Role Of Voltage-gated Ion Channels Research Articles

A modelling study to dissect the potential role of voltage-gated ion channels in activity-dependent conduction velocity changes as identified in small fiber neuropathy patients.

Patients with small fiber neuropathy (SFN) suffer from neuropathic pain, which is still a therapeutic problem. Changed activation patterns of mechano-insensitive peripheral nerve fibers (CMi) could cause neuropathic pain. However, there is sparse knowledge about mechanisms leading to CMi dysfunction since it is difficult to dissect specific molecular mechanisms in humans. We used an in-silico model to elucidate molecular causes of CMi dysfunction as observed in single nerve fiber recordings (microneurography) of SFN patients. We analyzed microneurography data from 97 CMi-fibers from healthy individuals and 34 of SFN patients to identify activity-dependent changes in conduction velocity. Using the NEURON environment, we adapted a biophysical realistic preexisting CMi-fiber model with ion channels described by Hodgkin-Huxley dynamics for identifying molecular mechanisms leading to those changes. Via a grid search optimization, we assessed the interplay between different ion channels, Na-K-pump, and resting membrane potential. Changing a single ion channel conductance, Na-K-pump or membrane potential individually is not sufficient to reproduce in-silico CMi-fiber dysfunction of unchanged activity-dependent conduction velocity slowing and quicker normalization of conduction velocity after stimulation as observed in microneurography. We identified the best combination of mechanisms: increased conductance of potassium delayed-rectifier and decreased conductance of Na-K-pump and depolarized membrane potential. When the membrane potential is unchanged, opposite changes in Na-K-pump and ion channels generate the same effect. Our study suggests that not one single mechanism accounts for pain-relevant changes in CMi-fibers, but a combination of mechanisms. A depolarized membrane potential, as previously observed in patients with neuropathic pain, leads to changes in the contribution of ion channels and the Na-K-pump. Thus, when searching for targets for the treatment of neuropathic pain, combinations of several molecules in interplay with the membrane potential should be regarded.

Electrical stimulation enhances the neuronal differentiation of neural stem cells in three-dimensional conductive scaffolds through the voltage-gated calcium ion channel

Electrical stimulation (ES) or electroconductive scaffold has been proved to have the positive effects on the behavior of neural stem cells (NSCs). We previously developed a novel three-dimensional conductive composite scaffold of poly (3, 4-ethylenedioxythiophen)/chitosan/gelatin (PEDOT/Cs/Gel) for neural tissue engineering. In the present study, we further studied the effect of three-dimensional conductive scaffolds combined with ES on the neuronal differentiation of NSCs. The sandwiched ES device was designed to apply single-phase pulse voltage on NSCs cultured in conductive scaffold for 7 days (4 h/day). Proliferation and differentiation related proteins and genes were analyzed by immunofluorescence staining and RT-qPCR. The role of voltage-gated ion channels (VGICs) in regulating NSCs’ neuronal differentiation by ES was investigated in presence of ion channels blockers. The results of protein and gene expression indicated that ES not only promoted the proliferation of NSCs cultured in the conductive scaffold, but also enhanced the differentiation of NSCs into neurons. Especially, the voltage-gated calcium channel (Cav2+) played an important role in the neuronal differentiation of NSCs under ES. Our findings demonstrated that ES combined with three-dimensional conductive scaffolds would be a promising strategy to regulate the neuronal differentiation of NSCs for neural regeneration.

The role of voltage-gated ion channels in visual function and disease in mammalian photoreceptors.

Light activation of the classical light-sensing retinal neurons, the photoreceptors, results in a graded change in membrane potential that ultimately leads to a reduction in neurotransmitter release to the post-synaptic retinal neurons. Photoreceptors show striking powers of adaptation, and for visual processing to function optimally, they must adjust their gain to remain responsive to different levels of ambient light intensity. The presence of a tightly controlled balance of inward and outward currents modulated by several different types of ion channels is what gives photoreceptors their remarkably dynamic operating range. Part of the resetting and modulation of this operating range is controlled by potassium and calcium voltage-gated channels, which are involved in setting the dark resting potential and synapse signal processing, respectively. Their essential contribution to visual processing is further confirmed in patients suffering from cone dystrophy with supernormal rod response (CDSRR) and congenital stationary night blindness type 2 (CSNB2), both conditions that lead to irreversible vision loss. This review will discuss these two types of voltage-gated ion channels present in photoreceptors, focussing on their structure and physiology, and their role in visual processing. It will also discuss the use and benefits of knockout mouse models to further study the function of these channels and what routes to potential treatments could be applied for CDSRR and CSNB2.

Effects of electromagnetic fields on neuronal ion channels: a systematic review.

Many aspects of chemistry and biology are mediated by electromagnetic field (EMF) interactions. The central nervous system (CNS) is particularly sensitive to EMF stimuli. Studies have explored the direct effect of different EMFs on the electrical properties of neurons in the last two decades, particularly focusing on the role of voltage-gated ion channels (VGCs). This work aims to systematically review published evidence in the last two decades detailing the effects of EMFs on neuronal ion channels as per the PRISM guidelines. Following a predetermined exclusion and inclusion criteria, 22 papers were included after searches on three online databases. Changes in calcium homeostasis, attributable to the voltage-gated calcium channels, were found to be the most commonly reported result of EMF exposure. EMF effects on the neuronal landscape appear to be diverse and greatly dependent on parameters, such as the field's frequency, exposure time, and intrinsic properties of the irradiated tissue, such as the expression of VGCs. Here, we systematically clarify how neuronal ion channels are particularly affected and differentially modulated by EMFs at multiple levels, such as gating dynamics, ion conductance, concentration in the membrane, and gene and protein expression. Ion channels represent a major transducer for EMF-related effects on the CNS.

Examining Sodium and Potassium Channel Conductances Involved in Hyperexcitability of Chemotherapy-Induced Peripheral Neuropathy: A Mathematical and Cell Culture-Based Study.

Chemotherapy-induced peripheral neuropathy (CIPN) is a prevalent, painful side effect which arises due to a number of chemotherapy agents. CIPN can have a prolonged effect on quality of life. Chemotherapy treatment is often reduced or stopped altogether because of the severe pain. Currently, there are no FDA-approved treatments for CIPN partially due to its complex pathogenesis in multiple pathways involving a variety of channels, specifically, voltage-gated ion channels. One aspect of neuropathic pain in vitro is hyperexcitability in dorsal root ganglia (DRG) peripheral sensory neurons. Our study employs bifurcation theory to investigate the role of voltage-gated ion channels in inducing hyperexcitability as a consequence of spontaneous firing due to the common chemotherapy agent paclitaxel. Our mathematical investigation of a reductionist DRG neuron model comprised of sodium channel Nav1.7, sodium channel Nav1.8, delayed rectifier potassium channel, A-type transient potassium channel, and a leak channel suggests that Nav1.8 and delayed rectifier potassium channel conductances are critical for hyperexcitability of small DRG neurons. Introducing paclitaxel into the model, our bifurcation analysis predicts that hyperexcitability is highest for a medium dose of paclitaxel, which is supported by multi-electrode array (MEA) recordings. Furthermore, our findings using MEA reveal that Nav1.8 blocker A-803467 and delayed rectifier potassium enhancer L-alpha-phosphatidyl-D-myo-inositol 4,5-diphosphate, dioctanoyl (PIP2) can reduce paclitaxel-induced hyperexcitability of DRG neurons. Our approach can be readily extended and used to investigate various other contributors of hyperexcitability in CIPN.

Electrical stimulation of adipose-derived mesenchymal stem cells in conductive scaffolds and the roles of voltage-gated ion channels

Since electrical stimulation (ES) can significantly accelerate bone healing, a conductive scaffold that can deliver ES locally at the defect site is desirable for bone defect therapy. Herein, an electrically conductive scaffold was prepared via incorporation of polypyrrole (PPY) in a polycaprolactone (PCL) template scaffold. In vitro tests with mouse osteoblasts indicate that the PPY/PCL scaffold has good biocompatibility, and is suitable for use as an ES substrate. When human adipose-derived mesenchymal stem cells (AD-MSCs) were cultured in the PPY/PCL scaffold and subjected to 200μA of direct current for 4h per day for 21days, the amount of calcium deposited was 100% higher than that without ES. When these cells were subjected to ES together with blockers of voltage-gated calcium (Ca2+v), sodium (Na+v), potassium (K+v), or chloride (Cl−v) channels, the ES-induced enhancement of AD-MSCs’ functions was reduced with Na+v, K+v, or Cl−v blockers and completely nullified with Ca2+v blocker. These results indicate that ion fluxes through these channels activated by ES induce different cascades of reactions in the cells, which subsequently regulate AD-MSCs’ functions, and Ca2+v plays a more critical role than the other three channels. Our results further the current understanding of the mechanisms by which ES regulates stem cells’ behavior, and also showed that the conductive PPY/PCL scaffold with application of ES has good potential in bone defect therapy. Statement of SignificanceIn this work, an electrically conductive and biocompatible scaffold was prepared by incorporating polypyrrole in a polycaprolactone template scaffold. Application of 200μA direct current for 4h per day to human adipose derived-mesenchymal stem cells cultured on this scaffold promoted migration of these cells into the inner region of the scaffold and enhanced their osteogenic differentiation. The roles of voltage-gated ion channels (Ca2+v, Na+v, K+v and Cl−v) in osteogenic differentiation stimulated by the electric current were investigated. The results from these experiments further the current understanding of the mechanisms by which electrical stimulation regulates stem cells’ behavior, and also show that the polypyrrole–polycaprolactone scaffold with application of electrical stimulation has good potential in bone defect therapy.

Channelopathies and dendritic dysfunction in fragile X syndrome

Dendritic spine abnormalities and the metabotropic glutamate receptor theory put the focus squarely on synapses and protein synthesis as the cellular locus of fragile X syndrome. Synapses however, are only partly responsible for information processing in neuronal networks. Neurotransmitter triggered excitatory postsynaptic potentials (EPSPs) are shaped and integrated by dendritic voltage-gated ion channels. These EPSPs, and in some cases the resultant dendritic spikes, are further modified by dendritic voltage-gated ion channels as they propagate to the soma. If the resultant somatic depolarization is large enough, action potential(s) will be triggered and propagate both orthodromically down the axon, where it may trigger neurotransmitter release, and antidromically back into the dendritic tree, where it can activate and modify dendritic voltage-gated and receptor activated ion channels. Several channelopathies, both soma-dendritic (L-type calcium channels, Slack potassium channels, h-channels, A-type potassium channels) and axo-somatic (BK channels and delayed rectifier potassium channels) were identified in the fmr1-/y mouse model of fragile X syndrome. Pathological function of these channels will strongly influence the excitability of individual neurons as well as overall network function. In this chapter we discuss the role of voltage-gated ion channels in neuronal processing and describe how identified channelopathies in models of fragile X syndrome may play a role in dendritic pathophysiology.

Activity-Dependent Control of Neuronal Output by Local and Global Dendritic Spike Attenuation

Neurons possess elaborate dendritic arbors which receive and integrate excitatory synaptic signals. Individual dendritic subbranches exhibit local membrane potential supralinearities, termed dendritic spikes, which control transfer of local synaptic input to the soma. Here, we show that dendritic spikes in CA1 pyramidal cells are strongly regulated by specific types of prior input. While input in the linear range is without effect, supralinear input inhibits subsequent spikes, causing them to attenuate and ultimately fail due to dendritic Na(+) channel inactivation. This mechanism acts locally within the boundaries of the input branch. If an input is sufficiently strong to trigger axonal action potentials, their back-propagation into the dendritic tree causes a widespread global reduction in dendritic excitability which is prominent after firing patterns occurring in vivo. Together, these mechanisms control the capability of individual dendritic branches to trigger somatic action potential output. They are invoked at frequencies encountered during learning, and impose limits on the storage and retrieval rates of information encoded as branch excitability.

Voltage-clamp analysis and computational model of dopaminergic neurons from mouse retina

Isolated dopaminergic amacrine (DA) cells in mouse retina fire rhythmic, spontaneous action potentials and respond to depolarizing current with trains of low-frequency action potentials. To investigate the roles of voltage-gated ion channels in these processes, the transient A-type K+ current (I(K,A)) and Ca2+ current (I(Ca)) in isolated mouse DA cells were analyzed by voltage clamp. The I(K,A) activated at -60 mV and inactivated rapidly. I(Ca) activated at around -30 mV and reached a peak at 10 mV without apparent inactivation. We also extended our previous computational model of the mouse DA cell to include the new electrophysiological data. The model consisted of a membrane capacitance in parallel with eight currents: Na+ transient (I(Na,T)), Na+ persistent (I(Na,P)), delayed rectifier potassium (I(Kdr)), I(K,A), calcium-dependent potassium (I(K,Ca)), L-type Ca2+ I(Ca), hyperpolarization-activated cation current (I(h)), and a leak current (I(L)). Hodgkin-Huxley type equations were used to define the voltage- and time-dependent activation and inactivation. The simulations were implemented using the neurosimulator SNNAP. The model DA cell was spontaneously active from a wide range of initial membrane potentials. The spontaneous action potentials reached 35 mV at the peak and hyperpolarized to -76 mV between spikes. The spontaneous firing frequency in the model was 6 Hz. The model DA cell responded to prolonged depolarizing current injection by increasing its spiking frequency and eventually reaching a depolarization block at membrane potentials greater than -10 mV. The most important current for determining the firing rate was I(K,A). When the amplitude of I(K,A) was decreased, the firing rate increased. I(Ca) and I(K,Ca) also affected the width of action potentials but had only minor effects on the firing rate. Ih affected the firing rate slightly but did not change the waveform of the action potentials.

Controversy surrounding the existence of discrete functional classes of astrocytes in adult gray matter.

Since 1992, it has been possible to record ionic currents from identified astrocytes in situ, using brain slice technology. Brain slice recordings confirm previous in vitro findings that expression of voltage-gated K(+) and Na(+) channels are a feature of this cell type. In contrast to cultured astrocytes, most investigators found that astrocytes in situ did not contain detectable, or at very best only low, levels of glial fibrillary acidic protein (GFAP). Structural and immunocytochemical investigations determined that these cells are different from oligodendrocyte precursors. In addition to cells with this current pattern, many but not all investigators found a second pool of astrocytes with no voltage-gated ion channels and high GFAP content. These two subpopulations of cells were termed complex and passive astrocytes. The existence of passive astrocytes has been questioned because of possible problems with space clamp conditions and spillage of EGTA-buffered pipette solution around the cells before recordings. Another problem is the fact there is a discrepancy regarding the GFAP content of complex astrocytes. It is of interest that recent immunocytochemical studies suggest the existence of two pools of astrocytes, one with a high GFAP content and one with nondetectable GFAP. Given this, it is tempting to correlate the two (controversial) electrophysiological patterns with immunochemical differences (GFAP) in order to demonstrate two functionally discrete classes of astrocytes in adult gray matter. However, despite evidence that some of the K(+) channels may be involved in proliferation, the role of voltage-gated ion channels in this nonexcitable cell type remains unknown. This is despite the fact that astrocytic Na(+) channels show dramatic changes after pathological events, re-enforcing the notion that the expression of this channel is under tight neuronal control. Several studies suggest that there is a great degree of flexibility and that astrocytes can undergo rapid changes in expression of both membrane ion currents and GFAP. Although it is likely that astrocytes exhibit different structural and membrane properties, this heterogeneity might be a reflection of the flexible plasticity of one astrocyte type under influence of environmental factors rather than of the existence of two distinct and permanent subtypes.

Cerebral function before and after extra-intracranial carotid bypass.

Thirty-eight patients had their cerebral function measured before and after extra-intracranial carotid bypass surgery using the Halstead-Reitan neuropsychology test battery. Two composite indices of cerebral function for each patient showed them to be impaired before operation. There was no improvement in the composite measures after operation to match that previously demonstrated after carotid endarterectomy. This may reflect the greater pre-operative cerebral impairment of the extra-intracranial group.