Rectifying Potassium Conductance Research Articles

Opioid inhibition of projection‐specific, respiratory‐related pontomedullary neurons

Opioids depress breathing by inhibition of inter‐connected respiratory nuclei in the pons and medulla. We have previously shown that mu opioid receptor (MOR) agonists hyperpolarize a population of neurons in the dorsolateral pons due to activation of G protein‐mediated inwardly rectifying potassium conductance. However, we do not know the projection target of MOR‐expressing, dorsolateral pontine neurons, if they project at all. Here, we show that MOR‐expressing, dorsolateral pontine neurons project to core respiratory nuclei in the ventrolateral medulla, identifying projection‐related heterogeneity of dorsolateral pontine neurons vulnerable to opioid inhibition. First, retrograde DIO‐eGFP injections into the ventrolateral medulla of oprm1‐cre mice showed that dorsolateral pontine neurons project to the Bötzinger complex (BötC), pre‐Bötzinger complex (preBötC), and rostral ventral respiratory group (rVRG), and are distinct from calcitonin gene‐related peptide (CGRP)‐expressing parabrachial neurons. Second, we performed whole‐cell, voltage‐clamp recordings from dorsolateral pontine Kölliker‐Fuse neurons that were retrogradely labeled by fluorescent bead injections into the ventrolateral medulla of wild‐type mice. MOR‐mediated outward currents were identified in dorsolateral pontine neurons that project to the BötC, preBötC, and rVRG, confirming that these projection neurons are functionally inhibited by MORs and vulnerable to opioid inhibition. Together, these approaches demonstrate that MOR‐expressing, dorsolateral pontine neurons project to medullary respiratory nuclei and further unravel mechanisms of opioid‐induced respiratory depression.

A population of in silico models identifies the interplay between Nav 1.8 conductance and potassium currents as key in regulating human dorsal root ganglion neuron excitability.

Background: The Nav 1.8 sodium channel has a key role in generating repetitive action potentials in nociceptive human dorsal root ganglion neurons. Nav 1.8 is differentiated from other voltage-gated sodium channels by its unusually slow inactivation kinetics and depolarised voltage-dependence of activation. These features are particularly pronounced in the human Nav 1.8 channel and allow the channel to remain active during repolarisation. Gain-of-function mutations in Nav 1.8 have been linked to neuropathic pain and selective blockers of Nav 1.8 have been developed as potential new analgesics. However, it is not well understood how modulating the Nav 1.8 conductance alters neuronal excitability and how this depends on the balance of other ion channels expressed by nociceptive neurons. Methods: To investigate this, we developed a novel computational model of the human dorsal root ganglion neuron and used it to construct a population of models that mimicked inter-neuronal heterogeneity in ionic conductances and action potential morphology Results: By simulating changes to the Nav 1.8 conductance in the population of models, we found that moderately increasing the Nav 1.8 conductance led to increased firing rate, as expected, but increasing Nav 1.8 conductance beyond an inflection point caused firing rate to decrease. We found that the delayed rectifier and M-type potassium conductances were also critical for determining neuronal excitability. In particular, altering the delayed rectifier potassium conductance shifted the position of the Nav 1.8 inflection point and therefore the relationship between Nav 1.8 conductance and firing rate. Conclusions: Our results suggest that the effects of modulating Nav 1.8 in a nociceptive neuron can depend significantly on other conductances, particularly potassium conductances.

Investigating the Cocaine-induced Reduction of Potassium Current on the Generation of Action Potentials Using a Computational Model.

Drugs of abuse, such as cocaine, affect different brain regions and lead to pathological memories. These abnormal memories may occur due to changes in synaptic transmissions or variations in synaptic properties of neurons. It has been shown that cocaine inhibits delayed rectifying potassium currents in affected brain regions and can create pathological memories.This study investigates how the change in the conductance of delayed rectifying potassium channels can affect the produced action potentials using a computational model. We present a computational model with different channels and receptors, including sodium, potassium, calcium, NMDARs, and AMPARs, which can produce burst-type action potentials. In the simulations, by changing the delayed rectifying potassium conductance bifurcation diagram is calculated. By decreasing the potassium current for a fixed stimulatory signal, burst-type action potentials can be generated. In the following and with a further reduction of potassium conductance, produced action potentials exhibit non-linear and even chaotic behaviors. Results show that for a specific range of potassium conductance, a chaotic regime emerges in produced action potentials. These chaotic oscillations may play a role in inducing abnormal memories. Cocaine consumption reduces the potassium current in affected cells.Decreasing the potassium currents elicits burst action potentials.Produced bursts might have chaotic behaviors.Chaotic oscillations might be related to the toxic effects of cocaine. Drugs of abuse such as cocaine can manipulate brain circuits and may form some pathological memories. These memories can lead to long-term addiction. Furthermore, these drugs also can have toxic effects on the cells. Researchers are looking for the mechanisms that can lead to abnormal memories and toxic effects of drugs. It seems that an efficient mechanism that can be used by drugs of abuse is the manipulation of potassium currents in the affected cells. Here, in a computational model, we have shown that changes in the conductance of delayed rectifying potassium channels can lead to nonlinear and even chaotic behaviors in the produced action potentials. These behaviors might have a role in drug toxic effects.

Two opposite voltage-dependent currents control the unusual early development pattern of embryonic Renshaw cell electrical activity.

Renshaw cells (V1R) are excitable as soon as they reach their final location next to the spinal motoneurons and are functionally heterogeneous. Using multiple experimental approaches, in combination with biophysical modeling and dynamical systems theory, we analyzed, for the first time, the mechanisms underlying the electrophysiological properties of V1R during early embryonic development of the mouse spinal cord locomotor networks (E11.5-E16.5). We found that these interneurons are subdivided into several functional clusters from E11.5 and then display an unexpected transitory involution process during which they lose their ability to sustain tonic firing. We demonstrated that the essential factor controlling the diversity of the discharge pattern of embryonic V1R is the ratio of a persistent sodium conductance to a delayed rectifier potassium conductance. Taken together, our results reveal how a simple mechanism, based on the synergy of two voltage-dependent conductances that are ubiquitous in neurons, can produce functional diversity in embryonic V1R and control their early developmental trajectory.

Alterations in Intrinsic and Synaptic Properties of Hippocampal CA1 VIP Interneurons During Aging.

Learning and memory deficits are hallmarks of the aging brain, with cortical neuronal circuits representing the main target in cognitive deterioration. While GABAergic inhibitory and disinhibitory circuits are critical in supporting cognitive processes, their roles in age-related cognitive decline remain largely unknown. Here, we examined the morphological and physiological properties of the hippocampal CA1 vasoactive intestinal peptide/calretinin-expressing (VIP+/CR+) type 3 interneuron-specific (I-S3) cells across mouse lifespan. Our data showed that while the number and morphological features of I-S3 cells remained unchanged, their firing and synaptic properties were significantly altered in old animals. In particular, the action potential duration and the level of steady-state depolarization were significantly increased in old animals in parallel with a significant decrease in the maximal firing frequency. Reducing the fast-delayed rectifier potassium or transient sodium conductances in I-S3 cell computational models could reproduce the age-related changes in I-S3 cell firing properties. However, experimental data revealed no difference in the activation properties of the Kv3.1 and A-type potassium currents, indicating that transient sodium together with other ion conductances may be responsible for the observed phenomena. Furthermore, I-S3 cells in aged mice received a stronger inhibitory drive due to concomitant increase in the amplitude and frequency of spontaneous inhibitory currents. These age-associated changes in the I-S3 cell properties occurred in parallel with an increased inhibition of their target interneurons and were associated with spatial memory deficits and increased anxiety. Taken together, these data indicate that VIP+/CR+ interneurons responsible for local circuit disinhibition survive during aging but exhibit significantly altered physiological properties, which may result in the increased inhibition of hippocampal interneurons and distorted mnemonic functions.

Dopamine D2L Receptor Deficiency Causes Stress Vulnerability through 5-HT1A Receptor Dysfunction in Serotonergic Neurons.

Mental disorders are caused by genetic and environmental factors. We here show that deficiency of an isoform of dopamine D2 receptor (D2R), D2LR, causes stress vulnerability in mouse. This occurs through dysfunction of serotonin [5-hydroxytryptamine (5-HT)] 1A receptor (5-HT1AR) on serotonergic neurons in the mouse brain. Exposure to forced swim stress significantly increased anxiety- and depressive-like behaviors in D2LR knock-out (KO) male mice compared with wild-type mice. Treatment with 8-OH-DPAT, a 5-HT1AR agonist, failed to alleviate the stress-induced behaviors in D2LR-KO mice. In forced swim-stressed D2LR-KO mice, 5-HT efflux in the medial prefrontal cortex was elevated and the expression of genes related to 5-HT levels was upregulated by the transcription factor PET1 in the dorsal raphe nucleus. Notably, D2LR formed a heteromer with 5-HT1AR in serotonergic neurons, thereby suppressing 5-HT1AR-activated G-protein-activated inwardly rectifying potassium conductance in D2LR-KO serotonergic neurons. Finally, D2LR overexpression in serotonergic neurons in the dorsal raphe nucleus alleviated stress vulnerability observed in D2LR-KO mice. Together, we conclude that disruption of the negative feedback regulation by the D2LR/5-HT1A heteromer causes stress vulnerability.SIGNIFICANCE STATEMENT Etiologies of mental disorders are multifactorial, e.g., interactions between genetic and environmental factors. In this study, using a mouse model, we showed that genetic depletion of an isoform of dopamine D2 receptor, D2LR, causes stress vulnerability associated with dysfunction of serotonin 1A receptor, 5-HT1AR in serotonergic neurons. The D2LR/5-HT1AR inhibitory G-protein-coupled heteromer may function as a negative feedback regulator to suppress psychosocial stress.

Thermal constraints on in vivo optogenetic manipulations.

A key assumption of optogenetics is that light only affects opsin-expressing neurons. However, illumination invariably heats tissue, and many physiological processes are temperature-sensitive. Commonly-used illumination protocols increased temperature by 0.2–2°C and suppressed spiking in multiple brain regions. In striatum, light delivery activated an inwardly-rectifying potassium conductance and biased rotational behavior. Thus, careful consideration of light delivery parameters is required, as even modest intracranial heating can confound interpretation of optogenetic experiments.

Experimental Cortical Spreading Depression Induces NMDA Receptor Dependent Potassium Currents in Microglia.

Cortical spreading depression (CSD) is a propagating event of neuronal depolarization, which is considered as the cellular correlate of the migraine aura. It is characterized by a change in the intrinsic optical signal and by a negative DC potential shift. Microglia are the resident macrophages of the CNS and act as sensors for pathological changes. In the present study, we analyzed whether microglial cells might sense CSD by recording membrane currents from microglia in acutely isolated cortical mouse brain slices during an experimentally induced CSD. Coincident with the change in the intrinsic optical signal and the negative DC potential shift we recorded an increase in potassium conductance predominantly mediated by K(+) inward rectifier (Kir)2.1, which was blocked by the NMDA receptor antagonist D-AP5. Application of NMDA and an increase in extracellular K(+) mimics the CSD-induced Kir activation. Application of D-AP5, but not the purinergic receptor antagonist RB2, blocks the NMDA-induced Kir activation. The K(+) channel blocker Ba(2+) blocks both the CSD- and the NMDA-triggered increase in Kir channel activity. In addition, we could confirm previous findings that microglia in the adult brain do not express functional NMDA receptors by recording from microglia cultured from adult brain. From these observations we conclude that CSD activates neuronal NMDA receptors, which lead to an increase in extracellular [K(+)] resulting in the activation of Kir channel activity in microglia. Cortical spreading depression (CSD) is a wave of neuronal depolarization spreading through the cortex and is associated with the aura of migraine. Here we show that microglial cells, which are viewed as pathologic sensors of the brain, can sense this wave. The increase in the extracellular potassium concentration associated with that wave leads to the activation of an inward rectifying potassium conductance in microglia. The involvement of neuronal NMDA receptors is crucial because NMDA mimics that response and microglia do not express functional NMDA receptors. Although it is now evident that CSD leads to a signal in microglia, the consequences of this microglial activation during CSD needs to be explored.

The Effect of Variations in Ionic Conductance Values on the Dynamics of a Mathematical Model of Non-Spiking A-Type Horizontal Cells in the Rabbit Retina

A previous study proposed a mathematical model of A-type horizontal cells in the rabbit retina. This model, which was constructed based on the Hodgkin-Huxley model, was described by a system of nonlinear ordinary differential equations. The model contained five types of voltage-dependent ionic conductances: sodium, calcium, delayed rectifier potassium, transient outward potassium, and anomalous rectifier potassium conductances. The previous study indicated that when the delayed rectifier potassium conductance had a small value, depolarizing stimulation could change the dynamic state of the model from a hyperpolarized steady state to a depolarized steady state. However, how this change was affected by variations in the ionic conductance values was not clarified in detail in the previous study. To clarify this issue, in the present study, we performed numerical simulation analysis of the model and revealed the differences among the five types of ionic conductances.

Decrease in an Inwardly Rectifying Potassium Conductance in Mouse Mammary Secretory Cells after Forced Weaning.

Mammary glands are physiologically active in female mammals only during nursing. Immediately after weaning, most lactation-related genes are downregulated and milk production ceases. In our previous study, we have detected an inwardly rectifying potassium channel (Kir) 2.1-like current in mammary secretory (MS) cells freshly isolated from lactating mice. This current is highly sensitive to external Ba2+. The potassium permeability of the Kir channels may contribute to the secretion and/or preservation of ions in milk. We hypothesized that the functions of the Kir channels in MS cells are regulated after weaning. To test this hypothesis, we examined the effect of forced weaning on the Ba2+-sensitive Kir current and Kir2.1 expression in the mouse mammary glands. Twenty-four hours after weaning, the lumina of mammary acini were histologically enlarged by milk accumulation. The whole-cell patch-clamp analyses showed that the Ba2+-sensitive Kir current in the post-weaning MS cells was smaller than in the lactating MS cells. The inward conductances of the current in the lactating and post-weaning cells were 4.25 ± 0.77 and 0.93 ± 0.34 nS, respectively. Furthermore, real-time PCR and Western blot analyses showed that Kir2.1 mRNA and protein expression decreased in the post-weaning mammary gland (mRNA, 90% reduction; protein, 47% reduction). Moreover, the local milk accumulation caused by teat sealing decreased Kir conductance in MS cells (2.74 ± 0.45 and 0.36 ± 0.27 nS for control and sealed mammary glands, respectively). This was concomitant with the reduction in the Kir2.1 mRNA expression. Our results suggest that milk stasis after weaning immediately decreases the Kir conductance in MS cells. This decrease in the Kir conductance may be partly caused by the reduction in the Kir2.1 mRNA and protein expression. These alterations during the post-weaning period may be involved in the cessation of ion secretion and/or preservation in the milk.

μ opioid receptor activation hyperpolarizes respiratory-controlling Kölliker-Fuse neurons and suppresses post-inspiratory drive.

In addition to reductions in respiratory rate, opioids also cause aspiration and difficulty swallowing, indicating impairment of the upper airways. The Kölliker-Fuse (KF) maintains upper airway patency and a normal respiratory pattern. In this study, activation of μ opioid receptors in the KF reduced respiratory frequency and tidal volume in anaesthetized rats. Nerve recordings in an in situ preparation showed that activation of μ opioid receptors in the KF eliminated the post-inspiration phase of the respiratory cycle. In brain slices, μ opioid agonists hyperpolarized a distinct population (61%) of KF neurons by activation of an inwardly rectifying potassium conductance. These results suggest that KF neurons that are hyperpolarized by opioids could contribute to opioid-induced respiratory disturbances, particularly the impairment of upper airways. Opioid-induced respiratory effects include aspiration and difficulty swallowing, suggesting impairment of the upper airways. The pontine Kölliker-Fuse nucleus (KF) controls upper airway patency and regulates respiration, in particular the inspiratory/expiratory phase transition. Given the importance of the KF in coordinating respiratory pattern, the mechanisms of μ opioid receptor activation in this nucleus were investigated at the systems and cellular level. In anaesthetized, vagi-intact rats, injection of opioid agonists DAMGO or [Met(5) ]enkephalin (ME) into the KF reduced respiratory frequency and amplitude. The μ opioid agonist DAMGO applied directly into the KF of the in situ arterially perfused working heart-brainstem preparation of rat resulted in robust apneusis (lengthened low amplitude inspiration due to loss of post-inspiratory drive) that was rapidly reversed by the opioid antagonist naloxone. In brain slice preparations, activation of μ opioid receptors on KF neurons hyperpolarized a distinct population (61%) of neurons. As expected, the opioid-induced hyperpolarization reduced the excitability of the neuron in response to either current injection or local application of glutamate. In voltage-clamp recordings the outward current produced by the opioid agonist ME was concentration dependent, reversed at the potassium equilibrium potential and was blocked by BaCl2 , characteristics of a G protein-coupled inwardly rectifying potassium (GIRK) conductance. The clinically used drug morphine produced an outward current in KF neurons with similar potency to morphine-mediated currents in locus coeruleus brain slice preparations. Thus, the population of KF neurons that are hyperpolarized by μ opioid agonists are likely mediators of the opioid-induced loss of post-inspiration and induction of apneusis.

Nesfatin‐1 influences the excitability of subfornical organ neurons through actions on the Ik conductance (1129.2)

Nesfatin‐1 is produced in several brain areas involved in metabolic processes and has been implicated in the control of ingestive behaviors and cardiovascular function. In this study, we confirm the presence of mRNA for the nucleobindin‐2 gene in the subfornical organ (SFO). We used whole‐cell patch clamp recordings in current clamp configuration to investigate the influence of nesfatin‐1 on the membrane potential of dissociated SFO neurons. We found that 80.3% (49 of 61) of neurons tested showed a response to nesfatin‐1 (100 nM, 10 nM and 1 nM) with 47.5% depolarising and 32.8% hyperpolarising. We then used voltage clamp configuration to investigate the conductances through which nesfatin‐1 may act to change the membrane potential of SFO neurons. Voltage clamp ramp and step protocols revealed a nesfatin‐1 induced enhancement of the delayed rectifier potassium conductance, IK. Pharmacological blockade of this conductance using tetraethyl ammonium chloride (TEA) greatly reduced the magnitude and occurrence of the observed hyperpolarisations, which implies that nesfatin‐1 induced hyperpolarisations are mediated through IK. This study has demonstrated that nesfatin‐1 has the ability to influence the membrane potential of SFO neurons by enhancing the IK conductance, and thus has identified the SFO as a potential site at which nesfatin‐1 may act to regulate ingestive behaviour and cardiovascular control.Grant Funding Source: Canadian Institutes for Health Research

Hydrogen sulfide decreases a slow rectifying potassium conductance in nucleus tractus solitarius neurons (1129.4)

Hydrogen Sulfide (H2S) is an endogenously produced gasotransmitter which has been shown to play a role within the central nervous system. Prior studies have shown that H2S has presynaptic effects on neurons of the nucleus tractus solitarius (NTS) through modulation of glutamate transmission. We have also recently demonstrated postsynaptic effects of H2S on the membrane potential of NTS neurons. The current study was undertaken to investigate the ion channels in NTS neurons modulated by H2S. Using whole cell voltage‐clamp recordings we found that H2S causes a decrease in current evoked in the depolarizing range of a slow voltage ramp indicative of modulation of the voltage activated potassium conductance IK. This conductance was found to decrease by a mean of 20.2 ± 2.5% (n=11) at the end of the ramp (10 mV) following application of 10 mM NaHS, a H2S donor, with all 11 neurons influenced in a similar manner. All cells demonstrated a partial to full recovery of this conductance after wash off of NaHS. The present work suggests that H2S controls the excitability of NTS neurons, at least in part through the modulation of the delayed rectifier potassium conductance, IK.Grant Funding Source: This work was supported by the Canadian Institutes for Health Research.

Cellular Actions of Nesfatin-1 in the Subfornical Organ

Nesfatin-1, a centrally acting anorexigenic peptide, is produced in several brain areas involved in metabolic processes and has been implicated in the control of ingestive behaviours and cardiovascular functions. The present study aimed to determine whether the subfornical organ (SFO), a central nervous system (CNS) site that has been extensively implicated in the regulation of appetite and thirst, may represent a potential site for central actions of nesfatin-1. We first used the reverse transcriptase-polymerase chain reaction and were able to confirm the presence of mRNA for the nucleobindin-2 gene in the SFO. We then used whole-cell patch clamp recordings to investigate the influence of nesfatin-1 on the membrane potential of dissociated SFO neurones. A total of 80.3% (49 of 61) of neurones tested showed a response to nesfatin-1 (100nm, 10nm and 1nm). Of these, 47.5% depolarised, with a mean depolarisation of 8.2±0.9mV (n=29) and 32.8% hyperpolarised with a mean hyperpolarisation of -8.9±1.2mV (n=20). Peak magnitudes were seen at a concentration of 1nm nesfatin-1, whereas no effect was observed at 100pm nesftain-1 (n=3). Furthermore, voltage clamp ramp and step protocols revealed a nesfatin-1 induced activation of the delayed rectifier potassium conductance, IK . Pharmacological blockade of this conductance greatly reduced the magnitude and occurrence of the observed hyperpolarisations. The present study thus demonstrates that nesfatin-1 has the ability to influence the membrane potential of SFO neurones, and thus identifies the SFO as a potential site at which nesfatin-1 may act to regulate ingestive behaviour and cardiovascular control.

Rapid evaluation of a protein-based voltage probe using a field-induced membrane potential change

The development of a high performance protein probe for the measurement of membrane potential will allow elucidation of spatiotemporal regulation of electrical signals within a network of excitable cells. Engineering such a probe requires a functional screen of many candidates. Although the glass-microelectrode technique generally provides an accurate measure of a given test probe, throughputs are limited. In this study, we focused on an approach that uses the membrane potential changes induced by an external electric field in a geometrically simple mammalian cell. For quantitative evaluation of membrane voltage probes that rely on the structural transition of the S1–S4 voltage sensor domain and hence have non-linear voltage dependencies, it was crucial to introduce exogenous inwardly rectifying potassium conductance to reduce cell-to-cell variability in resting membrane potentials. Importantly, the addition of the exogenous conductance drastically altered the profile of the field-induced potential. Following a site-directed random mutagenesis and the rapid screen, we identified a mutant of a voltage probe Mermaid, exhibiting positively shifted voltage sensitivity. Due to its simplicity, the current approach will be applicable under a microfluidic configuration to carry out an efficient screen. Additionally, we demonstrate another interesting aspect of the field-induced optical signals, ability to visualize electrical couplings between cells.

Spontaneous Inhibitory Synaptic Currents Mediated by a G Protein-Coupled Receptor

G protein-coupled receptors (GPCRs) affect many physiological processes by modulating both intrinsic membrane conductances and synaptic transmission. This study describes spontaneous miniature inhibitory postsynaptic currents mediated by vesicular dopamine release acting locally on metabotropic D2 receptors leading to the activation of a G protein-coupled inwardly rectifying potassium conductance. Thus, individual exocytotic events result in spontaneous GPCR-mediated transmission, similar to synaptic activation of classical ligand-gated ion channels.

Apelin acts in the subfornical organ to influence neuronal excitability and cardiovascular function

Apelin is an adipocyte-derived hormone involved in the regulation of water balance, food intake and the cardiovascular system partially through actions in the CNS. The subfornical organ (SFO) is a circumventricular organ with identified roles in body fluid homeostasis, cardiovascular control and energy balance. The SFO lacks a normal blood-brain barrier, and is thus able to detect circulating signalling molecules such as angiotensin II and leptin. In this study, we investigated actions of apelin-13, the predominant apelin isoform in brain and circulatory system, on the excitability of dissociated SFO neurons using electrophysiological approaches, and determined the cardiovascular consequences of direct administration into the SFO of anaesthetized rats. Whole cell current clamp recording revealed that bath-applied 100 nm apelin-13 directly influences the excitability of the majority of SFO neurons by eliciting either depolarizing (31.8%, mean 7.0 ± 0.8 mV) or hyperpolarizing (28.6%, mean -10.4 ± 1.8 mV) responses. Using voltage-clamp techniques, we also identified modulatory actions of apelin-13 on specific ion channels, demonstrating that apelin-13 activates a non-selective cationic conductance to depolarize SFO neurons while activation of the delayed rectifier potassium conductance underlies hyperpolarizing effects. In anaesthetized rats, microinjection of apelin into SFO decreased both blood pressure (BP) (mean area under the curve -1492.3 ± 357.1 mmHg.s, n = 5) and heart rate (HR) (-32.4 ± 10.39 beats, n = 5). Our data suggest that circulating apelin can directly affect BP and HR as a consequence of the ability of this peptide to modulate the excitability of SFO neurons.

Selective Silencing of Individual Dendritic Branches by an mGlu2-Activated Potassium Conductance in Dentate Gyrus Granule Cells

Group II metabotropic glutamate receptors (mGlu-IIs) modulate hippocampal information processing through several presynaptic actions. We describe a novel postsynaptic inhibitory mechanism mediated by the mGlu2 subtype that activates an inwardly rectifying potassium conductance in the dendrites of DG granule cells of rats and mice. Data from glutamate-uncaging experiments and simulations indicate that mGlu2-activated potassium conductance uniformly reduces the peak amplitude of synaptic inputs arriving in the distal two-thirds of dendrites, with only minor effects on proximal inputs. This unique shunting profile is consistent with a peak expression of the mGlu2-activated conductance at the transition between the proximal and middle third of the dendrites. Further simulations under various physiologically relevant conditions showed that when a shunting conductance was activated in the proximal third of a single dendrite, it effectively modulated input to this specific branch while leaving inputs in neighboring dendrites relatively unaffected. Therefore, the restricted expression of the mGlu2-activated potassium conductance in the proximal third of DG granule cell dendrites represents an optimal localization for achieving the opposing biophysical requirements for uniform yet selective modulation of individual dendritic branches.

Inactivating ion channels augment robustness of subthreshold intrinsic response dynamics to parametric variability in hippocampal model neurons

Voltage-gated ion channels play a critical role in regulating neuronal intrinsic response dynamics (IRD). Here, we computationally analysed the roles of the two inactivating subthreshold conductances (A and T), individually and in various combinations with the non-inactivating h conductance, in regulating several physiological IRD measurements in the theta frequency range. We found that the independent presence of a T conductance, unlike that of an h conductance, was unable to sustain an inductive phase lead in the theta frequency range, despite its ability to mediate theta frequency resonance. The A conductance, on the other hand, when expressed independently, acted in a manner similar to a leak conductance with reference to most IRD measurements. Next, analysing the impact of pair-wise coexpression of these channels, we found that the coexpression of the h and T conductances augmented the range of parameters over which they sustained resonance and inductive phase lead. Additionally, coexpression of the A conductance with the h or the T conductance elicited changes in IRD measurements that were similar to those obtained with the expression of a leak conductance with a resonating conductance. Finally, to understand the global sensitivity of IRD measurements to all parameters associated with models expressing all three channels, we generated 100,000 neuronal models, each built with a unique set of parametric values. We categorized valid models among these by matching their IRD measurements with experimental counterparts, and found that functionally similar models could be achieved even when underlying parameters displayed tremendous variability and exhibited weak pair-wise correlations. Our results suggest that the three prominent subthreshold conductances contribute differently to intrinsic excitability and to phase coding. We postulate that the differential expression and activity-dependent plasticity of these conductances contribute to robustness of subthreshold IRD, whereby response homeostasis is achieved by recruiting several non-unique combinations of these channel parameters.

Morphine Desensitization and Cellular Tolerance Are Distinguished in Rat Locus Ceruleus Neurons

μ-Opioid receptor desensitization is considered an initial step in the development of tolerance. Curiously, the commonly used opioid morphine produces robust tolerance but minimal acute desensitization. This study was designed to test the hypothesis that desensitization is indeed present in morphine-treated animals and is distinguished from cellular tolerance by time course of recovery and mechanism. To induce tolerance, rats were treated with continuously released morphine for 1 week. Morphine-mediated activation of G protein-coupled inwardly rectifying potassium conductance was measured using voltage-clamp recordings from locus ceruleus neurons in brain slices from naive or morphine-treated rats. Cellular tolerance was observed as a decrease in morphine efficacy in slices from morphine-treated rats. This tolerance persisted for at least 6 h. An additional reduction in morphine-mediated current was observed when slices from morphine-treated rats were continuously maintained in morphine at approximately the circulating plasma concentration. This additional reduction recovered within 1 h after removal of morphine from the slice and represents desensitization that developed in the tolerant animal. Recovery from desensitization, but not long-lasting tolerance, was facilitated by protein phosphatase 1 (PP1) activity. Furthermore, desensitization, but not tolerance, was reversed by protein kinase C (PKC) inhibitor but not by an inhibitor of c-Jun N-terminal kinase. Therefore, morphine treatment leads to both long-lasting cellular tolerance and readily reversible desensitization, which are differentially dependent on PP1 and PKC activity and combine to result in a substantial decrease in morphine effectiveness. This PKC-mediated desensitization may contribute to the previously reported PKC-dependent reversal of behavioral tolerance.