Sodium-activated Potassium Current Research Articles

Functional Coupling of Slack Channels and P2X3 Receptors Contributes to Neuropathic Pain Processing.

The sodium-activated potassium channel Slack (KNa1.1, Slo2.2, or Kcnt1) is highly expressed in populations of sensory neurons, where it mediates the sodium-activated potassium current (IKNa) and modulates neuronal activity. Previous studies suggest that Slack is involved in the processing of neuropathic pain. However, mechanisms underlying the regulation of Slack activity in this context are poorly understood. Using whole-cell patch-clamp recordings we found that Slack-mediated IKNa in sensory neurons of mice is reduced after peripheral nerve injury, thereby contributing to neuropathic pain hypersensitivity. Interestingly, Slack is closely associated with ATP-sensitive P2X3 receptors in a population of sensory neurons. In vitro experiments revealed that Slack-mediated IKNa may be bidirectionally modulated in response to P2X3 activation. Moreover, mice lacking Slack show altered nocifensive responses to P2X3 stimulation. Our study identifies P2X3/Slack signaling as a mechanism contributing to hypersensitivity after peripheral nerve injury and proposes a potential novel strategy for treatment of neuropathic pain.

Sodium-activated potassium channels moderate excitability in vascular smooth muscle.

We report that a sodium-activated potassium current, IKNa , has been inadvertently overlooked in both conduit and resistance arterial smooth muscle cells. IKNa is a major K+ resting conductance and is absent in cells of IKNa knockout (KO) mice. The phenotype of the IKNa KO is mild hypertension, although KO mice react more strongly than wild-type with raised blood pressure when challenged with vasoconstrictive agents. IKNa is negatively regulated by angiotensin II acting through Gαq protein-coupled receptors. In current clamp, KO arterial smooth muscle cells have easily evoked Ca2+ -dependent action potentials. Although several potassium currents have been reported to play a role in arterial smooth muscle (ASM), we find that one of the largest contributors to membrane conductance in both conduit and resistance ASMs has been inadvertently overlooked. In the present study, we show that IKNa , a sodium-activated potassium current, contributes a major portion of macroscopic outward current in a critical physiological voltage range that determines intrinsic cell excitability; IKNa is the largest contributor to ASM cell resting conductance. A genetic knockout (KO) mouse strain lacking KNa channels (KCNT1 and KCNT2) shows only a modest hypertensive phenotype. However, acute administration of vasoconstrictive agents such as angiotensin II (Ang II) and phenylephrine results in an abnormally large increase in blood pressure in the KO animals. In wild-type animals Ang II acting through Gαq protein-coupled receptors down-regulates IKNa , which increases the excitability of the ASMs. The complete genetic removal of IKNa in KO mice makes the mutant animal more vulnerable to vasoconstrictive agents, thus producing a paroxysmal-hypertensive phenotype. This may result from the lowering of cell resting K+ conductance allowing the cells to depolarize more readily to a variety of excitable stimuli. Thus, the sodium-activated potassium current may serve to moderate blood pressure in instances of heightened stress. IKNa may represent a new therapeutic target for hypertension and stroke.

GABA-B Controls Persistent Na+ Current and Coupled Na+-Activated K+ Current.

The GABA-B receptor is densely expressed throughout the brain and has been implicated in many CNS functions and disorders, including addiction, epilepsy, spasticity, schizophrenia, anxiety, cognitive deficits, and depression, as well as various aspects of nervous system development. How one GABA-B receptor is involved in so many aspects of CNS function remains unanswered. Activation of GABA-B receptors is normally thought to produce inhibitory responses in the nervous system, but puzzling contradictory responses exist. Here we report that in rat mitral cells of the olfactory bulb, GABA-B receptor activation inhibits both the persistent sodium current (INaP) and the sodium-activated potassium current (IKNa), which is coupled to it. We find that the primary effect of GABA-B activation is to inhibit INaP, which has the secondary effect of inhibiting IKNa because of its dependence on persistent sodium entry for activation. This can have either a net excitatory or inhibitory effect depending on the balance of INaP/IKNa currents in neurons. In the olfactory bulb, the cell bodies of mitral cells are densely packed with sodium-activated potassium channels. These channels produce a large IKNa which, if constitutively active, would shunt any synaptic potentials traversing the soma before reaching the spike initiation zone. However, GABA-B receptor activation might have the net effect of reducing the IKNa blocking effect, thus enhancing the effectiveness of synaptic potentials.

Identity, expression and functional role of the sodium-activated potassium current in vestibular ganglion afferent neurons

Vestibular afferent neurons (VANs) transmit information from the vestibular end organs to the central nuclei. This information is encoded within the firing pattern of these cells and is heavily influenced by the K+ conductances expressed by vestibular neurons. In the present study, we describe the presence of a previously unidentified Na+-activated K+ conductance (KNa) in these cells. We observed that the blocking of Na+ channels by tetrodotoxin (TTX) or the substitution of choline for Na+ in the extracellular solution during voltage clamp pulses resulted in the reduction of a sustained outward current that was dependent on the Na+ current. Furthermore, increases in the intracellular concentration of Na+ that were made by blocking the Na+/K+ ATPase with ouabain increased the amplitude of the outward current, and reduction of the intracellular Cl− concentration reduced the TTX-sensitive outward current. The substitution of Li+ for Na+ in the extracellular solution significantly reduced the amplitude of the outward current in voltage clamp pulses and decreased the afterhyperpolarization (AHP) of the action potentials in current clamp experiments. These electrophysiological results are consistent with the presence of mRNA transcripts for the KNa subunits Slick and Slack in the vestibular ganglia and in the sensory epithelium, which were detected using reverse-transcription polymerase chain reaction (RT-PCR). These results are also consistent with the immunolabeling of Slick and Slack protein in isolated vestibular neurons, in the vestibular ganglion and in the vestibular sensory epithelium. These results indicate that KNa channels are expressed in VANs and in their terminals. Furthermore, these data indicate that these channels may contribute to the firing pattern of vestibular neurons.

Adenosine modulation of potassium currents in postganglionic neurones of cultured avian ciliary ganglia

1. Potassium currents in cultured postganglionic neurones of avian ciliary ganglia were analysed under whole-cell voltage clamp and their modulation by adenosine determined. 2. In the presence of tetrodotoxin (200 nM), and with moderate holding potentials (Vh = -40 mV), the steady-state current-voltage (I/V) curve was N-shaped over the range from -70 mV to +155 mV. CsCl (1 M) blocked the current, indicating that it was carried by K+. If Ca2+ influx was blocked by CdCl2 (500 microM) then the outward current was reduced and the N-shaped I-V curve lost, indicating the presence of a calcium-activated potassium current (IK(Ca)); the remaining current, due to the delayed rectifier (IK), increased with depolarization up to about a conductance of 10 nS near + 50 mV. This IK was 50% activated at about +20 mV and 50% inactivated at about -50 mV. Adenosine (10 microM) had similar affects on the N-shaped I/V curve as did CdCl2, indicating that it blocked IK(Ca). However, adenosine had little affect on the steady-state current in the presence of CdCl, indicating that it did not much affect IK. 3. In the presence of tetrodotoxin (200 nM), a large inward current occurred for large hyperpolarizations from a Vh = -50 mV. This inward rectifying current (IIR) had a reversal potential near EK and showed 50% activation at about -130 mV. Adenosine (10 microM) reduced IIR, by as much as 50% at large hyperpolarizations beyond -80 mV. 4. Relaxations of the outward current on hyperpolarization from Vh = -30mV were blocked by carbachol (10 microM), had a reversal potential near EK, and an I/V curve typical of 1M currents. These currents were little affected by adenosine (10 microM). 5. A fast transient outward current, due to depolarizing pulses from a large Vh = -110mV was observed in the presence of tetrodotoxin (200 nM). This had the characteristics of an IA current as it could be blocked with 4-aminopyridine (5 mM) and was 50% activated at about -20 mV and 50% inactivated at about -94 mV. The IA current was reduced by 42% at a depolarization of -20 mV by adenosine (10 microM). 6. Many neurones possessed a fast transient outward current that was blocked by tetrodotoxin (200nM). This current could be blocked with 4-aminopyridine (5mM); it therefore has the characteristics of a sodium-activated potassium current ('K(Na)). This IK(Na) was unaffected by adenosine (1O microM). 7. These results are discussed in relation to the role of adenosine in blocking Ca2 + channels and thereby modifying calcium-dependent components of K+ currents.

Sodium-activated potassium current in mouse diaphragm

The mouse diaphragm muscle fiber was studied using the loose patch clamp technique. The voltage gated sodium currents were evoked by step pulses from a holding potential of about −70 mV. Following the activation of the sodium current, a very large and fast outward current was evoked. The sensitivity of this current to 4-aminopyridine and tetraethylammonium indicates the potassium ion as the possible carrier for the channel. Furthermore, the sensitivity to tetrodotoxin and extracellular sodium demonstrated the sodium dependence of this current.

Potassium current activated by intracellular sodium in quail trigeminal ganglion neurons.

Whole-cell voltage clamp and single-channel recordings were performed on cultured trigeminal ganglion neurons from quail embryos in order to study a sodium-activated potassium current (KNa). When KNa was activated by a step depolarization in voltage clamp, there was a proportionality between KNa and INa at all voltages between the threshold of INa and ENa. Single-channel recordings indicated that KNa could be activated already by 12 mM intracellular sodium and was almost fully activated at 50 mM sodium. 100 mM lithium, 100 mM choline, or 5 microM calcium did not activate KNa. The relationship between the probability for the channel to be open (Po) vs. the sodium concentration and the relationship of KNa open time-distributions vs. the sodium concentration suggest that two to three sodium ions bind cooperatively before KNa channels open. KNa channels were sensitive to depolarization; at 12 mM sodium, a 42-mV depolarization caused an e-fold increase in Po. Under physiological conditions, the conductance of the KNa channel was 50 pS. This conductance increased to 174 pS when the intra- and extracellular potassium concentrations were 75 and 150 mM, respectively.

Sodium-activated potassium current in cultured avian neurones

An important characteristic of excitable cells is their ability to repolarize rapidly after a depolarizing event such as a sodium action potential. This ability is caused by several fast potassium currents such as IA and IK (ref. 1) and possibly also a fast calcium-dependent potassium current (IK(Ca))2. Suppression of only one of the potassium channels by mutation or drugs can lead to anomalous behaviour in the animal2 or to epileptic disharges3,4 in in vitro preparations. Fast potassium currents vary in their inactivation properties1, and this may be the reason why many types of potassium currents are required for the adequate functioning of neurones in various physiological conditions. Here we describe a neuronal potassium current that is activated by intracellular sodium. This current can probably be triggered by the sodium entering the neurone during a single action potential and shows little voltage inactivation. It could therefore participate in the fast repolarization process in situations where other potassium currents are partially inactivated. Because this current will be activated by any process leading to sufficient entry of sodium, it may also be triggered by sodium-mediated excitatory synaptic events. This could help to control the level of excitability in response to synaptic inputs. A sodium-dependent potassium current was recently described in heart cells5, but it was suggested that this may only become activated in pathological conditions.