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1.
The aim of this study was to measure and compare the profile of rapid delayed rectifier potassium current (IKr) elicited by action potential (AP) waveforms applied to isolated rabbit atrioventricular nodal (AVN) and ventricular myocytes. All measurements were made using whole-cell patch-clamp recordings at 37 degrees C. In AVN myocytes, IKr during voltage steps and slow ramp depolarisations showed "inward rectification" (characteristic for this channel) at positive potentials. The E-4031-sensitive current showed half-maximal activation at -10.8 +/- 0.86 mV, with a slope factor for the activation relation of 6.5 +/- 0.77 mV (n = 7). During AVN APs, IKr rapidly reached a peak after the AP upstroke and remained at similar amplitude until late in AP repolarisation. At the maximum diastolic potential following the AVN AP, a component of IKr remained which decayed during the pacemaker depolarisation, consistent with a role for the current in generating AVN pacemaker activity. In ventricular myocytes IKr was small at the beginning of the AP, and increased slowly during the AP plateau. Measurement of Ba-sensitive-inward rectifier K current (IK1) in ventricular myocytes revealed that IK1 rapidly increased during the final AP repolarisation phase, whilst IKr declined. It is concluded that IKr may participate in both AP repolarisation and the pacemaker depolarisation in AVN cells, whilst in ventricular myocytes, IKr and IK1 participate in controlling early and final AP repolarisation respectively. 相似文献
2.
Muscarine induces two distinct current responses in adrenal chromaffin cells of the guinea-pig 总被引:1,自引:0,他引:1
Muscarine-induced membrane responses were studied in dissociated chromaffin cells of the guinea-pig adrenal medulla, using the whole-cell version of the patch-clamp technique. Bath application of muscarine (1-10 microM) produced two distinct current responses at a holding potential of -40 mV. One is an inward current associated with an increase in current noise. This current response was sustained during stimulation and had a reversal potential of 4.5 +/- 3.4 mV (n = 6) with a negative slope conductance below about -30 mV in 12.5 mM K(+)-containing perfusate. The other is a transient outward current. This was evoked at membrane potentials more positive than -60 mV and completely suppressed by addition of 2 mM TEA to the bath solution, suggesting a possible involvement of the Ca2(+)-dependent K+ channel. Generation of the outward current response was suppressed for at least 60-90 s following 25 s muscarinic stimulation and was facilitated by activation of the nicotinic receptor. The maximum inward current seemed to be produced by 3 microM, whereas the threshold concentration required for generation of the outward current was somewhere between 3 and 10 microM. The outward current was evoked less often in cells treated with 2% collagenase for 1 h than in those treated with 0.2% for 30 min. The results suggest that guinea-pig chromaffin cells have two muscarinic receptors: one is coupled with a cation nonselective channel and the other may be related to a Ca2(+)-dependent K+ channel. 相似文献
3.
Externally applied ATP (100 μM) induced membrane currents in type I and type II vestibular hair cells enzymatically isolated from guinea-pig semicircular canals. In whole-cell voltage-clamp and with 140 mM K+ in the pipette solution, ATP evoked an inwardly directed current in 58% of the cells when held at potentials below −40 mV. In the remainder, external ATP produced an outward current. After block of the K currents, an inward current activated by ATP was revealed at −50 mV. Intracellular Ca2+ levels were monitored using the Ca2+ indicator Fura-2 and were found to rise in both hair cell types in response to ATP. These results strongly suggest that ATP directly controls the entry of Ca2+ into crista hair cells which can then further modulate K+ currents. 相似文献
4.
Ca2+-channel current and its modification by the dihydropyridine agonist BAY k 8644 in isolated smooth muscle cells 总被引:9,自引:0,他引:9
The electrophysiological properties of single smooth muscle cells isolated from the longitudinal layer of the guinea-pig ileum were studied with the whole-cell patch-clamp technique. The finding of resting potentials between -45 and -50 mV and the occurrence of spontaneous electrical activity when K+ was the predominant intracellular cation indicated that the cells were not leaky or hyperpermeable. The existence of an inward Ca2+ current overlapping in time with an outward rectifying K+ current was demonstrated. The latter could be selectively blocked by replacing internal K+ with Cs+ and external Ca2+ with Ba2+. Depolarizations to potentials between -40 and +50 mV evoked time-dependent inward currents, with a maximum peak value between -20 and 0 mV. For depolarizations beyond +50 mV time-dependent outward currents appeared. These currents were inhibited by 0.1 mM CdCl2. The activation of the inward current showed a sigmoidal time course, and the rate of onset of the current increased at more positive potentials. Inactivation could be described by two exponentials. The threshold for activation was about -40 mV, and full activation was reached at 0 mV. Inactivation was complete near 0 mV, whereas the channels were fully available at -80 mV. The fully-activated Ca2+-channel current was strongly voltage dependent. The conductance decreased for potentials close to the reversal potential, and showed rectification for hyperpolarizing potentials. The Ca2+ agonist BAY k 8644 enhanced the Ca2+-channel current without a significant effect on its kinetics. The fully-activated current and the steady-state activation were enhanced in a rather voltage-independent way. 相似文献
5.
6.
A transient outward current (I
to) has been observed in the atrioventricular node (AVN), but its characteristics in Ca-tolerant AVN myocytes have not been
investigated previously. In this study, I
to was measured from Ca-tolerant rabbit AVN myocytes at 37°C, using the whole-cell patch-clamp technique. With interfering currents
inhibited, 500-ms voltage-clamp pulses applied from –80 mV elicited I
to at potentials positive to –30 mV, which increased in magnitude with test potential amplitude. This current was completely
blocked by external application of 5 mM 4-aminopyridine (4-AP). During a command pulse, I
to activated rapidly then inactivated with a bi-exponential time-course. Fast and slow time constants of current inactivation
(τf and τs, respectively) showed voltage dependence. At 0 mV, τf was 14.5±2.7 ms and τs was 112.8±21.2 ms, whilst at +60 mV τf was 6.7±1.1 ms and τs was 63.7±9.2 ms (n=25). The steady-state inactivation relationship showed half-maximal inactivation at –33.8 mV (n=8). Re-activation of I
to after an inactivating pre-pulse showed a bi-exponential time-course of recovery: τ1 was 196±70 ms, and τ2 was 2707±1010 ms (n=6, at –80 mV). Repetitive application of voltage-clamp test pulses showed that I
to inactivation accumulated on repetitive stimulation, but reached a steady state rapidly for a given pulse frequency (0.2–1.0
Hz). AVN I
to was sensitive to the class 1 anti-arrhythmic flecainide (EC50 for peak current of 24 μM), which showed selectivity for the rapidly inactivating current component. Quinidine also inhibited I
to in a dose-dependent fashion, but did not affect the current time-course. Under voltage-clamp conditions, a simulated diastolic
depolarisation from –70 to –45 mV did not significantly reduce I
to amplitude, and under current-clamp conditions 4-AP inhibited spontaneous action potentials. Although this is consistent with
a significant role for I
to in shaping AVN activity, under the conditions of this study 4-AP also partially blocked the ”rapid” delayed rectifier current, I
Kr, and so the effects of 4-AP on action potentials could not be attributed exclusively to its effects on I
to.
Received: 22 October 1998 / Received after revision: 19 January 1999 / Accepted: 20 January 1999 相似文献
7.
Properties of the inwardly rectifying K+ conductance in the toad retinal pigment epithelium. 总被引:1,自引:4,他引:1
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An inwardly rectifying K+ current was analysed in isolated toad retinal pigment epithelial (RPE) cells using the perforated-patch clamp technique. The zero-current potential (Vo) of RPE cells averaged -71 mV when the extracellular K+ concentration ([K+]o) was 2 mM. Increasing [K+]o from 0.5 to 5 mM shifted V0 by +43 mV, indicating a relative K+ conductance (TK) of 0.74. At [K+]o greater than 5 mM, TK decreased to 0.53. Currents were larger in response to hyperpolarizing voltage pulses than depolarizing pulses, indicating an inwardly rectifying conductance. Currents were time independent except in response to voltage pulses to potentials positive to 0 mV, where the outward current decayed with an exponential time course. Both the inwardly rectifying current and the transient outward current were eliminated by the addition of 0.5 mM Ba2+, 5 mM Cs+ or 2 mM Rb+ to the extracellular solution. The current blocked by these ions reversed near the K+ equilibrium potential (EK) over a wide range of [K+]o, indicating a highly selective K+ channel. The current-voltage relationship of the isolated K+ current exhibited mild inward rectification at voltages negative to -20 mV and a negative slope conductance at voltages positive to -20 mV. The Cs(+)- and Ba(2+)-induced blocks of the K+ current were concentration dependent but voltage independent. The apparent dissociation constants were 0.8 mM for Cs+ and 40 microM for Ba2+. The K+ conductance decreased when extracellular Na+ was removed. Increasing [K+]o decreased the K+ chord conductance (gK) at negative membrane potentials. In the physiological voltage range, increasing [K+]o from 2 to 5 mM caused gK to decrease by approximately 25%. We conclude that the inwardly rectifying K+ conductance represents the resting K+ conductance of the toad RPE apical membrane. The unusual properties of this conductance may enhance the ability of the RPE to buffer [K+]o changes that take place in the subretinal space at the transition between dark and light. 相似文献
8.
1. Voltage-dependent currents of untreated (proliferating) and lipopolysaccharide (LPS)-treated rat microglial cells in culture were recorded using the whole-cell patch-clamp technique. 2. Membrane potentials showed prominent peaks at -35 mV and -70 mV. Membrane potentials of LPS-treated cells alternated between the two values. This may be due to a negative slope region of the I-V relation resulting in two zero current potentials. 3. From a holding potential of -70 mV, hyperpolarizing steps evoked an inwardly rectifying current both in proliferating and in LPS-treated cells, while depolarizing steps below -50 mV evoked an outwardly rectifying current only in LPS-treated microglia. The currents were K+ selective, as indicated by their reversal potential of approximately 0 mV in symmetric K+ concentrations (150 mM both intra- and extracellularly) and the reversal potential of the outward tail currents of approximately -90 mV at a normal extracellular K+ concentration (4.5 mM). 4. The activation of the outward current could be fitted by Hodgkin-Huxley-type n4 kinetics. The time constant of activation depended on voltage. 5. The inactivation of the inward and outward currents could be fitted by a single exponential. The time constant of the inward current inactivation was dependent on voltage, whereas the time constant of the outward current inactivation was virtually independent of voltage, except near the threshold of activation. Recovery of the outward from inactivation was slow and could be fitted by two exponentials. Responses to depolarizing steps were stable at 0.125 Hz, but greatly decreased from the first to the second pulse at 1 Hz. 6. The inactivation of the inward, but not of the outward, current disappeared in a low Na(+)-containing medium (5 mM). The inward current was selectively inhibited by extracellular Cs+ and Ba2+. The outward current was selectively inhibited by Cd2+, 4-aminopyridine and charybdotoxin. Replacement of intracellular K+ by an equimolar concentration of Cs+, and the extracellular application of tetraethylammonium and quinine inhibited both currents. 7. An increase of extracellular Ca2+ from 2 to 20 mM resulted in outwardly rectifying K+ channels activating at more positive potentials. Omission of Ca2+ from the extracellular medium had the opposite effect. When the intracellular free Ca2+ was increased from 0.01 to 1 microM, the outward current amplitudes were depressed. The Ca2+ ionophore A23187 had a similar effect. 8. LPS-treated microglial cells possess inwardly and outwardly rectifying K+ channels. The physiological and pharmacological characteristics of these two channel populations are markedly different.(ABSTRACT TRUNCATED AT 400 WORDS) 相似文献
9.
Postnatal rat nigrostriatal dopaminergic neurons exhibit five types of potassium conductances 总被引:3,自引:0,他引:3
1. We have investigated the electrical properties of neurons acutely dissociated from the substantia nigra zona compacta (SNZC) of the postnatal rat with whole cell patch-clamp recordings. Retrogradely labeled nigrostriatal neurons were identified with the use of rhodamine-labeled fluorescent latex microspheres. Over 90% of the rhodamine-labeled neurons in the SNZC demonstrated formaldehyde/glutaraldehyde-induced catecholamine fluorescence, indicating that they were dopaminergic (DA) neurons. 2. DA neurons had 15-20 microns ovoid or fusiform-shaped cell bodies with 2-3 thick proximal processes. Labeled neurons generated spontaneous action-potential activity in both regular and irregular patterns. These cells exhibited input resistances of 300-600 M omega and action-potential amplitudes of 60-80 mV. Locally applied dopamine inhibited the spontaneous activity of these neurons by hyperpolarizing the cells. 3. Outward currents were examined with voltage-clamp recordings using a tetrodotoxin (TTX)-containing medium. In all DA cells, depolarizing voltage commands activated several components of outward current depending on the holding potential of the cell. When cells were held at -40 mV (or more positive), voltage steps activated a sustained outward current. If the membrane potential was held more negative than -50 mV, a rapidly activating and inactivating component of outward current response could also be detected. 4. From a hyperpolarized holding potential (-90 mV) the transient outward current activated with depolarizing commands to -55 mV, peaking within 5 ms. The current inactivated with a monoexponential time constant of 53 +/- 4 (SE) ms. At more positive holding potentials (-40 mV) the steady-state inactivation of the current could be removed by applying a conditioning hyperpolarizing prepulse. In response to a fixed depolarizing voltage step, half-maximal inactivation occurred at about -65 mV. The transient current was blocked by 4-aminopyridine (4-AP). 5. The sustained outward currents were isolated by holding the cells at -40 mV. Two components of sustained outward current were distinguished by their sensitivity to the calcium channel blockers Co2+ (5 mM) and/or Cd2+ (200 microM). The current remaining in the presence of Co2+/Cd2+ was activated by depolarizing voltage commands more positive than -40 mV.(ABSTRACT TRUNCATED AT 400 WORDS) 相似文献
10.
Calcium channel currents in isolated guinea-pig ventricular cells superfused with Ca-free EGTA solution 总被引:1,自引:0,他引:1
Changes in membrane currents seen in Ca-free, EGTA (1 mM)-containing Tyrode solution (EGTA Tyrode), were studied in isolated guinea-pig ventricular cells, under the voltage clamp performed with a "G omega seal" patch electrode. Application of the EGTA Tyrode (calculated [Ca]0 = 1.3 X 10(-9) M) first eliminated the usual calcium current, but induced an extra inward current within 2 min. The reversal potential of this current, as judged by the direction of the current change, was about +25 mV (without correction of a liquid junction potential of -12 mV), but above this voltage a decaying outward current was observed. The decay of these inward and outward currents during depolarization was slow, but a large, nearly time-independent component was evident. These currents, regardless of their polarity and time course, were reduced by application of verapamil (10(-5) M) and Mg (5 mM), and were inactivated by pre-depolarizations. In Na-free EGTA Tyrode, the inward current disappeared but the outward current persisted at high voltages. These results suggest that in ventricular cells, reduction of external Ca concentrations to a nanomolar range induces a Ca channel current composed of an inward current carried by Na, and an outward current, presumably carried by K ions. Because of the persistence of the apparently non-inactivating Ca channel current, the net membrane current evoked at voltages around 0 mV remained close to zero, or even inward, after the decay of the time-dependent component, which was completed within a few hundreds ms. This characteristic I-V relation was considered to be linked to the development of the long-lasting action potentials, with a plateau maintained at around 0 mV, in EGTA Tyrode. 相似文献
11.
We measured and compared Na-Ca exchanger current (INa-Ca) from rabbit isolated ventricular and atrioventricular (AV) nodal myocytes, using action potential (AP) and ramp voltage commands. Whole cell patch-clamp recordings were made at 35-37 degrees C; INa-Ca was measured as 5 mM nickel (Ni)- sensitive current with major interfering voltage and calcium-activated currents blocked. In ventricular cells a 2-s descending ramp elicited INa-Ca showing outward rectification and a reversal potential (Erev) of -13.1 +/- 1. 2 mV (n = 12; mean +/- SEM). With a ventricular AP as the voltage command, the profile of INa-Ca followed the applied waveform closely. The current-voltage relation during AP repolarization was almost linear and showed an Erev of -38.3 +/- 5.3 mV (n = 6). As INa-Ca depended on the applied voltage waveform, comparisons between the two cell types utilized the same command waveform (a series of AV nodal APs). In ventricular myocytes this elicited INa-Ca that reversed near -38 mV and was inwardly directed during the pacemaker potential. This command was also applied to AV node cells; mean INa-Ca density at all voltages encompassed by the AP (-70 to +30 mV) did not differ significantly from that in ventricular myocytes (P > 0.05, ANOVA). This finding was confirmed using brief (250 ms) voltage ramp protocols (P > 0.1 ANOVA). These data represent the first direct measurements of AV nodal INa-Ca and suggest that the exchanger may be functionally expressed to similar levels in the two cell types. They may also suggest a possible role for INa-Ca during the pacemaker potential in AV node as inward INa-Ca was observed over the pacemaker potential range even with bulk internal Ca buffered to a low level. 相似文献
12.
Na(+) currents were studied by whole cell patch clamp of chalice-shaped afferent terminals attached to type I hair cells isolated from the gerbil semicircular canal and utricle. Outward K(+) currents were blocked with intracellular Cs(+) or with extracellularly applied 20 microM linopirdine and 2.5 mM 4-aminopyridine (4-AP). With K(+) currents blocked, inward currents activated and inactivated rapidly, had a maximum mean peak amplitude of 0.92 +/- 0.60 (SD) nA (n = 24), and activated positive to -60 mV from holding potentials of -70 mV and more negative. The transient inward currents were blocked almost completely by 100 nM TTX, confirming their identity as Na(+) currents. Half-inactivation of Na(+) currents occurred at -82.6 +/- 0.9 mV, with a slope factor of 9.2 +/- 0.8 (n = 7) at room temperature. In current clamp, large overshooting action potential-like events were observed only after prior hyperpolarizing current injections. However, spontaneous currents consistent with quantal release from the hair cell were observed at holding potentials close to the zero-current potential. This is the first report of ionic conductances in calyx terminals postsynaptic to type I hair cells in the mammalian vestibular system. 相似文献
13.
Outward currents through the inwardly rectifying potassium channel of guinea-pig ventricular cells 总被引:2,自引:0,他引:2
Currents through the inwardly rectifying K channel were studied under whole-cell clamp of collagenase-treated single ventricular cells of guinea-pigs. The inwardly rectifying K channel was fully activated by hyperpolarizing the membrane from the equilibrium potential for K+ (EK) by 30-40 mV. Following depolarization above EK, a decaying outward current was elicited. Prolongation of the hyperpolarizing prepulse increased the amplitude of the decaying outward current, with a time course similar to the increase of the inward current during the prepulse. Time-dependent changes in both outward and inward currents could be fitted with a single exponential function and were attributed to deactivation and activation of the inwardly rectifying K channel. The instantaneous current-voltage relation was almost linear, indicating that the conductance of the channel is ohmic and that the rectification of the steady-state current was due to the kinetic properties of the inwardly rectifying K channel. The activation kinetics of the channel was measured at different concentrations of K+ in both the external and internal solutions. The time constant and the steady-state activation were not a function of the absolute membrane potential value, but were dependent on the driving force. 相似文献
14.
Alterations in the state of excitability of midbrain dopamine (DA) neurons from the ventral tegmental area (VTA) may underlie changes in the synaptic plasticity of the mesocorticolimbic system. Here, we investigated norepinephrine's (NE) regulation of VTA DA cell excitability by modulation of the hyperpolarization-activated cation current, Ih, with whole cell recordings in rat brain slices. Current clamp recordings show that NE (40 microM) hyperpolarizes spontaneously firing VTA DA cells (11.23+/-4 mV; n=8). In a voltage clamp, NE (40 microM) induces an outward current (100+/-24 pA; n=8) at -60 mV that reverses at about the Nernst potential for potassium (-106 mV). In addition, NE (40 microM) increases the membrane cord conductance (179+/-42%; n=10) and reduces Ih amplitude (68+/-3% of control at -120 mV; n=10). The noradrenergic alpha-1 antagonist prazosin (40 microM; n=5) or the alpha-2 antagonist yohimbine (40 microM; n=5) did not block NE effects. All NE-evoked events were blocked by the D2 antagonists sulpiride (1 microM) and eticlopride (100 nM) and no significant reduction of Ih took place in the presence of the potassium channel blocker BaCl2 (300 microM). Therefore, it is concluded that NE inhibition of Ih was due to an increase in membrane conductance by a nonspecific activation of D2 receptors that induce an outward potassium current and is not a result of a second messenger system acting on h-channels. The results also suggest that Ih channels are mainly located at dendrites of VTA DA cells and, thus, their inhibition may facilitate the transition from single-spike firing to burst firing and vice versa. 相似文献
15.
1. Bipolar cells were isolated from adult rat retinas after enzymatic and mechanical treatment. The cells could be unequivocally identified from their morphology because of high retention of their axon and dendritic processes after isolation. 2. Protein kinase C (PKC) immunoreactivity performed on sections of the rat retina labeled rod bipolar cells and a few amacrine cells. Virtually all bipolar cells in the dissociates expressed PKC immunoreactivity and were, therefore, rod bipolar cells. 3. Rod bipolar cells were examined with the tight-seal whole-cell and excised-patch recording techniques. Resting potentials of the isolated cells recorded under current-clamp conditions showed a broad unimodal distribution around -37 mV. 4. Membrane depolarization from a holding potential of -90 mV resulted in an outward current. A fast sodium inward current was not observed. Membrane hyperpolarization from a holding potential of -40 mV activated an inwardly rectifying current. 5. gamma-Aminobutyric acid (GABA) and glycine, the putative retinal neurotransmitters that mediate the bipolar cells' receptive field surround in vivo, activated chloride conductances in almost all isolated bipolar cells. GABA- and glycine-evoked currents were both desensitizing and could be antagonized by the classical blockers bicuculline, picrotoxin, and strychnine, respectively. 6. Pressure application of the drugs from fine microcapillaries to various parts of the isolated cells suggests a high GABA sensitivity at the axonal endings compared with either the somatic or dendritic region. A similar distribution was not found for glycine. On the contrary, glycine-induced single-channel events with main conductances of 52 and 34 pS were recorded from membrane patches excised from the cells' somata. 7. Conductances induced by glutamate and several excitatory amino acid agonists were observed in a number of the cells. Application of the glutamate agonist 2-amino-4-phosphonobutyric acid (APB) induced an inward current at negative holding potentials associated with the opening of ion channels. In only 5 of 93 cells, APB closed ion channels, leading to a decrease in membrane conductance. 相似文献
16.
R. D. Smith D. A. Eisner Susan Wray 《Pflügers Archiv : European journal of physiology》1998,435(4):518-522
Guinea-pig ureteric smooth muscle is unusual in that intracellular acidification increases and alkalinization decreases force
production. To help elucidate the mechanism underlying these effects on force we have investigated the effects of changing
intracellular pH on both calcium and potassium currents in single cells isolated from the guinea-pig ureter to determine their
possible role in force development. Depolarization to +40 mV resulted in a fast transient outward current which was inhibited
by 4-aminopyridine but not tetraethylammonium. Intracellular alkalinization (20 mM trimethylamine) increased this current
to 179 ± 24% of the control and resulted in the development of a slowly activating large outward current which was inhibited
by tetraethylammonium and washout. Acidification (40 mM sodium butyrate) decreased the fast transient outward current to 58 ± 3%
of the control and did not produce a slowly activating current. When potassium was replaced by caesium in the pipette solution,
depolarization to 0 mV resulted in an inward calcium current which was abolished by nifedipine. Intracellular alkalinization
increased this current to 126 ± 11% of the control whereas acidification had the opposite effect, decreasing it to 55 ± 10%.
Furthermore, current-clamp experiments showed that intracellular alkalinization inhibited the amplitude of the action potential,
therefore decreasing excitability of the cell. From our results, we suggest that the predominant effects of intracellular
pH on force production in the guinea-pig ureter are mediated via the modulation of outward potassium currents (thereby reducing
excitability of the tissue) rather than the effects on the inward calcium current.
Received: 31 July 1997 / Received after revision: 27 October 1997 / Accepted: 28 October 1997 相似文献
17.
Ionic currents in crustacean neurosecretory cells. 总被引:2,自引:0,他引:2
1. The patterns of electrical activity and membrane characteristics of a population of neurosecretory-cell somata in the X-organ of the crayfish were investigated with microelectrodes and whole-cell, voltage-clamp techniques. Some neurons (56%) were silent but could be excited by intracellular current injection: other cells showed spontaneous tonic activity (35%), and some had spontaneous bursting activity (9%). The spiking activity was abolished by tetrodotoxin (TTX) exposure and by severing the axon near the cell body. After axotomy, only a small, slow, regenerative depolarization remained that could be blocked by Cd2+. 2. Under voltage clamp the steady-state I-V curve in low [Ca2+]i (9 X 10(-9) M) showed a slope conductance of 16.7 +/- 3.9 (SD) nS (n = 10) at -50 mV and zero current potential of -50.1 +/- 7.7 mV. In current-clamp mode these neurons were either silent or fired tonically. With high [Ca2+]i (1.7 X 10(-6) M) both the slope conductance and inward and outward currents were reduced. In some neurons high [Ca2+]i reveals a negative slope resistance in the range of -46 to -41 mV. It could be supressed by removing [Na+]o, but it was TTX insensitive. These are the neurons that under current clamp showed bursting activity. 3. The main inward current in cell somata was a Ca2+ current of 2 +/- 0.6 nA (n = 18), activated at -40 mV and peaking at 20 mV. It showed relaxation with prolonged pulses. No Na(+)-dependent, TTX-sensitive inward currents were recorded with short (100-ms) pulses in axotomized neurons. 4. Two outward currents could be distinguished.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
18.
Crustacean cardiac ganglion neuronal somata, although incapable of generating action potentials, produce regenerative, slow (greater than 200 ms) depolarizing potentials reaching -20 mV (from -50 mV) in response to depolarizing stimuli. These potentials initiate a burst of action potentials in the axon and are thus termed driver potentials. The somata of the anterior-most neurons (cells 1 or 2) were isolated by ligaturing for study of their membrane currents with a two-electrode voltage clamp. Inward current is attributed to Ca2+ by reason of dependence of driver potential amplitude on [Ca2+]0, independence of [Na+]0, resistance to tetrodotoxin, and inhibition by Cd (0.2 mM) and Mn (4 mM). Ca-mediated current (ICa) is present at -40 mV. It is optimally activated by a holding potential (Vh) of -50 to -60 mV and by clamps (command potential, Vc) to -10 mV. Time to peak (10-30 ms) and amplitude are strongly voltage dependent. Maximum tail-current amplitudes observed at -70 to -85 mV are ca. 100 nA. Inward tail peaks may not be resolved by our clamp (settling time, 2 ms). Tails relax with a time constant (tau) of approximately equal to 12 ms (at -70 to -85 mV). ICa exhibits inactivation in double pulse regimes. Recovery has a tau of approximately equal to 0.7 s. Tail current analyses indicate an exponential decline (tau approximately equal to 23 ms at -20 mV) toward a maintained amplitude of inward current tails. Analysis of outward currents indicates the presence of three conductance mechanisms having voltage dependences, time courses, and pharmacology similar to those of early outward current (IA), delayed outward current (IK), and outward current (IC) of molluscan neurons. Analysis of tail currents indicates a reversal potential for each of these near -75 mV, indicating that they are K currents. Early outward current, IA, shows a peak at 5 ms followed by rapid decline. Response to a second clamp given within 0.4 s is reduced; recovery is exponential, with a tau of approximately equal to 200 ms (at Vh = -50 mV). The amplitude of IA tested at 0 mV shows activation or deactivation by subthreshold shifts of Vh. The extent and rate of these changes shows voltage dependence (tau approximately equal to 100-500 ms for subthreshold prepulses). At the normal cell resting potential of -50 mV the amplitude of IA is 25% of that tested from -80 mV.(ABSTRACT TRUNCATED AT 400 WORDS) 相似文献
19.
Duffy SM Lawley WJ Kaur D Yang W Bradding P 《The Journal of allergy and clinical immunology》2003,112(5):965-972
BACKGROUND: Human lung mast cells (HLMCs) and the human mast cell line HMC-1 express a strongly outwardly rectifying Cl- current characteristic of that carried by the voltage-dependent Cl- channel ClC-5. A similar but distinct current has been implicated in the control of cell proliferation in astrocytes. OBJECTIVE: In this study, we have examined the effects of the Cl- channel blocker tamoxifen on ion channel activity and cell proliferation in both HMC-1 and HLMCs. METHODS: We used the whole-cell patch-clamp technique to characterize macroscopic ion currents in mast cells before and after addition of tamoxifen. HMC-1 proliferation was assessed by incorporation of tritiated thymidine, HLMC proliferation was determined by counting cells in long-term culture, and cell viability was assessed by annexin V binding and propidium iodide uptake. RESULTS: In HMC-1, tamoxifen reduced the outward Cl- current at +130 mV by 73% +/- 9% at a concentration of 3 micromol/L and simultaneously opened a novel inwardly rectifying nonselective cation current with a mean inward current of 153 +/- 18 pA at -130 mV. Tamoxifen produced a dose-dependent inhibition of HMC-1 proliferation (90.3% +/- 4.0% inhibition at 30 micromol/L) without altering cell viability. Tamoxifen inhibited the outward ClC-5-like current in HLMCs, did not open an inward current, and produced a dose-dependent inhibition of HLMC proliferation in long-term culture. CONCLUSION: Tamoxifen inhibits HMC proliferation, possibly through ion channel modulation. This suggests that tamoxifen might be useful in the treatment of mast-cell-mediated diseases, including mastocytosis, asthma, and pulmonary fibrosis. 相似文献
20.
Single channel analysis of the inward rectifier K current in the rabbit ventricular cells 总被引:5,自引:0,他引:5
The inward rectifier K channel in rabbit ventricular cells was studied by the patch-clamp method. Single channel currents were recorded in giga-sealed cell-attached patches with 150 mM K+ in the pipette. The slope conductance in the membrane potential range from -140 to -40 mV was 46.6 +/- 6.7 pS (mean +/- S.D., n = 16), and was reduced by decreasing [K+] in the pipette (20 or 50 mM). The channel was blocked by an application of Cs+ or Ba2+ (0.04-1 mM) in the pipette. Outwardly directed current, recorded with 50 mM K+ in the pipette, revealed the inward rectification of the single channel current. The probability of the channel being open was 0.33 +/- 0.05 (n = 10) at the resting potential (RP=-81.7 +/- 1.7 mV, n = 16) with 150 mM K+ in the pipette, and it decreased with hyperpolarization. The mean open time of the channel was 178 +/- 25 msec (n = 6) at RP. The closed time of the channel seemed to have two exponential components, with time constants of 11.0 +/- 2.0 msec and 1.92 +/- 0.52 sec (n = 6) at RP. The slower time constant was increased with hyperpolarization. The averaged patch current recorded upon hyperpolarizing pulses demonstrated a time-dependent current decay as expected from the single channel kinetics. The results indicated that the inward rectifier K+ current has time- and voltage-dependent kinetics. 相似文献