共查询到20条相似文献,搜索用时 31 毫秒
1.
G Bkaily M Peyrow T Yamamoto A Sculptoreanu D Jacques N Sperelakis 《Molecular and cellular biochemistry》1988,80(1-2):59-72
The whole-cell voltage clamp technique was used to study the slow inward currents and K+ outward currents in single heart cells of embryonic chick and in rabbit aortic cells. In single heart cells of 3-day-old chick embryo three types of slow inward Na+ currents were found. The kinetics and the pharmacology of the slow INa were different from those of the slow ICa in older embryos. Two types of slow inward currents were found in aortic single cells of rabbit; angiotensin II increased the sustained type and d-cAMP and d-cGMP decreased the slow transient component. Two types of outward K+ currents were found in both aortic and heart cells. Single channel analysis demonstrated the presence of a high single K+ channel conductance in aortic cells. In cardiac and vascular smooth muscles, slow inward currents do share some pharmacological properties, although the regulation of these channels by cyclic nucleotides and several drugs seems to be different. 相似文献
2.
J -F Renaud G. Bkaily M. Benabderrazik D. Jacques N. Sperelakis 《Molecular and cellular biochemistry》1989,80(1-2):73-78
Summary The effects of the Ca2+ agonist Bay K 8644 on outward potassium currents have been studied in single ventricular cells of chick embryo and aortic single cells of rabbit using the whole-cell patch clamp technique. Bay K 8644 was found to increase lK in both heart and aortic single cells. This effect of Bay K 8644 on both muscle was reversed by Mn2+ and blocked by 20 mM TEA. The Bay K 8644 potassium I/V curve of single heart cell had a N shape, which is Ca2+ dependent. These data strongly suggest that Bay K 8644 increases a gK(ca) in both aortic and heart muscle. 相似文献
3.
Tohru Kotera Akira Hashimoto Shunji Ueda Yasunobu Okada 《The Journal of membrane biology》1991,124(1):43-52
Summary Patch-clamp studies of whole-cell ionic currents were carried out in parietal cells obtained by collagenase digestion of the gastric fundus of the guinea pig stomach. Applications of positive command pulses induced outward currents. The conductance became progressively augmented with increasing command voltages, exhibiting an outwardly rectifying current-voltage relation. The current displayed a slow time course for activation. In contrast, inward currents were activated upon hyperpolarizing voltage applications at more negative potentials than the equilibrium potential to K+ (E
K). The inward currents showed time-dependent inactivation and an inwardly rectifying current-voltage relation. Tail currents elicited by voltage steps which had activated either outward or inward currents reversed at nearE
K, indicating that both time-dependent and voltagegated currents were due to K+ conductances. Both outward and inward K+ currents were suppressed by extracellular application of Ba2+, but little affected by quinine. Tetraethylammonium inhibited the outward current without impairing the inward current, whereas Cs+ blocked the inward current but not the outward current. The conductance of inward K+ currents, but not outward K+ currents, became larger with increasing extracellular K+ concentration. A Ca2+-mobilizing acid secretagogue, carbachol, and a Ca2+ ionophore, ionomycin, brought about activation of another type of outward K+ currents and voltage-independent cation currents. Both currents were abolished by cytosolic Ca2+ chelation. Quinine preferentially inhibited this K+ current. It is concluded that resting parietal cells of the guinea pig have two distinct types of voltage-dependent K+ channels, inward rectifier and outward rectifier, and that the cells have Ca2+-activated K+ channels which might be involved in acid secretion under stimulation by Ca2+-mobilizing secretagogues. 相似文献
4.
Ira R. Josephson 《Molecular and cellular biochemistry》1989,80(1-2):21-26
Summary The voltage- and time-dependent properties of whole-cell, multi-channel (outside-out), and single channel inwardly-rectifying K+ currents were studied using adult and neonatal rat, and embryonic chick ventricular myocytes. Inward rectification of the current-voltage relationship was found in the whole-cell and single channel measurements. The steady-state single channel probability of opening decreased with hyperpolarization from EK, as did the mean open time, thereby explaining the time-dependent inactivation of the macroscopic current. Myocytes dialysed with a Mg++-free K+ solution (to remove the property of inward rectification) displayed a quasi-linear current-voltage relationship. The outward K+ currents flowing through the modified inward rectifier channels were able to be blocked by the local anesthetic and anti-arrhythmic agent, lidocaine. 相似文献
5.
Mark Tester 《The Journal of membrane biology》1988,105(1):77-85
Summary The outer membranes of plant cells contain channels which are highly selective for K+. In the giant-celled green algaChara corallina, K+ currents in the plasmalemma were measured during the action potential and when the cell was depolarized to the K+ equilibrium potential in high external K+ concentrations. Currents in both conditions were reduced by externally added tetraethylammonium (TEA+), Ba2+, Na+ and Cs+. In contrast to inhibition by TEA+, the latter three ions inhibited inward K+ current in a voltage-dependent manner, and reduced inward current more than outward. Ba2+ and Na+ also appeared to inhibit outward current in a strongly voltage-dependent manner. The blockade by Cs+ is studied in more detail in the following paper. TEA+ inhibited both inward and outward currents in a largely voltage-independent manner, with an apparentK
D
of about 0.7 to 1.1mm, increasing with increasing external K+. All inhibitors reduced current towards a similar linear leak, suggesting an insensitivity of the background leak inChara to these various K+ channel inhibitors. The selectivity of the channel to various monovalent cations varied depending on the method of measurement, suggesting that ion movement through the K+-selective channel may not be independent. 相似文献
6.
Nicholas Sperelakis Yoshihito Inoue Yusuke Ohya 《Molecular and cellular biochemistry》1992,114(1-2):79-89
Summary Smooth muscle cells normally do not possess fast Na2+ channels, but inward current is carried through two types of Ca2+ channels: slow (L-type) Ca2+ channels and fast (T-type) Ca2+ channels. Using whole-cell voltage clamp of single smooth muscle cells isolated from the longitudinal layer of 18-day pregnant rat uterus, depolarizing pusles, applied from a holding potential of –90 mV, evoked two types of inward current, fast and slow [8]. The fast inward current decayed within 30 ms, depended on [Na]0, and was inhibited by TTX (K0.5 = 27 nM). The slow inward current decayed slowly, was dependent on [Ca]0, and was inhibited by nifedipine. These results suggest that the fast inward current is a fast Na2+ channel current, and that the slow inward current is a Ca2+ channel current was not evident. Thus, the ion channels which generate inward currents in pregnant rat uterine cells are TTX-sensitive fast Na+ channels and dihudropuridine-sensitive slow Ca2+ channels. The number of fast Na+ channels increased during gestation [9]. The averaged current density increased from 0 on day 5, to 0.19 on day 9, to 0.56 on day 14, to 0.90 on day 18, and to 0.86 pA/pF on day 21. This almost linear increase occurs because of an increase in the fraction of cells which possess fast Na2+ channels, and it suggested that the fast Na+ current may be involved in spread of excitation. The Ca2+ channel current density also was higher during the latter half of gestation. These results indicate that the fast Na+ channels and Ca2+ slow channels in myometrium become more numerous as term approaches, and may facilitate parturition. Isoproterenol (beta-agonist) did not affect either ICa(s) or INa(f), whereas Mg2+ (K0.5 of 12 mM) and nifedipine (K0.5 of 3.3 nM) depressed ICa(s). Oxytocin had no effect on INa(f) and actually depressed ICa(s) to a small extect. Therefore, the tocolytic action of beta-agonists cannot be explained by an inhibition of ICa(s), whereas that of Mg2+ can be so explained. The stimulating action of oxytocin on uterine contractions is not due to stimulation of ICa(s). 相似文献
7.
W. Schreibmayer H. A. Tritthart G. Zernig H. M. Piper 《European biophysics journal : EBJ》1985,11(4):259-263
Studies on single K+-channel currents recorded from isolated rat heart muscle cells, in which early repolarization is known to be exceptionally fast, are reported here. A K+-channel which is blocked by TEA (tetraethylammonium) from the inside only has been found.The total open time of the channel, measured in steady-state after activation, indicated outward rectifying properties. The single channel conductance increases with depolarization from 25 pS at-70 mV to 75 pS at+70 mV.Selectivity of the channel has also been measured and it was found that only Rb+ and K+ can permeate the channel, whereas the permeability (P) for Li+, Na+, Cl-, Mg2+, and Ca2+ is less than 0.05 times
.Ba2+ and Cs+ block the channel activity.These results clearly demonstrate the existence of K+-selective outward rectifying conductance pathways in rat ventricular myocytes. 相似文献
8.
Jacques Danielle Bkaily Ghassan Jasmin Gaétan Ménard Daniel Proschek Libuse 《Molecular and cellular biochemistry》1997,176(1-2):249-256
Using the whole-cell voltage-clamp technique, early embryonic tetrodotoxin (TTX) and Mn2+-insensitive slow Na+ current was detected in 10-22 week old fetal human heart cells as well as in 1-day-old and young cardiomyopathic hamster myocytes. This slow Na+ current in both heart cell preparations has the same kinetics and pharmacology. This type of slow Na+ current was absent in heart cells of newborn and young normal hamsters and became less present in myocytes of 19 and 22 week old human heart myocytes. Our results demonstrate that the slow Na+ channel does exist in early fetal human life and this type of channel continues to be functional after birth in myocytes of the hereditary cardiomyopathic hamster. 相似文献
9.
Membrane potential and ionic currents were studied in cultured rabbit retinal pigment epithelial (RPE) cells using whole-cell patch clamp and perforated-patch recording techniques. RPE cells exhibited both outward and inward voltage-dependent currents and had a mean membrane capacitance of 26±12 pF (sd, n=92). The resting membrane potential averaged ?31±15 mV (n=37), but it was as high as ?60 mV in some cells. When K+ was the principal cation in the recording electrode, depolarization-activated outward currents were apparent in 91% of cells studied. Tail current analysis revealed that the outward currents were primarily K+ selective. The most frequently observed outward K+ current was a voltage- and time-dependent outward current (I K) which resembled the delayed rectifier K+ current described in other cells. I K was blocked by tetraethylammonium ions (TEA) and barium (Ba2+) and reduced by 4-aminopyridine (4-AP). In a few cells (3–4%), depolarization to ?50 mV or more negative potentials evoked an outwardly rectifying K+ current (I Kt) which showed more rapid inactivation at depolarized potentials. Inwardly rectifying K+ current (I KI) was also present in 41% of cells. I KI was blocked by extracellular Ba2+ or Cs+ and exhibited time-dependent decay, due to Na+ blockade, at negative potentials. We conclude that cultured rabbit RPE cells exhibit at least three voltage-dependent K+ currents. The K+ conductances reported here may provide conductive pathways important in maintaining ion and fluid homeostasis in the subretinal space. 相似文献
10.
Patch-clamp experiments were performed on satellite glial cells wrapped around sympathetic neurons in the rabbit coeliac ganglion. With the cleaning method used, the glial cells could be kept in place and were directly accessible to the patch-clamp pipettes. Whole-cell recordings showed that glial cells had almost ohmic properties. Their resting potential (–79.1±1.2 mV) was found to be very nearly the same as the K+ reversal potential and 20 mV more negative than that of the neurons they encapsulated. Unitary currents from ionic channels present in the glial membrane were recorded in the cell-attached configuration with pipettes filled with various amounts of K+, Na+ and gluconate. Only K+-selective channels with slight inwardly rectifying properties (in the presence of 150 mM [K+]0) were detected. These channels were active (P
0=0.7–0.8) at the cell resting potential. The channel conductance, but not its opening probability, was dependent on the [K+] in the pipette. Cl–-selective channels (outwardly rectifying and large conductance channels) were detected in excised patches.The properties of the K+ channels (increased inward current with [K+] and detectable outward current at low [K+]) are well suited for siphoning the K+ released by active neurons. 相似文献
11.
UV irradiation has multiple effects on mammalian cells, including modification of ion channel function. The present study
was undertaken to investigate the response of membrane currents in guinea-pig ventricular myocytes to the type A (355, 380
nm) irradiation commonly used in Ca2+ imaging studies. Myocytes configured for whole-cell voltage clamp were generally held at −80 mV, dialyzed with K+-, Na+-free pipette solution, and bathed with K+-free Tyrode’s solution at 22°C. During experiments that lasted for ≈ 35 min, UVA irradiation caused a progressive increase
in slowly-inactivating inward current elicited by 200-ms depolarizations from −80 to −40 mV, but had little effect on background
current or on L-type Ca2+ current. Trials with depolarized holding potential, Ca2+ channel blockers, and tetrodotoxin (TTX) established that the current induced by irradiation was late (slowly-inactivating)
Na+ current (INa). The amplitude of the late inward current sensitive to 100 μM TTX was increased by 3.5-fold after 20–30 min of irradiation. UVA modulation of late INa may (i) interfere with imaging studies, and (ii) provide a paradigm for investigation of intracellular factors likely to
influence slow inactivation of cardiac INa. 相似文献
12.
The Interaction of Na+ and K+ in Voltage-gated Potassium Channels : Evidence for Cation Binding Sites of Different Affinity
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Laszlo Kiss David Immke Joseph LoTurco Stephen J. Korn 《The Journal of general physiology》1998,111(2):195-206
Voltage-gated potassium (K+) channels are multi-ion pores. Recent studies suggest that, similar to calcium channels, competition between ionic species for intrapore binding sites may contribute to ionic selectivity in at least some K+ channels. Molecular studies suggest that a putative constricted region of the pore, which is presumably the site of selectivity, may be as short as one ionic diameter in length. Taken together, these results suggest that selectivity may occur at just a single binding site in the pore. We are studying a chimeric K+ channel that is highly selective for K+ over Na+ in physiological solutions, but conducts Na+ in the absence of K+. Na+ and K+ currents both display slow (C-type) inactivation, but had markedly different inactivation and deactivation kinetics; Na+ currents inactivated more rapidly and deactivated more slowly than K+ currents. Currents carried by 160 mM Na+ were inhibited by external K+ with an apparent IC50 <30 μM. K+ also altered both inactivation and deactivation kinetics of Na+ currents at these low concentrations. In the complementary experiment, currents carried by 3 mM K+ were inhibited by external Na+, with an apparent IC50 of ∼100 mM. In contrast to the effects of low [K+] on Na+ current kinetics, Na+ did not affect K+ current kinetics, even at concentrations that inhibited K+ currents by 40–50%. These data suggest that Na+ block of K+ currents did not involve displacement of K+ from the high affinity site involved in gating kinetics. We present a model that describes the permeation pathway as a single high affinity, cation-selective binding site, flanked by low affinity, nonselective sites. This model quantitatively predicts the anomalous mole fraction behavior observed in two different K+ channels, differential K+ and Na+ conductance, and the concentration dependence of K+ block of Na+ currents and Na+ block of K+ currents. Based on our results, we hypothesize that the permeation pathway contains a single high affinity binding site, where selectivity and ionic modulation of gating occur. 相似文献
13.
Summary Elementary Na+ currents through single cardiac Na+ channels were recorded at –50 mV in cell-attached patches from neonatal rat cardiocytes kept at holding potentials between –100 and –120 mV.Na+ channel activity may occur as burst-like, closely-timed repetitive openings with shut times close to 0.5–0.6 msec, indicating that an individual Na+ channel may reopen several times during step depolarization. A systematic quantiative analysis in 19 cell-attached patches showed that reopening may be quite differently pronounced. The majority, namely 16 patches, contained Na+ channels with a low tendency to reopen. This was evidenced from the average value for the mean number of openings per sequence, 2.5. Strikingly different results were obtained in a second group of three patches. Here, a mean number of openings per sequence of 3.42, 3.72, and 5.68 was found. Ensemble averages from the latter group of patches revealed macroscopic Na+ currents with a biexponential decay phase. Reconstructed Na+ currents from patches with poorly reopening Na+ channels were devoid of a slow decay component. This strongly suggests that reopening may be causally related to slow Na+ inactivation. Poorly pronounced reopening and, consequently, the lack of slow Na+ inactivation could be characteristic features of neonatal cardiac Na+ channels. 相似文献
14.
Summary Human red cells were prepared with various cellular Na+ and K+ concentrations at a constant sum of 156mm. At maximal activation of the K+ conductance,g
K(Ca), the net efflux of K+ was determined as a function of the cellular Na+ and K+ concentrations and the membrane potential,V
m
, at a fixed [K+]ex of 3.5mm.V
m
was only varied from (V
m
–E
K)25 mV and upwards, that is, outside the range of potentials with a steep inward rectifying voltage dependence (Stampe & Vestergaard-Bogind, 1988).g
K(Ca) as a function of cellular Na+ and K+ concentrations atV
m
=–40, 0 and 40 mV indicated a competitive, voltage-dependent block of the outward current conductance by cellular Na+. Since the present Ca2+-activated K+ channels have been shown to be of the multi-ion type, the experimental data from each set of Na+ and K+ concentrations were fitted separately to a Boltzmann-type equation, assuming that the outward current conductance in the absence of cellular Na+ is independent of voltage. The equivalent valence determined in this way was a function of the cellular Na+ concentration increasing from 0.5 to 1.5 as this concentration increased from 11 to 101mm. Data from a previous study of voltage dependence as a function of the degree of Ca2+ activation of the channel could be accounted for in this way as well. It is therefore suggested that the voltage dependence ofg
K(Ca) for outward currents at (V
m
–E
K)>25 25 mV reflects a voltage-dependent Na+ block of the Ca2+-activated K+ channels. 相似文献
15.
The bacterial potassium (K+) channel KcsA provides an attractive model system to study ion permeation behavior in a selective K+-channel. We changed residue at the N-terminal end of the selectivity filter of KcsA (T74V) to its counterpart in inwardly rectifying K+-channels (Kir). The tetramer was found to be stable as unmodified KcsA. Under symmetrical and asymmetrical conditions, Na+ increased the inward current in the virtual absence of K+ however outward currents were nearly abolished which could be recovered upon internal K+ addition. Na+ also drastically increased the channel open time either in the presence or virtual absence of K+. Furthermore, the T74V mutation decreased the internal Ba2+ affinity of the channel possibly by binding to a K+ site in the pore. In additional experiments, another point mutation V76I in T74V mutant was carried out thus the selectivity filter resembled more the selectivity filter of Kir channels. The mutant tetramer was converted into monomers as determined by conventional gel electrophoresis. However, native like gel electrophoresis, Trp fluorescence and acrylamide quenching experiments indicated that this mutant still formed a tetramer and apparently adopted similar folding properties as unmodified KcsA. Single-channel experiments further demonstrated that the channel was selective for K+ over Na+ as Na+ blocked channel currents. These data suggest that single point mutation T74V alters the selectivity filter and allows simultaneous occupancy and conduction of K+ and Na+ probably via ion–ion interaction in the pore. In contrast, both mutations (T74V and V76I) in the same molecule seem to reorganize the pore conformation which controls the overall stability of a selective K+-channel. 相似文献
16.
Palle Christophersen 《The Journal of membrane biology》1991,119(1):75-83
Summary The Ca2+-activated K+ channel of the human red cell membranes was characterized with respect to rectification and selectivity using the patch-clamp technique. In inside-out patches exposed to symmetric solutions of K+, Rb+, and NH
4
+
, respectively, inward rectifyingi-V curves were obtained. The zero current conductances were: K+ (23.5 pS±3.2)>NH
4
+
(14.2 pS±1.2)>Rb+ (11.4 pS±1.8). With low extracellular K+ concentrations (substitution with Na+) the current fluctuations reversed close to the Nernst potential for the K ion and the rectification as well as thei-V slopes decreased. With mixed intracellular solutions of K+ and Na+ enhanced rectification were observed due to a Na+ block of outward currents. From bi-ionic reversal potentials the following permeability sequence (P
K/P
X) was calculated: K+ (1.0)>Rb+ (1.4±0.1)>NH
4
+
(8.5±1.3)>Li+(>50); Na+ (>110); Cs+ (5). Li+, Na+, and Cs+ were not found to carry any current, and only minimum values of the permeability ratios were estimated. Tl+ was permeant, but the permeability and conductance were difficult to quantify, since with this ion the single channel activity was extremely low and the channels seemed to inactivate. The inward rectification in symmetric solutions indicate an asymmetric open channel structure, and the different selectivity sequences based on conductances and permeabilities reflect interionic interactions in the permeation process. 相似文献
17.
Using the two-microelectrode voltage clamp technique in Xenopus laevis oocytes, we estimated Na+-K+-ATPase activity from the dihydroouabain-sensitive current (I
DHO) in the presence of increasing concentrations of tetraethylammonium (TEA+; 0, 5, 10, 20, 40 mm), a well-known blocker of K+ channels. The effects of TEA+ on the total oocyte currents could be separated into two distinct parts: generation of a nonsaturating inward current increasing with negative membrane potentials (V
M) and a saturable inhibitory component affecting an outward current easily detectable at positive V
M. The nonsaturating component appears to be a barium-sensitive electrodiffusion of TEA+ which can be described by the Goldman-Hodgkin-Katz equation, while the saturating component is consistent with the expected blocking effect of TEA+ on K+ channels. Interestingly, this latter component disappears when the Na+-K+-ATPase is inhibited by 10 m DHO. Conversely, TEA+ inhibits a component of I
DHO with a k
d of 25±4 mm at +50 mV. As the TEA+-sensitive current present in I
DHO reversed at –75 mV, we hypothesized that it could come from an inhibition of K+ channels whose activity varies in parallel with the Na+-K+-ATPase activity. Supporting this hypothesis, the inward portion of this TEA+-sensitive current can be completely abolished by the addition of 1 mm Ba2+ to the bath. This study suggests that, in X. laevis oocytes, a close link exists between the Na-K-ATPase activity and TEA+-sensitive K+ currents and indicates that, in the absence of effective K+ channel inhibitors, I
DHO does not exclusively represent the Na+-K+-ATPase-generated current. 相似文献
18.
Extracellular acidification and reduction of extracellular K+ are known to decrease the currents of some voltage-gated potassium channels. Although the macroscopic conductance of WT hKv1.5 channels is not very sensitive to [K+]o at pH 7.4, it is very sensitive to [K+]o at pH 6.4, and in the mutant, H463G, the removal of K+
o virtually eliminates the current at pH 7.4. We investigated the mechanism of current regulation by K+
o in the Kv1.5 H463G mutant channel at pH 7.4 and the wild-type channel at pH 6.4 by taking advantage of Na+ permeation through inactivated channels. Although the H463G currents were abolished in zero [K+]o, robust Na+ tail currents through inactivated channels were observed. The appearnnce of H463G Na+ currents with a slow rising phase on repolarization after a very brief depolarization (2 ms) suggests that channels could
activate directly from closed-inactivated states. In wild-type channels, when intracellular K+ was replaced by NMG+ and the inward Na+ current was recorded, addition of 1 mM K+ prevented inactivation, but changing pH from 7.4 to 6.4 reversed this action. The data support the idea that C-type inactivation
mediated at R487 in Kv1.5 channels is influenced by H463 in the outer pore. We conclude that both acidification and reduction
of [K+]o inhibit Kv1.5 channels through a common mechananism (i.e., by increasing channel inactivation, which occurs in the resting
state or develops very rapidly after activation). 相似文献
19.
Melanie E. M. Kelly S. Jeffrey Dixon Stephen M. Sims 《The Journal of membrane biology》1992,126(2):171-181
Summary Ionic conductances of rabbit osteoclasts were investigated using both whole-cell and cell-attached configurations of the patch-clamp recording technique. The predominant conductance found in these cells was an inwardly rectifying K+ conductance. Whole-cell currents showed an N-shaped current-voltage (I–13;V) relation with inward current activated at potentials negative to EK. When external K+ was varied, I-V curves shifted 53 mV/10-fold change in [K+]out, as predicted for a K+-selective channel. Inward current was blocked by Ba2+ and showed a time-dependent decline at negative potentials, which was reduced in Na+-free external solution. Inward single-channel currents were recorded in the cell-attached configuration. Single-channel currents were identified as inward-rectifier K+ channels based on the following observations: (i) Unitary I-V relations rectified, with only inward current resolved. (ii) Unitary conductance () was 31 pS when recorded in the cell-attached configuration with 140 mm K+ in the pipette and was found to be dependent on [K+]. (iii) Addition of Ba2+ to the pipette solution abolished single-channel events. We conclude that rabbit osteoclasts possess inwardly rectifying K+ channels which give rise to the inward current recorded at negative potentials in the whole-cell configuration. This inwardly rectifying K+ current may be responsible for setting the resting membrane potential and for dissipating electrical potential differences which arise from electrogenic transport of protons across the osteoclast ruffled border.This work was supported by The Arthritis Society and the Medical Research Council of Canada. M.E.M.K. was supported by a fellowship, S.J.D. a development Grant and S.M.S. a scholarship from the Medical Research Council. We thank Dr. Zu Gang Zheng for help with scanning microscopy. 相似文献
20.
The role of the inward K+ rectifier in the repetitive activity at depolarized levels was studied in guinea pig single ventricular myocytes by voltage- and current-clamp methods. In action potentials arrested at the plateau by a depolarizing current, small superimposed hyperpolarizing currents caused much larger voltage displacements than at the resting potential and sometimes induced a regenerative repolarization. Around –20 mV, sub- and suprathreshold repetitive inward currents were found. In the same voltage range, small hyperpolarizing currents reversed their polarity. During depolarizing voltage-clamp ramps, around –20 mV there was a sudden decrease in the outward current (Ins: current underlying the negative slope in the inward K+ rectifier steady state I–V relation). During repolarizing ramps, the reincrease in outward current was smaller and slower. During depolarizing and repolarizing current ramps, sudden voltage displacements showed a similar asymmetry. Repetitive Ins could continue as long as the potential was kept at the level at which they appeared. Depolarizing voltage-clamp steps also caused repetitive Ins and depolarizing current steps induced repetitive slow responses. Cadmium and verapamil reduced Ins amplitude during the depolarizing ramp. BRL 34915 (cromakalim), an opener of the ATP-sensitive K+ channel, eliminated the negative slope and Ins, whereas barium increased Ins frequency (an effect abolished by adding BRL). Depolarization-induced slow responses persisted in an NaCl-Ca-free solution. Thus, the mechanism of repetitive activity at the depolarized level appears to be related to the presence of the negative slope in the inward K+ rectifier I–V relation. 相似文献