric Neurons ◽  
Whole Cell Patch Clamp ◽  
Intracellular Ca ◽  
Inward Currents

We characterized ATP-induced changes in intracellular Ca2+ concentration ([Ca2+]i) and membrane current in cultured rat myenteric neurons using ratiometric Ca2+ imaging with fura-2 and the whole cell patch-clamp technique, respectively. Neuronal cells were functionally identified by [Ca2+]i responses to high K+ and nicotine, which occurred only in cells positive for neuron-specific protein gene product 9.5 immunoreactivity. ATP evoked a dose-dependent increase of [Ca2+]i that was greatly decreased by the removal of extracellular Ca2+ concentration ([Ca2+]o). The amplitude of the [Ca2+]i response to ATP was reduced by half in the presence of voltage-dependent Ca2+ channel blockers. In [Ca2+]o-free solution, ATP produced a small transient rise in [Ca2+]i similar to that induced by P2Y agonists. At −60 mV, ATP evoked a slowly inactivating inward current that was suppressed by the removal of extracellular Na+ concentration. The current-voltage relation for ATP showed an inward rectification with the reversal potential of about 0 mV. The apparent rank order of potency for the purinoceptor agonist-induced increases of [Ca2+]i was ATP ≥ adenosine 5′- O-3-triphosphate ≥ CTP ≥ 2-methylthio-ATP > benzoylbenzoyl-ATP. A similar potency order was obtained with current responses to these agonists. P2 antagonists inhibited inward currents induced by ATP. Ca2+ and Mg2+ suppressed the ATP-induced current, and Zn2+, Cu2+, and protons potentiated it. RT-PCR and immunocytochemical studies showed the expression of P2X2 receptors in cultured rat myenteric neurons. These results suggest that ATP mainly activates ionotropic P2X2 receptors, resulting in a [Ca2+]i increase dependent on [Ca2+]o in rat myenteric neurons. A small part of the ATP-induced [Ca2+]i increase may be also mediated via a P2Y receptor-related mechanism.

scholarly journals Locale and chemistry of spermine binding in the archetypal inward rectifier Kir2.1

2010 ◽  
Vol 135 (5) ◽  
pp. 495-508 ◽  
Author(s):  
Harley T. Kurata ◽  
Emily A. Zhu ◽  
Colin G. Nichols
Keyword(s):  
Binding Site ◽  
Voltage Dependence ◽  
Inward Rectifier ◽  
Selectivity Filter ◽  
Inward Rectification ◽  
Before And After ◽  
Voltage Dependent ◽  
Inwardly Rectifying ◽  
Kinetics Of

Polyamine block of inwardly rectifying potassium (Kir) channels underlies their steep voltage dependence observed in vivo. We have examined the potency, voltage dependence, and kinetics of spermine block in dimeric Kir2.1 constructs containing one nonreactive subunit and one cysteine-substituted subunit before and after modification by methanethiosulfonate (MTS) reagents. At position 169C (between the D172 “rectification controller” and the selectivity filter), modification by either 2-aminoethyl MTS (MTSEA) or 2-(trimethylammonium)ethyl MTS (MTSET) reduced the potency and voltage dependence of spermine block, consistent with this position overlapping the spermine binding site. At position 176C (between D172 and the M2 helix bundle crossing), modification by MTSEA also weakened spermine block. In contrast, MTSET modification of 176C dramatically slowed the kinetics of spermine unblock, with almost no effect on potency or voltage dependence. The data are consistent with MTSET modification of 176C introducing a localized barrier in the inner cavity, resulting in slower spermine entry into and exit from a “deep” binding site (likely between the D172 rectification controller and the selectivity filter), but leaving the spermine binding site mostly unaffected. These findings constrain the location of deep spermine binding that underlies steeply voltage-dependent block, and further suggest important chemical details of high affinity binding of spermine in Kir2.1 channels—the archetypal model of strong inward rectification.

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