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1.
Taddese A  Bean BP 《Neuron》2002,33(4):587-600
A role for "persistent," subthreshold, TTX-sensitive sodium current in driving the pacemaking of many central neurons has been proposed, but this has been impossible to test pharmacologically. Using isolated tuberomammillary neurons, we assessed the role of subthreshold sodium current in pacemaking by performing voltage-clamp experiments using a cell's own pacemaking cycle as voltage command. TTX-sensitive sodium current flows throughout the pacemaking cycle, even at voltages as negative as -70 mV, and this current is sufficient to drive spontaneous firing. When sodium channels underlying transient current were driven into slow inactivation by rapid stimulation, persistent current decreased in parallel, suggesting that persistent sodium current originates from subthreshold gating of the same sodium channels that underlie the phasic sodium current. This behavior of sodium channels may endow all neurons with an intrinsic propensity for rhythmic, spontaneous firing.  相似文献   

2.
Dopaminergic local circuit neurons in the retina (DA cells) show robust, spontaneous, tetrodotoxin-sensitive pacemaking. To investigate the mechanism underlying this behavior, we characterized the sodium current and a subset of the potassium currents in the cells in voltage-clamp experiments. We found that there is a persistent component of the sodium current in DA cells which activates at more depolarized potentials than the transient component of the current. The transient component was completely inactivated at -50 mV, but DA cells remained able to fire spontaneous action potentials when potassium channels were partially blocked and the membrane potential remained above -40 mV. Based on these electrophysiological data, we developed a reduced computer model that reproduced the major features of DA cells. In simulations at the physiological resting potential, the persistent component of the sodium current was both necessary and sufficient to account for spontaneous activity, and the major contribution of the transient component of the sodium current was to initiate the depolarization of the model cell during the interspike interval. When tonic inhibition was simulated by lowering the input impedance of the model cell, the transient component played a larger role.  相似文献   

3.
Midbrain dopaminergic neurons are endowed with endogenous slow pacemaking properties. In recent years, many different groups have studied the basis for this phenomenon, often with conflicting conclusions. In particular, the role of a slowly-inactivating L-type calcium channel in the depolarizing phase between spikes is controversial, and the analysis of slow oscillatory potential (SOP) recordings during the blockade of sodium channels has led to conflicting conclusions. Based on a minimal model of a dopaminergic neuron, our analysis suggests that the same experimental protocol may lead to drastically different observations in almost identical neurons. For example, complete L-type calcium channel blockade eliminates spontaneous firing or has almost no effect in two neurons differing by less than 1% in their maximal sodium conductance. The same prediction can be reproduced in a state of the art detailed model of a dopaminergic neuron. Some of these predictions are confirmed experimentally using single-cell recordings in brain slices. Our minimal model exhibits SOPs when sodium channels are blocked, these SOPs being uncorrelated with the spiking activity, as has been shown experimentally. We also show that block of a specific conductance (in this case, the SK conductance) can have a different effect on these two oscillatory behaviors (pacemaking and SOPs), despite the fact that they have the same initiating mechanism. These results highlight the fact that computational approaches, besides their well known confirmatory and predictive interests in neurophysiology, may also be useful to resolve apparent discrepancies between experimental results.  相似文献   

4.
Currents generated by depolarizing voltage pulses were recorded in neurons from the pyramidal cell layer of the CA1 region of rat or guinea pig hippocampus with single electrode voltage-clamp or tight-seal whole-cell voltage-clamp techniques. In neurons in situ in slices, and in dissociated neurons, subtraction of currents generated by identical depolarizing voltage pulses before and after exposure to tetrodotoxin revealed a small, persistent current after the transient current. These currents could also be recorded directly in dissociated neurons in which other ionic currents were effectively suppressed. It was concluded that the persistent current was carried by sodium ions because it was blocked by TTX, decreased in amplitude when extracellular sodium concentration was reduced, and was not blocked by cadmium. The amplitude of the persistent sodium current varied with clamp potential, being detectable at potentials as negative as -70 mV and reaching a maximum at approximately -40 mV. The maximum amplitude at -40 mV in 21 cells in slices was -0.34 +/- 0.05 nA (mean +/- 1 SEM) and -0.21 +/- 0.05 nA in 10 dissociated neurons. Persistent sodium conductance increased sigmoidally with a potential between -70 and -30 mV and could be fitted with the Boltzmann equation, g = gmax/(1 + exp[(V' - V)/k)]). The average gmax was 7.8 +/- 1.1 nS in the 21 neurons in slices and 4.4 +/- 1.6 nS in the 10 dissociated cells that had lost their processes indicating that the channels responsible are probably most densely aggregated on or close to the soma. The half-maximum conductance occurred close to -50 mV, both in neurons in slices and in dissociated neurons, and the slope factor (k) was 5-9 mV. The persistent sodium current was much more resistant to inactivation by depolarization than the transient current and could be recorded at greater than 50% of its normal amplitude when the transient current was completely inactivated. Because the persistent sodium current activates at potentials close to the resting membrane potential and is very resistant to inactivation, it probably plays an important role in the repetitive firing of action potentials caused by prolonged depolarizations such as those that occur during barrages of synaptic inputs into these cells.  相似文献   

5.
Properties of the calcium-activated chloride current in heart   总被引:12,自引:0,他引:12       下载免费PDF全文
We used the whole cell patch clamp technique to study transient outward currents of single rabbit atrial cells. A large transient current, IA, was blocked by 4-aminopyridine (4AP) and/or by depolarized holding potentials. After block of IA, a smaller transient current remained. It was completely blocked by nisoldipine, cadmium, ryanodine, or caffeine, which indicates that all of the 4AP-resistant current is activated by the calcium transient that causes contraction. Neither calcium-activated potassium current nor calcium-activated nonspecific cation current appeared to contribute to the 4AP-resistant transient current. The transient current disappeared when ECl was made equal to the pulse potential; it was present in potassium-free internal and external solutions. It was blocked by the anion transport blockers SITS and DIDS, and the reversal potential of instantaneous current-voltage relations varied with extracellular chloride as predicted for a chloride-selective conductance. We concluded that the 4AP-resistant transient outward current of atrial cells is produced by a calcium-activated chloride current like the current ICl(Ca) of ventricular cells (1991. Circulation Research. 68:424-437). ICl(Ca) in atrial cells demonstrated outward rectification, even when intracellular chloride concentration was higher than extracellular. When ICa was inactivated or allowed to recover from inactivation, amplitudes of ICl(Ca) and ICa were closely correlated. The results were consistent with the view that ICl(Ca) does not undergo independent inactivation. Tentatively, we propose that ICl(Ca) is transient because it is activated by an intracellular calcium transient. Lowering extracellular sodium increased the peak outward transient current. The current was insensitive to the choice of sodium substitute. Because a recently identified time-independent, adrenergically activated chloride current in heart is reduced in low sodium, these data suggest that the two chloride currents are produced by different populations of channels.  相似文献   

6.
The interstitial cells of Cajal (ICC) are responsible for producing pacemaking signals that stimulate rhythmic contractions in the gastro-intestinal system. The pacemaking signals are generated by membrane depolarizations, which are in turn linked to the integrated transport of calcium between the endoplasmic reticulum (ER), through inositol-trisphosphate receptor (IP3R) release, and mitochondria, through the uniporter. A non-specific cation channel (NSCC) is associated with the membrane depolarizations, and is inhibited by intracellular calcium. One theory proposes that the integrated calcium transport occurs within specific regions of the ICC called “pacemaker units,” and results in localized calcium concentration reductions within these units, which in turn activate the NSCC and depolarize the membrane. We have constructed a model of the spatio-temporal calcium dynamics within an ICC pacemaker unit to determine under what conditions the local calcium concentrations may reduce below baseline. We obtain reductions of calcium concentrations below baseline but only under certain conditions. Without strong and persistent stimulation of the IP3R, reductions of calcium below baseline occur only with a non-physiological, time-dependent uniporter. Alternatively, sufficient IP3R release leads to reductions of calcium below baseline, due to depletion of the ER calcium store over the time scale of seconds, although these reductions require strong mitochondrial and ER calcium uptake.  相似文献   

7.
The objective of these studies was to define the roles of calcium and sodium in uterine smooth muscle excitation. The double sucrose-gap technique was used for current-clamp and voltage-clamp experiments. It was shown that neither sodium nor calcium alone is capable of supporting excitation in estrogen-dominated uterine smooth muscle. Calcium dependence was explained in part by increased membrane "leakage" current in calcium-free solution and calcium control of the voltage dependence of the early transient conductance. High concentrations of TTX did not affect the magnitude of the peak transient current while La+++, Mn++, and Co++ greatly reduced or abolished it and decreased the steady-state current. From these and other data it was concluded that the regenerative mechanism in uterine smooth muscle has the functional characteristics of a single transient conductance channel whose activation requires the presence of both sodium and calcium. Insensitivity to TTX indicates that the molecular structure of the channel is unlike that in certain sodium-dependent systems, while the effects of La+++, Mn++, Co++, and Ca++ reveal a similar dependence of conductances on extracellular polyvalent cations.  相似文献   

8.
A cardiac biological pacemaker (BP) has been created by suppression of the inward rectifier K(+) current (I(K1)) or overexpression of the hyperpolarization-activated current (I(h)). We theoretically investigated the effects of incorporating I(h), T-type Ca(2+) current (I(Ca,T)), sustained inward current (I(st)), and/or low-voltage-activated L-type Ca(2+) channel current (I(Ca,LD)) on 1) creation of BP cells, 2) robustness of BP activity to electrotonic loads of nonpacemaking (NP) cells, and 3) BP cell ability to drive NP cells. We used a single-cell model for human ventricular myocytes (HVMs) and also coupled-cell models composed of BP and NP cells. Bifurcation structures of the model cells were explored during changes in conductance of the currents and gap junction. Incorporating the pacemaker currents did not yield BP activity in HVM with normal I(K1) but increased the critical I(K1) conductance for BP activity to emerge. Expressing I(h) appeared to be most helpful in facilitating creation of BP cells via I(K1) suppression. In the coupled-cell model, I(st) significantly enlarged the gap conductance (G(C)) region where stable BP cell pacemaking and NP cell driving occur, reducing the number of BP cells required for robust pacemaking and driving. In contrast, I(h) enlarged the G(C) region of pacemaking and driving only when I(K1) of the NP cell was relatively low. I(Ca,T) or I(Ca,LD) exerted effects similar to those of I(st) but caused shrinkage or irregularity of BP oscillations. These findings suggest that expressing I(st) most effectively improves the structural stability of BPs to electrotonic loads and the BP ability to drive the ventricle.  相似文献   

9.
Summary Voltage-current curves for the isolated midgut of the tobacco hornworm were determined by transient and steady voltage clamping over the range of 200 to –200 mV. Over this range the transient method yields a linear relation while the steady method usually yields a curve consisting of two lines of differing slope which intersect at zero voltage. The difference between the results of the methods is due to a slow decline in total conductance which accompanies steady voltage clamping.Holding the midgut at short circuit increases the total conductance of the tissue in a manner consistent with increasing shunt conductance; this effect was seen in both diet-reared and leaf-reared animals.When potassium transport is inhibited by substitution of choline or sodium for potassium in bathing solution the total conductance decreases and the voltage-current curve intersects the normal curve in the hyperpolarizing region. Applying a simple equivalent circuit analysis to the results from partial or total potassium replacement suggests that the electromotive force of the potassium transport system is of the order of 140–190 mV. The conductance decrease during inhibition of potassium transport by transient anoxia is of similar magnitude, suggesting that a major effect of metabolic inhibition is to decrease the active conductance of the potassium transport pathway.  相似文献   

10.
Mutations in genes encoding neuronal voltage-gated sodium channel subunits have been linked to inherited forms of epilepsy. The majority of mutations (>100) associated with generalized epilepsy with febrile seizures plus (GEFS+) and severe myoclonic epilepsy of infancy (SMEI) occur in SCN1A encoding the NaV1.1 neuronal sodium channel alpha-subunit. Previous studies demonstrated functional heterogeneity among mutant SCN1A channels, revealing a complex relationship between clinical and biophysical phenotypes. To further understand the mechanisms responsible for mutant SCN1A behavior, we performed a comprehensive analysis of the single-channel properties of heterologously expressed recombinant WT-SCN1A channels. Based on these data, we then determined the mechanisms for dysfunction of two GEFS+-associated mutations (R1648H, R1657C) both affecting the S4 segment of domain 4. WT-SCN1A has a slope conductance (17 pS) similar to channels found in native mammalian neurons. The mean open time is approximately 0.3 ms in the -30 to -10 mV range. The R1648H mutant, previously shown to display persistent sodium current in whole-cell recordings, exhibited similar slope conductance but had an increased probability of late reopening and a subfraction of channels with prolonged open times. We did not observe bursting behavior and found no evidence for a gating mode shift to explain the increased persistent current caused by R1648H. Cells expressing R1657C exhibited conductance, open probability, mean open time, and latency to first opening similar to WT channels but reduced whole-cell current density, suggesting decreased number of functional channels at the plasma membrane. In summary, our findings define single-channel properties for WT-SCN1A, detail the functional phenotypes for two human epilepsy-associated sodium channel mutants, and clarify the mechanism for increased persistent sodium current induced by the R1648H allele.  相似文献   

11.
This paper deals with the physical interpretation of existing mathematical models which describe the transient sodium conductance changes in excitable membranes. It is shown that there are clear limitations to the specificity of inferences which may be drawn about physical mechanism from the behavior of abstract models. Within these limitations, it is shown that a pronounced inactivation shift is not necessarily evidence for coupling between the events responsible for the rise and inactivation of the sodium conductance, but that the inactivation shift may be associated with an event whose rate explicitly depends on the rate of continuous voltage change or magnitude of instantaneous voltage change.  相似文献   

12.
A N Zubov  K V Sal'nikov 《Tsitologiia》1986,28(12):1336-1344
Adding of 5% bovine serum to internally perfused voltage-clamped serum deprived neuroblastoma cells rapidly stimulates transient sodium current. This stimulating effect is mainly due to the increase in the peak sodium conductance by almost 24 per cent, on the average. Besides that a modifying effect was observed resulting in the 6 mV shift of the sodium peak conductance curve towards more negative potentials and in the 5 mV shift of steady inactivation curve towards more positive ones. The sign of the latter shift was changed to the opposite under the action of serum thermally pretreated at 100 degrees C. This procedure led also to more than two fold lowering of the stimulating effect. Experiments with serum deprivation demonstrate different degrees of reversibility of the serum effects, the most reversible being the inactivation curve shift. EGF, insulin, dexamethasone, transferrin, ATP, serotonin and their combinations in physiological concentrations failed to give the typical whole serum effects. The serum is supposed to contain at least two active components of unknown nature, one of which being thermoresistant.  相似文献   

13.
To elucidate the regional differences in sinoatrial node pacemaking mechanisms, we investigated 1), bifurcation structures during current blocks or hyperpolarization of the central and peripheral cells, 2), ionic bases of regional differences in bifurcation structures, and 3), the role of Na+ channel current (INa) in peripheral cell pacemaking. Bifurcation analyses were performed for mathematical models of the rabbit sinoatrial node central and peripheral cells; equilibrium points, periodic orbits, and their stability were determined as functions of parameters. Structural stability against applications of acetylcholine or electrotonic modulations of the atrium was also evaluated. Blocking L-type Ca2+ channel current (ICa,L) stabilized equilibrium points and abolished pacemaking in both the center and periphery. Critical acetylcholine concentration and gap junction conductance for pacemaker cessation were higher in the periphery than in the center, being dramatically reduced by blocking INa. Under hyperpolarized conditions, blocking INa, but not eliminating ICa,L, abolished peripheral cell pacemaking. These results suggest that 1), ICa,L is responsible for basal pacemaking in both the central and peripheral cells, 2), the peripheral cell is more robust in withstanding hyperpolarizing loads than the central cell, 3), INa improves the structural stability to hyperpolarizing loads, and 4), INa-dependent pacemaking is possible in hyperpolarized peripheral cells.  相似文献   

14.
We investigate spike initiation and propagation in a model axon that has a slow regenerative conductance as well as the usual Hodgkin-Huxley type sodium and potassium conductances. We study the role of slow conductance in producing repetitive firing, compute the dispersion relation for an axon with an additional slow conductance, and show that under appropriate conditions such an axon can produce a traveling zone of secondary spike initiation. This study illustrates some of the complex dynamics shown by excitable membranes with fast and slow conductances.  相似文献   

15.
The Hodgkin-Huxley equations for space-clamped squid axon (18 degrees C) have been modified to approximate voltage clamp data from repetitive-firing crustacean walking leg axons and activity in response to constant current stimulation has been computed. The m infinity and h infinity parameters of the sodium conductance system were shifted along the voltage axis in opposite directions so that their relative overlap was increased approximately 7 mV. Time constants tau m and tau h, were moved in a similar manner. Voltage-dependent parameters of delayed potassium conductance, n infinity and tau n, were shifted 4.3 mV in the positive direction and tau n was uniformly increased by a factor of 2. Leakage conductance and capacitance were unchanged. Repetitive activity of this modified circuit was qualitatively similar to that of the standard model. A fifth branch was added to the circuit representing a transient potassium conductance system present in the repetitive walking leg axons and in other repetitive neurons. This model, with various parameter choices, fired repetitively down to approximately 2 spikes/s and up to 350/s. The frequency vs. stimulus current plot could be fit well by a straight line over a decade of the low frequency range and the general appearance of the spike trains was similar to that of other repetitive neurons. Stimulus intensities were of the same order as those which produce repetitive activity in the standard Hodgkin-Huxley axon. The repetitive firing rate and first spike latency (utilization time) were found to be most strongly influenced by the inactivation time constant of the transient potassium conductance (tau b), the delayed potassium conductance (tau n), and the value of leakage conductance (gL). The model presents a mechanism by which stable low frequency discharge can be generated by millisecond-order membrane conductance changes.  相似文献   

16.
CLC Cl- channels are homodimers in which each subunit has a proper pore and a (fast) gate. An additional slow gate acts on both pores. A conserved glutamate (E166 in CLC-0) is a major determinant of gating in CLC-0 and is crucially involved in Cl-/H+ antiport of CLC-ec1, a CLC of known structure. We constructed tandem dimers with one wild-type (WT) and one mutant subunit (E166A or E166D) to show that these mutations of E166 specifically alter the fast gate of the pore to which they belong without effect on the fast gate of the neighboring pore. In addition both mutations activate the common slow gate. E166A pores have a large, voltage-independent open probability of the fast gate (popen), whereas popen of E166D pores is dramatically reduced. Similar to WT, popen of E166D was increased by lowering pHint. At negative voltages, E166D presents a persistent inward current that is blocked by p-chlorophenoxy-acetic acid (CPA) and increased at low pHext. The pHext dependence of the persistent current is analogous to a similar steady inward current in WT CLC-0. Surprisingly, however, the underlying unitary conductance of the persistent current in E166D is about an order of magnitude smaller than that of the transient deactivating inward Cl- current. Collectively, our data support the possibility that the mutated CLC-0 channel E166D can assume two distinct open states. Voltage-independent protonation of D166 from the outside favors a low conductance state, whereas protonation from the inside favors the high conductance state.  相似文献   

17.
Ion permeation through voltage-gated sodium channels is modulated by various drugs and toxins. The atomistic mechanisms of action of many toxins are poorly understood. A steroidal alkaloid batrachotoxin (BTX) causes persistent channel activation by inhibiting inactivation and shifting the voltage dependence of activation to more negative potentials. Traditionally, BTX is considered to bind at the channel-lipid interface and allosterically modulate the ion permeation. However, amino acid residues critical for BTX action are found in the inner helices of all four repeats, suggesting that BTX binds in the pore. In the octapeptide segment IFGSFFTL in IIIS6 of a cockroach sodium channel BgNa(V), besides Ser_3i15 and Leu_3i19, which correspond to known BTX-sensing residues of mammalian sodium channels, we found that Gly_3i14 and Phe_3i16 are critical for BTX action. Using these data along with published data as distance constraints, we docked BTX in the Kv1.2-based homology model of the open BgNa(V) channel. We arrived at a model in which BTX adopts a horseshoe conformation with the horseshoe plane normal to the pore axis. The BTX ammonium group is engaged in cation-π interactions with Phe_3i16 and BTX moieties interact with known BTX-sensing residues in all four repeats. Oxygen atoms at the horseshoe inner surface constitute a transient binding site for permeating cations, whereas the bulky BTX molecule would resist the pore closure, thus causing persistent channel activation. Our study reinforces the concept that steroidal sodium channel agonists bind in the inner pore of sodium channels and elaborates the atomistic mechanism of BTX action.  相似文献   

18.
Persistent use-dependent changes in the intrinsic neuronal excitability determine the long-term dynamics of the activity of these neurons. In synergy with the long-lasting modification of synaptic transmission, such changes in the excitability presumably contribute to the formation of a memory trace in the brain. Nevertheless, neither particular transmembrane ion conductances implicated in the intrinsic plasticity nor the mechanisms of regulation of such conductances have been identified in most neurons where this plasticity was observed. In our model study, we tried to determine those membrane conductances in cerebellar granule cells (GrCs) whose changes can result in a persistent increase in the input resistance and a decrease in the spike threshold observed after high-frequency stimulation of presynaptic neurons. For this purpose, published experimental results were simulated with the use of a slightly modified model of the electroresponsiveness of rat cerebellar GrCs. It was concluded that experimentally observed changes in the input resistance of the neuron, in the minimum current step needed to fire action potentials (APs), in the spike threshold, in the average spike frequency, and in the delay of the first spike may be caused only by changes in the background voltage-independent potassium conductance and persistent sodium conductance. Hyperpolarization-directed shifts in the activation and inactivation curves of fast sodium channels are also possible. The observed changes in the intrinsic excitability evoke the shift in the peak of the frequency-response curve in such a manner that it becomes close to the frequency of oscillations recorded in the cerebellar granular layer during realization of voluntary movements. Neirofiziologiya/Neurophysiology, Vol. 38, No. 2, pp. 119–130, March–April, 2006.  相似文献   

19.
The delayed outward current in snail neurones was separated into two components with different temperature sensitivity: (i) a persistent component and (ii) a transient (inactivating) component. The effect of cooling on the value of the transient current is strongly dependent upon the value of the conditioning potential. It was supposed that cooling causes a decrease in the negative surface potential in the vicinity of the potassium pathways and removes their inactivation. Simultaneously cooling depresses the potassium conductance. The effect on surface potential is more distinct with conditioning potentials at which a significant fraction of the transient outward current is inactivated. The effect of cooling on the transient component of the fast outward current was similar to that on the transient component of the delayed outward current.  相似文献   

20.
Cardiac pacemaking in the sinoatrial (SA) node and atrioventricular (AV) node is generated by an interplay of many ionic currents, one of which is the funny pacemaker current (If). To understand the functional role of If in two different pacemakers, comparative studies of spontaneous activity and expression of the HCN channel in mouse SA node and AV node were performed. The intrinsic cycle length (CL) is 179±2.7 ms (n=5) in SA node and 258±18.7 ms (n=5) in AV node. Blocking of If current by 1 μmol/L ZD7288 increased the CL to 258±18.7 ms (n=5) and 447±92.4 ms (n=5) in SA node and AV node, respectively. However, the major HCN channel, HCN4 expressed at low level in the AV node compared to the SA node. To clarify the discrepancy between the functional importance of If and expression level of HCN4 channel, a SA node cell model was used. Increasing the If conductance resulted in decreasing in the CL in the model, which explains the high pacemaking rate and high expression of HCN channel in the SA node. Resistance to the blocking of If in the SA node might result from compensating effects from other currents (especially voltage sensitive currents) involved in pacemaking. The computer simulation shows that the difference in the intrinsic CL could explain the difference in response to If blocking in these two cardiac nodes.  相似文献   

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