Transient Potassium Current Research Articles

Interleukin-10 inhibits angiotensin II-induced decrease in neuronal potassium current

Previously we demonstrated that viral-mediated increased expression of the anti-inflammatory cytokine interleukin-10 within the paraventricular nucleus of the hypothalamus significantly reduces blood pressure in normal rats made hypertensive by infusion of angiotensin II. However, the exact cellular locus of this interleukin-10 action within the paraventricular nucleus is unknown. In the present study we tested whether interleukin-10 exerts direct effects at its receptors located on hypothalamic neurons to offset the neuronal excitatory actions of angiotensin II via its type 1 receptors. The results indicated the presence of immunoreactive interleukin-10 receptors on neurons in normal rat paraventricular nucleus and that receptors for this cytokine were also expressed in neurons cultured from the hypothalamus. Patch-clamp electrophysiological recordings from these cultures revealed that extracellular application of interleukin-10 alone did not exert any alterations in neuronal membrane delayed rectifier or transient potassium currents. However, angiotensin II elicited a significant decrease in delayed rectifier potassium current, an effect that was abolished by interleukin-10 application. Since decreases in delayed rectifier potassium current contribute to increased neuronal excitability, this result is consistent with a direct inhibitory action of interleukin-10 on angiotensin-induced excitation of hypothalamic neurons. As such, these data are the first indication of a neuronal locus of action of interleukin-10 to temper the actions of angiotensin II in the hypothalamus.

Isoflurane depolarizes bronchopulmonary C neurons by inhibiting transient A-type and delayed rectifier potassium channels

Inhalation of isoflurane (ISO), a widely used volatile anesthetic, can produce clinical tachypnea. In dogs, this response is reportedly mediated by bronchopulmonary C-fibers (PCFs), but the relevant mechanisms remain unclear. Activation of transient A-type potassium current (IA) channels and delayed rectifier potassium current (IK) channels hyperpolarizes neurons, and inhibition of both channels by ISO increases neural firing. Due to the presence of these channels in the cell bodies of rat PCFs, we determined whether ISO could stimulate PCFs to produce tachypnea in anesthetized rats, and, if so, whether this response resulted from ISO-induced depolarization of the pulmonary C neurons via the inhibition of IA and IK. We recorded ventilatory responses to 5% ISO exposure in anesthetized rats before and after blocking PCF conduction and the responses of pulmonary C neurons (extracellularly recorded) to ISO exposure. ISO-induced (1mM) changes in pulmonary C neuron membrane potential and IA/IK were tested using the perforated patch clamp technique. We found that: (1) ISO inhalation evoked a brief tachypnea (∼7s) and that this response disappeared after blocking PCF conduction; (2) the ISO significantly elevated (by 138%) the firing rate of most pulmonary C neurons (17 out of 21) in the nodose ganglion; and (3) ISO perfusion depolarized the pulmonary C neurons in the vitro and inhibited both IA and IK, and this evoked-depolarization was largely diminished after blocking both IA and IK. Our results suggest that ISO is able to stimulate PCFs to elicit tachypnea in rats, at least partly, via inhibiting IA and IK, thereby depolarizing the pulmonary C neurons.

Activation of high and low affinity dopamine receptors generates a closed loop that maintains a conductance ratio and its activity correlate

Neuromodulators alter network output and have the potential to destabilize a circuit. The mechanisms maintaining stability in the face of neuromodulation are not well described. Using the pyloric network in the crustacean stomatogastric nervous system, we show that dopamine (DA) does not simply alter circuit output, but activates a closed loop in which DA-induced alterations in circuit output consequently drive a change in an ionic conductance to preserve a conductance ratio and its activity correlate. DA acted at low affinity type 1 receptors (D1Rs) to induce an immediate modulatory decrease in the transient potassium current (IA) of a pyloric neuron. This, in turn, advanced the activity phase of that component neuron, which disrupted its network function and thereby destabilized the circuit. DA simultaneously acted at high affinity D1Rs on the same neuron to confer activity-dependence upon the hyperpolarization activated current (Ih) such that the DA-induced changes in activity subsequently reduced Ih. This DA-enabled, activity-dependent, intrinsic plasticity exactly compensated for the modulatory decrease in IA to restore the IA:Ih ratio and neuronal activity phase, thereby closing an open loop created by the modulator. Activation of closed loops to preserve conductance ratios may represent a fundamental operating principle neuromodulatory systems use to ensure stability in their target networks.

Tonic Dopamine Induces Persistent Changes in the Transient Potassium Current through Translational Regulation

Neuromodulatory effects can vary with their mode of transmission. Phasic release produces local and transient increases in dopamine (DA) up to micromolar concentrations. Additionally, since DA is released from open synapses and reuptake mechanisms are not nearby, tonic nanomolar DA exists in the extracellular space. Do phasic and tonic transmissions similarly regulate voltage-dependent ionic conductances in a given neuron? It was previously shown that DA could immediately alter the transient potassium current (I(A)) of identified neurons in the stomatogastric ganglion of the spiny lobster Panulirus interruptus. Here we show that DA can also persistently alter I(A), and that the immediate and persistent effects of DA oppose one another. The lateral pyloric (LP) neuron exclusively expresses type 1 DA receptors (D1Rs). Micromolar DA produces immediate depolarizing shifts in the voltage dependence of LP I(A), whereas tonic nanomolar DA produces a persistent increase in LP I(A) maximal conductance (G(max)) through a translation-dependent mechanism involving target of rapamycin (TOR). The pyloric dilator (PD) neuron exclusively expresses D2Rs. Micromolar DA produces an immediate hyperpolarizing shift in PD I(A) voltage dependence of activation, whereas tonic DA persistently decreases PD I(A) G(max) through a translation-dependent mechanism not involving TOR. The persistent effects on I(A) G(max) do not depend on LP or PD activity. These data suggest a role for tonic modulators in the regulation of voltage-gated ion channel number; and furthermore, that dopaminergic systems may be organized to limit the amount of change they can impose on a circuit.

Effects of Resibufogenin and Cinobufagin on voltage-gated potassium channels in primary cultures of rat hippocampal neurons

Outward delayed rectifier potassium channel and outward transient potassium channel have multiple important roles in maintaining the excitability of hippocampal neurons. The present study investigated the effects of two bufadienolides, Resibufogenin (RBG) and Cinobufagin (CBG), on the outward delayed rectifier potassium current (IK) and outward transient potassium current (IA) in rat hippocampal neurons. RBG and CBG have similar structures and both were isolated from the venom gland of toad skin. RBG inhibited both IK and IA, whereas CBG inhibited IK without noticeable effect on IA. Moreover, at 1μM concentration both RBG and CBG could alter some channel kinetics and gating properties of IK, such as steady-state activation and inactivation curves, open probability and time constants. These findings suggested that IK is probably a target of bufadienolides, which may explain the mechanisms of bufadienolides’ pathological effects on central nervous system.

Differential contribution of A-type potassium currents in shaping neuronal responses to synaptic input

Neurons receive input from thousands of excitatory and inhibitory synapses, but they are not merely passive receivers and transmitters of information. The intrinsic properties of neurons, and more specifically the collection of ion channels expressed, influence a cell's excitability and integration processes. The role of different voltage-dependent transient (A-type) and persistent (delayed rectifier) potassium currents in shaping the activity of neurons in response to synaptic input is not well understood. We developed a biophysical model incorporating realistic synaptic input [1] to investigate the specific roles played by distinct A-type potassium channels encoded by different genes, namely Shaker/Kv1 and Shal/Kv4. We find that these two channel types, which are functionally distinguished by their inactivation properties, exert different control mechanisms on the transitions between rest and repetitive spiking in neurons. Bifurcation analysis is used to characterize the influence of each channel and the overall behavior of the membrane as a function of the relative presence of the two A-type channels. We found that the influence of each channel type depends on the resting potential of the neuron, suggesting that modulation and/or network states may shift their relative contribution. For example, at a particular membrane potential, Shal/Kv4 channels are more likely to mediate delays to first spike in response to excitatory input. Overall, this work increases our understanding of the interaction between intrinsic properties and synaptic input in creating neuronal response profiles. Further, we provide a link between relative gene expression and bifurcation structures in dynamical systems modeling neuronal membranes.

Restoring ion channel pathology by parameter optimization

In diseases of the brain, the distribution and properties of ion channels display deviations from healthy control subjects. We studied two cases of ion channel alterations related to epileptogenesis. The first case of ion channel alteration represents an enhanced sodium current, the second case addresses the run down of conductance of the transient potassium current, KA. In previous studies we have shown that KA reduce highly synchronized synaptic input while minimally affect semi-synchronized input (1). The KA channel may therefore function as a protective mechanism against synchronous input involved in seizures. In this study we investigate if modulatory substances which targets the KA can functionally corrected the two pathological models.

Control of firing patterns by two transient potassium currents: leading spike, latency, bistability.

Transient potassium currents distinctively affect firing properties, particularly in regulating the latency before repetitive firing. Pyramidal cells of the dorsal cochlear nucleus (DCN) have two transient potassium currents, I(Kif) and I(Kis), fast and slowly inactivating, respectively, and they exhibit firing patterns with dramatically variable latencies. They show immediate repetitive firing, or only after a long latency with or without a leading spike, the so-called pauser and buildup patterns. We consider a conductance-based, ten-variable, single-compartment model for the DCN pyramidal cells (Kanold and Manis 2001). We develop and analyze a reduced three-variable integrate-and-fire model (KM-LIF) which captures the qualitative firing features. We apply dynamical systems methods to explain the underlying biophysical and mathematical mechanisms for the firing behaviors, including the characteristic firing patterns, the latency phase, the onset of repetitive firing, and some discontinuities in the timing of latency duration (e.i. first spike latency and first inter spike interval). Moreover, we obtain new insights associated with the leading spike by phase plane analysis. We further demonstrate the effects of possible heterogeneity of I(Kis). The latency before repetitive firing can be controlled to cover a large range by tuning of the relative amounts of I(Kif) and I(Kis). Finally, we find for the full system robust bistability when enough I(Kis) is present.

Cell Specific Dopamine Modulation of the Transient Potassium Current in the Pyloric Network by the Canonical D1 Receptor Signal Transduction Cascade

Dopamine (DA) modifies the motor pattern generated by the pyloric network in the stomatogastric ganglion (STG) of the spiny lobster, Panulirus interruptus, by directly acting on each of the circuit neurons. The 14 pyloric neurons fall into six cell types, and DA actions are cell type specific. The transient potassium current mediated by shal channels (I(A)) is a common target of DA modulation in most cell types. DA shifts the voltage dependence of I(A) in opposing directions in pyloric dilator (PD) versus lateral pyloric (LP) neurons. The mechanism(s) underpinning cell-type specific DA modulation of I(A) is unknown. DA receptors (DARs) can be classified as type 1 (D1R) or type 2 (D2R). D1Rs and D2Rs are known to increase and decrease intracellular cAMP concentrations, respectively. We hypothesized that the opposing DA effects on PD and LP I(A) were due to differences in DAR expression patterns. In the present study, we found that LP expressed somatodendritic D1Rs that were concentrated near synapses but did not express D2Rs. Consistently, DA modulation of LP I(A) was mediated by a Gs-adenylyl cyclase-cAMP-protein kinase A pathway. Additionally, we defined antagonists for lobster D1Rs (flupenthixol) and D2Rs (metoclopramide) in a heterologous expression system and showed that DA modulation of LP I(A) was blocked by flupenthixol but not by metoclopramide. We previously showed that PD neurons express D2Rs, but not D1Rs, thus supporting the idea that cell specific effects of DA on I(A) are due to differences in receptor expression.

Actions of bis(7)-tacrine and tacrine on transient potassium current in rat DRG neurons and potassium current mediated by K V4.2 expressed in Xenopus oocyte

Bis(7)-tacrine [bis(7)-tetrahydroaminacrine] is a dimeric AChE inhibitor derived from tacrine with a potential to treat Alzheimer's disease. Actions of bis(7)-tacrine on ligand-gated ion channels and voltage-gated cation channels have been identified on neurons of both central and peripheral nervous systems. In the present study, the effect of bis(7)-tacrine was investigated on the K V4.2 encoded potassium currents expressed in Xenopus oocytes and the transient A-type potassium current ( I K(A)) on rat DRG neurons. Bis(7)-tacrine suppressed recombinant Kv4.2 potassium channels in a concentration-dependent manner, with IC 50 value of 0.53 ± 0.13 μM. Tacrine also inhibited Kv4.2 channels, but with a much lower potency (IC 50 74 ± 15 μM).The possible mechanisms underlying the inhibition on potassium currents by bis(7)-tacrine/tacrine could be that inactivation of the transient potassium currents was accelerated and recovery of the native or Kv4.2 expressed potassium currents was suppressed by bis(7)-tacrine/tacrine.

Angiotensin II increases excitability and inhibits a transient potassium current in vagal primary sensory neurons

The octapeptide angiotensin II (ANG II) plays a pivotal role in the maintenance of blood pressure by activating ANG II receptors located in variety of cell types including neurons housed in the central nervous system (CNS) and in the peripheral nervous system (PNS). ANG II (100 nM) blocked spike frequency accommodation (SFA) recorded with whole-cell patch technique in acutely isolated nodose ganglion neurons (NGN) from adult rats. ANG II increased the frequency of action potentials (AP) produced by supramaximal 500 ms depolarizing currents recorded in both tonic (16 Hz vs. 58 Hz, control vs. ANG II perfusion respectively, n=9) and phasic (1Hz vs. 38 Hz, n=13) NGNs. ANG II produced no significant changes in: the resting membrane potential (-51 mV vs. -50 mV, n=65), AP overshoot (46 mV vs. 41 mV, n=25), AP undershoot (-65 mV vs. -61 mV, n=25), AP duration (1 ms vs. 1.2 ms, n=25), and AP threshold (-40 mV vs. -43 mV, n=19). CV-11974 (600 nM), a specific AT1 receptor antagonist, prevented ANG II-evoked changes SFA (n=10). ANG II (100 nM) had no significant effect on total outward potassium current (I(K)) but inhibited a fast activating and fast inactivating I(K) recorded in the presence of TEA. A kinetically similar I(K) was also inhibited by 4-AP (3mM). In phasic NGNs, 4-AP occluded the effects of 100 nM ANG II on SFA. Our results indicate that ANG II can block an A-type of I(K) and that this effect may underlie the ANG II-mediated change in SFA.

Hypercapnia inhibits both transient and sustained potassium currents in chemosensitive neurons from neonatal rat locus coeruleus (LC)

Increased ventilation is stimulated by an increase in CO2/H+, which is detected by peripheral chemoreceptors and central chemosensitive neurons. We studied CO2/H+‐sensitive K+ channels in rat LC neurons. We hypothesize that multiple ion channels are involved in the neuronal response to changes in CO2/H+, with K+ channels being an important component mediating this response. In this study, we used whole cell voltage clamp to test the effect of 15% CO2 on both transient and sustained K+ currents of LC neurons from neonatal rat brain slices. Neurons were clamped at ‐80mV. Voltage‐dependent K+ currents were evoked by various depolarizing pulses (300 ms duration) in 10 mV increments from ‐100mV to 50mV in the presence of TTX (1 μm) and Cd2+ (200 μm) to block Na+ and Ca2+ channels, respectively. Transient K+ currents were inhibited by 4‐amino pyridine (4‐AP; 5mM) and sustained K+ currents were inhibited by tetraethylammonium (TEA; 20mM). Five minutes after perfusion with aCSF equilibrated with 15% CO2, the conductances of both transient and sustained K+ channels were decreased by 28 ± 9.5% (n=3) and 19 ± 9.6% (n=3), respectively. These results support the hypothesis that hypercapnia increases the firing frequency of LC neurons through depolarization due to inhibition of multiple K+ channels, including transient and sustained K+ conductances.Supported by NIH Grant R01‐HL56683‐11.

Ionic currents in hair cells dissociated from frog semicircular canals after preconditioning under microgravity conditions

The effects of microgravity on the biophysical properties of frog labyrinthine hair cells have been examined by analyzing calcium and potassium currents in isolated cells by the patch-clamp technique. The entire, anesthetized frog was exposed to vector-free gravity in a random positioning machine (RPM) and the functional modification induced on single hair cells, dissected from the crista ampullaris, were subsequently studied in vitro. The major targets of microgravity exposure were the calcium/potassium current system and the kinetic mechanism of the fast transient potassium current, I(A). The amplitude of I(Ca) was significantly reduced in microgravity-conditioned cells. The delayed current, I(KD) (a complex of I(KV) and I(KCa)), was drastically reduced, mostly in its I(KCa) component. Microgravity also affected I(KD) kinetics by shifting the steady-state inactivation curve toward negative potentials and increasing the sensitivity of inactivation removal to voltage. As concerns the I(A), the I-V and steady-state inactivation curves were indistinguishable under normogravity or microgravity conditions; conversely, I(A) decay systematically displayed a two-exponential time course and longer time constants in microgravity, thus potentially providing a larger K(+) charge; furthermore, I(A) inactivation removal at -70 mV was slowed down. Stimulation in the RPM machine under normogravity conditions resulted in minor effects on I(KD) and, occasionally, incomplete I(A) inactivation at -40 mV. Reduced calcium influx and increased K(+) repolarizing charge, to variable extents depending on the history of membrane potential, constitute a likely cause for the failure in the afferent mEPSP discharge at the cytoneural junction observed in the intact labyrinth after microgravity conditioning.

Excitability of small-diameter trigeminal ganglion neurons by 5-HT is mediated by enhancement of the tetrodotoxin-resistant sodium current due to the activation of 5-HT4 receptors and/or by the inhibition of the transient potassium current

The aims of the present study were to investigate whether the activation of the 5-HT receptor subtypes (5-HT4 and 5-HT3) acted significantly on the modification of the tetrodotoxin-resistant sodium current (INaR) in small-sized rat trigeminal ganglion (TG) neurons and whether the inhibition of the transient K+ current (IA) contributed to the excitability in those neurons. 5-HT applications in at concentrations ranging from 0.01–10 μM significantly increased the peak INaR. One micromolar 5-HT application caused the greatest increase in the peak INaR amplitude accompanied by a hyperpolarizing shift in the activation curve. A similar modification of INaR properties was also obtained via the application of the 5-HT4 receptor agonist, RS 67333, in concentrations ranging from 0.001–1 μM. The largest effects of 5-HT (1 μM) and RS 67333 (0.1 μM) on the modification of INaR were abolished by pretreatment with ICS 205–930 (a 5-HT3/4 receptor antagonist, 10 μM), which showed no significant effect on the baseline INaR. However, ICS 205–930 application at 30 μM caused a significant decrease in the baseline INaR. Phenylbiguanide (a 5-HT3 receptor agonist) did not significantly alter INaR properties when applied in concentrations ranging from 1 to 100 μM. The application of 0.1 μM RS 67333 decreased the transient K+ current (IA) by approximately 31%. The threshold for action potential generation was significantly lower after the application of 0.1 μM RS 67333. Furthermore, 0.1 μM RS 67333 application increased the number of action potentials and the resting membrane potential got more positive, but it decreased the duration of depolarization phase of action potential. In addition, neither the additional application of 1 μM 5-HT in the presence of 10 μM forskolin, a stimulator of adenylyl cyclase, nor the opposite applications of 5-HT and forskolin caused the enhancement of increased INaR, which indicates the presence of an ‘occluding effect.’ These results suggest that the 5-HT-induced modification of INaR is mediated by the activation of 5-HT4 receptors, involving a cAMP-dependent signaling pathway, and that the inhibition of IA following the application of a 5-HT4 receptor agonist also contributes to the increased number of action potentials.

FMRFamide MODULATES OUTWARD POTASSIUM CURRENTS IN MOUSE OLFACTORY SENSORY NEURONS

1. The olfactory system can detect the presence of low concentrations of odourant molecules and discriminate even slight differences among molecules with a very similar chemical structure. The detection and discrimination of odourants are correlated with the electrophysiology of the olfactory sensory neurons. To get a better understanding of the molecular mechanisms of olfactory transduction, it is therefore of considerable importance to obtain electrophysiological recordings of olfactory sensory neurons. FMRFamide (Phe-Met-Arg-Phe-NH(2)), secreted from the nerve terminals of the nasal cavity, has been suggested to act as a neurotransmitter or neuromodulator, playing an important role in modulating the activity of olfactory receptor neurons. Its effects on voltage-dependent potassium currents in the mouse olfactory sensory neurons were investigated in the present study using the whole-cell patch-clamp technique. 2. Olfactory sensory neurons were isolated from the Kunming Mouse (KM) mouse olfactory epithelium. Different protocols were applied to obtain delayed-rectifier potassium current (I(K)) and fast transient potassium current (I(A)). The effects of FMRFamide on the outward potassium currents, including I(K) and I(A), in mouse olfactory sensory neurons were investigated. 3. We found that FMRFamide (5 micromol/L) increased the magnitude of I(K). However no effect on I(A) was observed. The activation dynamics of both currents were not changed by FMRFamide. 4. In conclusion, FMRFamide may play a role in the modulation of peripheral olfactory signals by regulating I(K). This modulation may shorten the phase of the fast repolarization of the action potential in mouse olfactory sensory neurons and increase the excitability of the neuronal membrane.

Peroxynitrite donor impairs excitability of hippocampal CA1 neurons by inhibiting voltage-gated potassium currents

Nitric oxide (NO) reacts rapidly with superoxide anion and produces peroxynitrite (ONOO −), which is a highly reactive free radical that has been shown to mediate much of the toxicity of NO. ONOO − has also been implicated as playing a role in the pathology of various brain disorders. The aim of this study was to investigate the actions of ONOO − on voltage-activated potassium currents and its effect on membrane excitability in hippocampal CA1 neurons. SIN-1(3-morpholino-sydnonimine), which leads to the simultaneous generation of superoxide anion and NO, and then forms the highly reactive species ONOO −, induced a dose-dependent inhibition in amplitudes of transient potassium currents ( I A) and delayed rectifier potassium currents ( I K). SIN-1 (500 μM) produced a hyperpolarizing shift in the activation–voltage curve of both I A and I K and delayed the recovery of I A from inactivity, but did not have a marked effect on the inactivation parameters of I A. However, co-treatment with SIN-1 and Mn(III)tetrakis(4-benzoic acid)porphyrin chloride (MnTBAP, 100 μM), an ONOO − scavenger, had no effects on I A and I K currents. SIN-1 (500 μM) led to increase in action potential duration and decrease in action potential firing rate. The results suggest that changes of I A and I K currents induced by ONOO − in hippocampal neurons have influence on excitability of hippocampal CA1 neurons, which are related to its neurotoxicity.

Roles of Ionic Currents in Lamprey CPG Neurons: A Modeling Study

The spinal network underlying locomotion in the lamprey consists of a core network of glutamatergic and glycinergic interneurons, previously studied experimentally and through mathematical modeling. We present a new and more detailed computational model of lamprey locomotor network neurons, based primarily on detailed electrophysiological measurements and incorporating new experimental findings. The model uses a Hodgkin-Huxley-like formalism and consists of 86 membrane compartments containing 12 types of ion currents. One of the goals was to introduce a fast, transient potassium current (K(t)) and two sodium-dependent potassium currents, one faster (K(NaF)) and one slower (K(NaS)), in the model. Not only has the model lent support to the interpretation of experimental results but it has also provided predictions for further experimental analysis of single-network neurons. For example, K(t) was shown to be one critical factor for controlling action potential duration. In addition, the model has proved helpful in investigating the possible influence of the slow afterhyperpolarization on repetitive firing during ongoing activation. In particular, the balance between the simulated slow sodium-dependent and calcium-dependent potassium currents has been explored, as well as the possible involvement of dendritic conductances.

Vitamins C and E Modulate Neuronal Potassium Currents

We investigated the effects of vitamins C and E on the delayed-rectifier potassium current (IK(DR)), which is important in repolarizing the membrane potential, and on the transient A-type potassium current (IK(A)), which regulates neuronal firing frequency. The whole-cell patch-clamp technique was used to measure the currents from cultured Drosophila neurons derived from embryonic neuroblasts. The membrane potential was stepped to different voltages between -40 and +60 mV from a holding potential of -80 mV. IK(DR) and IK(A) measured in the vitamin C-containing solution (IK(DR) 305 +/- 16 pA, IK(A) 11 +/- 2 pA) were smaller than those measured in the control solution (488 +/- 21 pA, IK(A )28 +/- 3 pA). By contrast, IK(DR) and IK(A) measured in the vitamin E-containing solution (IK(DR) 561 +/- 21 pA, IK(A )31 +/- 3 pA) were greater than those measured in the control solution (422 +/- 15 pA, 17 +/- 2 pA). These results indicate that vitamins C and E can modulate potassium current amplitudes and possibly lead to altered neuronal excitability.

Protein kinase C activation-induced increases of neural activity are enhanced in the hypothalamus of spontaneously hypertensive rats

We have previously reported that some neurons in the anterior hypothalamic area (AHA) are tonically activated by endogenous angiotensins in rats and that activities of these angiotensin II-sensitive neurons in the AHA are enhanced in spontaneously hypertensive rats (SHR). In addition, neural activations induced by both angiotensin II and glutamate were enhanced in the AHA of SHR. In this study, we examined whether intracellular neural activation mechanisms via protein kinase C (PKC) and a potassium channel are altered in angiotensin II-sensitive neurons in the AHA of SHR. Male 15- to 16-week-old SHR and age-matched Wistar–Kyoto rats (WKY) and Wistar rats were anesthetized and artificially ventilated. Extracellular potentials were recorded from single neurons in the AHA. Pressure application of the PKC activator phorbol 12-myristate 13-acetate (PMA) onto angiotensin II-sensitive neurons in the AHA of Wistar rats increased their firing rate. The increase of unit activity by PMA was inhibited by the potent inhibitor of PKC, 1-(5-isoquinolinesulfonyl)-2-methylpiperazine dihydrochloride (H-7), but not by the weak PKC inhibitor, N-(2-guanidinoethyl)-5-isoquinolinesulfonamide hydrochloride (HA1004). The increase of unit firing by PMA was enhanced in SHR as compared with WKY. Pressure application of H-7 alone decreased the basal firing activity of angiotensin II-sensitive neurons in SHR but not in WKY. HA1004 did not affect the basal firing activity of angiotensin II-sensitive neurons in SHR. Angiotensin II-induced increases of firing rate in AHA neurons were inhibited by H-7 and the inhibition by H-7 was enhanced in SHR as compared with WKY. Pressure application of 4-aminopyridine, a blocker of the transient potassium current, onto angiotensin II-sensitive neurons increased their firing rate and the increase of unit firing rate was almost the same in WKY and SHR. These findings indicate that activation of PKC increases neural activity in angiotensin II-sensitive neurons in the AHA and that this PKC activation-induced increase of neural activity is enhanced in the AHA of SHR. It seems likely that the enhanced PKC activation effect is responsible for the enhanced basal neural activity seen in the AHA of SHR.

Biophysical properties of the silent and activated rat sympathetic neuron following denervation

A biophysical description of the denervated rat sympathetic neuron is reported, obtained by the two-electrode voltage-clamp technique in mature intact superior cervical ganglia in vitro. At membrane potential values negative to −50 mV, the normal, quiescent neuron displays voltage-dependent K and Cl conductances; following direct or synaptic stimulation (15Hz for 10 s), the neuron moves to a new resting state characterized by increased amplitude and voltage dependence of Cl conductance. Denervation produces two main effects: 1) resting Cl conductance gradually increases while its voltage-dependence decreases; by 30 days a high-conductance resting state prevails, almost independent of membrane potential in the −50/−110 mV range; 2) the increase in amplitude and voltage-dependence of Cl conductance, produced by direct stimulation in control neurons, is less marked in denervated neurons, and is observed over an increasingly small range of membrane potentials. Thirty days after denervation, the prevailing high-conductance resting state appears virtually insensitive to changes in membrane potential and stimulation. Voltage-dependent potassium currents involved in spike electrogenesis (the delayed compound potassium current and the fast transient potassium current) exhibit an early drastic decrease in peak amplitude in the denervated neuron; the effect is largely reversed after 6 days. Remarkable changes in fast transient potassium current kinetics occur following denervation: the steady-state inactivation curve shifts by up to +15 mV toward positive potential and voltage sensitivity of inactivation removal becomes more steep. A comprehensive mathematical model of the denervated neuron is presented that fits the neuron behavior under current-clamp conditions. It confirms that neuronal excitability is tuned by the conductances (mostly chloride conductance) that control the resting membrane potential level, and by fast transient potassium current. Impairment of the latter reduces both inward threshold charge for firing and spike repolarization rate, and fast transient potassium current failure cancels the voltage dependence of both processes.