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Cutaneous thermal hyperemia during local skin heating to 39°C, which is highly dependent on nitric oxide synthase (NOS), is frequently used to assess endothelium-dependent cutaneous vasodilation. We showed that Na+-K+-ATPase mediates the regulation of cutaneous thermal hyperemia partly via NOS-dependent mechanisms although a component of the Na+-K+-ATPase modulation of cutaneous thermal hyperemia is NOS independent. Thus, as with NOS, Na+-K+-ATPase may be important in the regulation of cutaneous endothelial vascular function.

In myelinated nerve fibers, action potentials are generated at nodes of Ranvier. These structures are located at interruptions of the myelin sheath, forming narrow gaps with small rings of axolemma freely exposed to the extracellular space. The mammalian node contains a high density of Na channels and K-selective leakage channels. Voltage-dependent Kv1 channels are only present in the juxta-paranode. Recently, the leakage channels have been identified as K2P channels (TRAAK, TREK-1). K2P channels are K-selective "background" channels, characterized by outward rectification and their ability to be activated, e.g. by temperature, mechanical stretch, or arachidonic acid. We are only beginning to elucidate the peculiar functions of nodal K2P channels. I will discuss two functions of the nodal K2P-mediated conductance. First, at body temperature K2P channels have a high open probability, thereby inducing a resting potential of about -85 mV. This negative resting potential reduces steady-state Na channel inactivation and ensures a large Na inward current upon a depolarizing stimulus. Second, the involvement of the K2P conductance in nodal action potential repolarization. The identification of nodal K2P channels is exciting since it shows that the nodal K conductance is not a fixed value but can be changed, it can be increased or decreased by a broad range of K2P modulators, thereby modulating e.g. the resting potential. I will exemplify the functional importance of nodal K2P channels by describing in more detail the function of the K2P conductance increase by increasing the temperature to 37°C. This article is protected by copyright. All rights reserved.

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.

A model of activity generation in the epileptic focus was proposed based on the ideas that synchronization occurs between the oscillations that arise to regulate the resting potential level and that neuronal discharges are synchronized with slow oscillations. Four phases were identified in seizure development: an increase in slow-wave activity, high-frequency spike activity, synchronization in the presence of sharp peaks, and extinction of the seizure. The model was compared with an intraoperative electrocorticography record.

The nonpsychoactive phytocannabinoid cannabidiol (CBD) has been shown to have analgesic effects in animal studies but little is known about its mechanism of action. We examined the effects of CBD on intrinsic excitability of primary pain-sensing neurons. Studying acutely dissociated capsaicin-sensitive mouse DRG neurons at 37°C, we found that CBD effectively inhibited repetitive action potential firing, from 15–20 action potentials evoked by 1 s current injections in control to 1–3 action potentials with 2 μm CBD. Reduction of repetitive firing was accompanied by a reduction of action potential height, widening of action potentials, reduction of the afterhyperpolarization, and increased propensity to enter depolarization block. Voltage-clamp experiments showed that CBD inhibited both TTX-sensitive and TTX-resistant (TTX-R) sodium currents in a use-dependent manner. CBD showed strong state-dependent inhibition of TTX-R channels, with fast binding to inactivated channels during depolarizations and slow unbinding on repolarization. CBD alteration of channel availability at various voltages suggested that CBD binds especially tightly [Kd (dissociation constant), ∼150 nm] to the slow inactivated state of TTX-R channels, which can be substantially occupied at voltages as negative as −40 mV. Remarkably, CBD was more potent in inhibiting TTX-R channels and inhibiting action potential firing than the local anesthetic bupivacaine. We conclude that CBD might produce some of its analgesic effects by direct effects on neuronal excitability, with tight binding to the slow inactivated state of Nav1.8 channels contributing to effective inhibition of repetitive firing by modest depolarizations. SIGNIFICANCE STATEMENT Cannabidiol (CBD) has been shown to inhibit pain in various rodent models, but the mechanism of this effect is unknown. We describe the ability of CBD to inhibit repetitive action potential firing in primary nociceptive neurons from mouse dorsal root ganglia and analyze the effects on voltage-dependent sodium channels. We find that CBD interacts with TTX-resistant sodium channels in a state-dependent manner suggesting particularly tight binding to slow inactivated states of Nav1.8 channels, which dominate the overall inactivation of Nav1.8 channels for small maintained depolarizations from the resting potential. The results suggest that CBD can exert analgesic effects in part by directly inhibiting repetitive firing of primary nociceptors and suggest a strategy of identifying compounds that bind selectively to slow inactivated states of Nav1.8 channels for developing effective analgesics.

Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) are supposed to be a good human-based model, with virtually unlimited cell source, for studies on mechanisms underlying cardiac development and cardiac diseases, and for identification of drug targets. However, a major drawback of hPSC-CMs as a model system, especially for electrophysiological studies, is their depolarized state and associated spontaneous electrical activity. Various approaches are used to overcome this drawback, including the injection of "synthetic" inward rectifier potassium current (IK1), which is computed in real time, based on the recorded membrane potential ("dynamic clamp"). Such injection of an IK1-like current results in quiescent hPSC-CMs with a nondepolarized resting potential that show "adult-like" action potentials on stimulation, with functional availability of the most important ion channels involved in cardiac electrophysiology. These days, dynamic clamp has become a widely appreciated electrophysiological tool. However, setting up a dynamic clamp system can still be laborious and difficult, both because of the required hardware and the implementation of the dedicated software. In the present review, we first summarize the potential mechanisms underlying the depolarized state of hPSC-CMs and the functional consequences of this depolarized state. Next, we explain how an existing manual patch clamp setup can be extended with dynamic clamp. Finally, we shortly validate the extended setup with atrial-like and ventricular-like hPSC-CMs. We feel that dynamic clamp is a highly valuable tool in the field of cellular electrophysiological studies on hPSC-CMs and hope that our directions for setting up such dynamic clamp system may prove helpful.

Background and purpose Sodium channel inhibitors can be used to treat hyperexcitability-related diseases, including epilepsies, pain syndromes, neuromuscular disorders and cardiac arrhythmias. The applicability of these drugs is limited by their nonspecific effect on physiological function. They act mainly by sodium channel block and in addition by modulation of channel kinetics. While channel block inhibits healthy and pathological tissue equally, modulation can preferentially inhibit pathological activity. An ideal drug designed to target the sodium channels of pathological tissue would act predominantly by modulation. Thus far, no such drug has been described. Experimental approach Patch-clamp experiments with ultra-fast solution exchange and photolabeling-coupled electrophysiology were applied to describe the unique mechanism of riluzole on Nav1.4 sodium channels. In silico docking experiments were used to study the molecular details of binding. Key results We present evidence that riluzole acts predominantly by non-blocking modulation. We propose that, being a relatively small molecule, riluzole is able to stay bound to the binding site, but nonetheless stay off the conduction pathway, by residing in one of the fenestrations. We demonstrate how this mechanism can be recognized. Conclusions and implications Our results identify riluzole as the prototype of this new class of sodium channel inhibitors. Drugs of this class are expected to selectively prevent hyperexcitability, while having minimal effect on cells firing at a normal rate from a normal resting potential.

Key points Cartwheel neurons provide potent inhibition to fusiform neurons in the dorsal cochlear nucleus (DCN). Most cartwheel neurons fire action potentials spontaneously, but the ion channels responsible for this intrinsic activity are unknown. We investigated the ion channels responsible for the intrinsic firing of cartwheel neurons and the stable resting membrane potential found in a fraction of these neurons (quiet neurons). Among the ion channels controlling membrane potential of cartwheel neurons, the presence of open ATP-sensitive potassium channels (KATP ) is responsible for the existence of quiet neurons. Our results pinpoint KATP channel modulation as a critical factor controlling the firing of cartwheel neurons. Hence, it is a crucial channel influencing the balance of excitation and inhibition in the DCN. Abstract Cartwheel neurons from the dorsal cochlear nucleus (DCN) are glycinergic interneurons and the primary source of inhibition on the fusiform neurons, the DCN's principal excitatory neuron. Most cartwheel neurons present spontaneous firing (active neurons), producing a steady inhibitory tone on fusiform neurons. In contrast, a small fraction of these neurons do not fire spontaneously (quiet neurons). Hyperactivity of fusiform neurons is seen in animals with behavioural evidence of tinnitus. Because of its relevance in controlling the excitability of fusiform neurons, we investigated the ion channels responsible for the spontaneous firing of cartwheel neurons in DCN slices from rats. We found that quiet neurons presented an outward conductance not seen in active neurons, which generates a stable resting potential. This current was sensitive to tolbutamide, an ATP-sensitive potassium channel (KATP ) antagonist. After inhibition with tolbutamide, quiet neurons start to fire spontaneously, while the active neurons were not affected. On the other hand, in active neurons, KATP agonist diazoxide activated a conductance similar to quiet neurons' KATP conductance and stopped spontaneous firing. According to the effect of KATP channels on cartwheel neuron firing, glycinergic neurotransmission in DCN was increased by tolbutamide and decreased by diazoxide. Our results reveal a role of KATP channels in controlling the spontaneous firing of neurons not involved in fuel homeostasis.

BACKGROUND/AIMS: Tea, produced from the evergreen Camellia sinensis, has reported therapeutic properties against multiple pathologies, including hypertension. Although some studies validate the health benefits of tea, few have investigated the molecular mechanisms of action. The KCNQ5 voltage-gated potassium channel contributes to vascular smooth muscle tone and neuronal M-current regulation.METHODS: We applied electrophysiology, myography, mass spectrometry and in silico docking to determine effects and their underlying molecular mechanisms of tea and its components on KCNQ channels and arterial tone.RESULTS: A 1% green tea extract (GTE) hyperpolarized cells by augmenting KCNQ5 activity >20-fold at resting potential; similar effects of black tea were inhibited by milk. In contrast, GTE had lesser effects on KCNQ2/Q3 and inhibited KCNQ1/E1. Tea polyphenols epicatechin gallate (ECG) and epigallocatechin-3-gallate (EGCG), but not epicatechin or epigallocatechin, isoform-selectively hyperpolarized KCNQ5 activation voltage dependence. In silico docking and mutagenesis revealed that activation by ECG requires KCNQ5-R212, at the voltage sensor foot. Strikingly, ECG and EGCG but not epicatechin KCNQ-dependently relaxed rat mesenteric arteries.CONCLUSION: KCNQ5 activation contributes to vasodilation by tea; ECG and EGCG are candidates for future anti-hypertensive drug development.

Electromyography is a neurophysiological technique for examining the electrical activity of skeletal muscles. It occurred from the muscle membrane potential. In a healthy muscle at rest, the spontaneous physiological activity can be a record by End plate potential, insertion activity. EMG recording of the activity of selective cranial nerve muscles is currently included in the intraoperative set during surgical manipulation of the brainstem. When during the process, weak muscles can be detected and treat those muscles to regain their function. EMG is also used to diagnose neuromuscular performance. Its gain is less and frequency response may be broader. Using the EMG technique to record the potential of muscle is an easy way to generate the resting potential of muscle.

Background The toxicity of silica nanoparticles (SiNPs) on cardiac electrophysiology has seldom been evaluated. Methods Patch-clamp was used to investigate the acute effects of SiNP-100 (100 nm) and SiNP-20 (20 nm) on the transmembrane potentials (TMPs) and ion channels in cultured neonatal mouse ventricular myocytes. Calcium mobilization in vitro, cardiomyocyte ROS generation, and LDH leakage after exposure to SiNPs in vitro and in vivo were measured using a microplate reader. Surface electrocardiograms were recorded in adult mice to evaluate the arrhythmogenic effects of SiNPs in vivo. SiNP endocytosis was observed using transmission electron microscopy. Results Within 30 min, both SiNPs (10-8-10-6 g/mL) did not affect the resting potential and IK1 channels. SiNP-100 increased the action potential amplitude (APA) and the INa current density, but SiNP-20 decreased APA and INa density. SiNP-100 prolonged the action potential duration (APD) and decreased the Ito current density, while SiNP-20 prolonged or shortened the APD, depending on exposure concentrations and increased Ito density. Both SiNPs (10-6 g/mL) induced calcium mobilization but did not increase ROS and LDH levels and were not endocytosed within 10 min in cardiomyocytes in vitro. In vivo, SiNP-100 (4-10 mg/kg) and SiNP-20 (4-30 mg/kg) did not elevate myocardial ROS but increased LDH levels depending on dose and exposure time. The same higher dose of SiNPs (intravenously injected) induced tachyarrhythmias and lethal bradyarrhythmias within 90 min in adult mice. Conclusion SiNPs (i) exert rapid toxic effects on the TMPs of cardiomyocytes in vitro largely owing to their direct interfering effects on the INa and Ito channels and Ca2+ homeostasis but not IK1 channels and ROS levels, and (ii) induce tachyarrhythmias and lethal bradyarrhythmias in vivo. SiNP-100 is more toxic than SiNP-20 on cardiac electrophysiology, and the toxicity mechanism is likely more complicated in vivo.

Illuminated giant cells of characean algae exhibit membrane excitability as well as the spatial patterns of photosynthesis and transmembrane H+ fluxes. The excitation of plasmalemma under these conditions results in the transient degradation of external alkaline and acid zones and inhibits photosynthesis in the alkaline zones. The generation of action potential in the patterned internodes is followed by cell hyperpolarization that peaks in 1 min and lasts up to 15 min. In order to exclude the influence of drifting resting potential on the chloroplast response to plasma membrane excitation, the voltage clamp mode was applied in this work, and chlorophyll fluorescence changes caused by a short depolarizing pulse were monitored. The depolarizing shift of membrane potential under voltage clamp conditions was found to induce a large depression of $$F_{{\text{m}}}^{{{'}}}$$ chlorophyll fluorescence and photosynthetic activity, provided that inward Ca2+ and Cl– currents were triggered and that a steady-state inward H+ flux (or OH– efflux) persisted before the application of an electric stimulus. The depolarization-induced ion currents measured in the alkaline and acidic cell regions under light and in darkness were found to differ significantly. The results are consistent with the notion that the massive inward H+ flow occurring in the alkaline cell regions under illumination is associated with the acidic shift of cytoplasmic pH. Divergent amplitudes of ionic currents in different cell parts can be partially determined by the presence of numerous plasmalemmal invaginations, charasomes specifically localized in the acid zones, as well as by sharp local changes in external pH in acid zones during the perforation of cell wall with a measuring microelectrode.

Background: Although the chondrocyte is a nonexcitable cell, there is strong interest in gaining detailed knowledge of its ion pumps, channels, exchangers, and transporters. In combination, these transport mechanisms set the resting potential, regulate cell volume, and strongly modulate responses of the chondrocyte to endocrine agents and physicochemical alterations in the surrounding extracellular microenvironment. Materials and Methods: Mathematical modeling was used to assess the functional roles of energy-requiring active transport, the Na+/K+ pump, in chondrocytes. Results: Our findings illustrate plausible physiological roles for the Na+/K+ pump in regulating the resting membrane potential and suggest ways in which specific molecular components of pump can respond to the unique electrochemical environment of the chondrocyte. Conclusion: This analysis provides a basis for linking chondrocyte electrophysiology to metabolism and yields insights into novel ways of manipulating or regulating responsiveness to external stimuli both under baseline conditions and in chronic diseases such as osteoarthritis.

All cells maintain a voltage across their plasma membranes. Only excitable cells, however, can generate action potentials, the rapid, transient changes in membrane potential that spread along the surface of these unique cells. Action potential generation and propagation occurs through, and is regulated by, the function of voltage-gated ion channels – proteins with ion-selective pores that span the cell membrane. Ion channels undergo changes in their structural conformation in response to changes in the electrical field across the membrane. These structural changes cause the opening of pores – channels – through which ions can flow down their electrochemical gradient. The charge carried by ions creates an electrical current and rapidly alters the membrane potential with time- and voltage-dependent properties. This rapid, transient membrane potential change is called the action potential. Action potentials transmit information within neurons, trigger contractions within muscle cells, and lead to exocytosis in secretory cells. Key Concepts: All cells maintain a voltage difference across their plasma membranes. Action potentials are all-or-nothing, transient changes in membrane potentials of electrically excitable cells that carry important cellular information. Influx of sodium ions through voltage-gated sodium channels is responsible for the upstroke of the action potential, whereas efflux of potassium ions through voltage-gated potassium channels is responsible for the falling phase. Propagation of action potentials depends on gating kinetics of ion channels and intracellular and membrane resistances. Keywords: membrane potential; ionic current; threshold; refractoriness; length constant

Tactile reception in cyprinid fish (silver crucian carp Carassius auratus gibelio and common carp Cyprinus carpio) was investigated by noninvasive recording of action potentials from the head skin in response to rhythmic tactile stimulation. Responses shaped as electric potentials following synchronously with changes in the pressure force on the skin were recorded using chloro-silver electrodes. It was shown that fish, like all terrestrial vertebrates, have phasic and tonic tactile receptors. Three main types of tactile responses were distinguished: (1) tonic responses as potential fluctuations repeating the dynamics of the tactile stimulus, (2) phasic responses as on-, off-, and on-off potentials, and (3) slow deflections from the resting potential. Compound responses comprising simultaneously different types of responses were also recorded. The response latency averaging 6.1 ms for tonic and 4.4 ms for phasic responses was clearly independent of the stimulus direction (switch-on, switch-off). The threshold level of tactile sensitivity in the perioral and gular head areas was 0.05–0.2 g/mm2. Tactile receptors on the silver crucian carp and common carp heads exhibited zonal distribution patterns, with sensory areas differing in their tactile sensitivity. Most sensitive areas were on the upper and lower lips, in the perilabial skin and the gular area on the ventral side of the head. Less sensitive areas were situated rostrally and ventrally to the eyeball. The areas situated dorsally and caudally to the eyeball proved to be insensitive to the applied tactile stimulation. In all sensory areas on the fish head, most intense responses were evoked by sliding tactile stimuli. The distribution specificity of tactile sensory areas meets their functional significance for fish feeding behavior.

HCN channels regulate excitability and rhythmicity in the hippocampal CA1 pyramidal cells. Perturbation in the HCN channel current (Ih) is associated with neuropsychiatric disorders, such as autism spectrum disorders. Recently, protein arginine methyltransferase 7 (PRMT7) was shown to be highly expressed in the hippocampus, including the CA1 region. However, the physiological function of PRMT7 in the CA1 neurons and the relationship to psychiatric disorders are unclear. Here we showed that PRMT7 knockout (KO) mice exhibit hyperactivity and deficits in social interaction. The firing frequency of the CA1 neurons in the PRMT7 KO mice was significantly higher than that in the wild-type (WT) mice. Compared with the WT CA1 neurons, the PRMT7 KO CA1 neurons showed a more hyperpolarized resting potential and a higher input resistance, which were occluded by the Ih-current inhibitor ZD7288; these findings were consistent with the decreased Ih and suggested the contribution of Ih-channel dysfunction to the PRMT7 KO phenotypes. The HCN1 protein level was decreased in the CA1 region of the PRMT7 KO mice in conjunction with a decrease in the expression of Shank3, which encodes a core scaffolding protein for HCN channel proteins. A brief application of the PRMT7 inhibitor DS437 did not reproduce the phenotype of the PRMT7 KO neurons, further indicating that PRMT7 regulates Ih by controlling the channel number rather than the open probability. Moreover, shRNA-mediated PRMT7 suppression reduced both the mRNA and protein levels of SHANK3, implying that PRMT7 deficiency might be responsible for the decrease in the HCN protein levels by altering Shank3 expression. These findings reveal a key role for PRMT7 in the regulation of HCN channel density in the CA1 pyramidal cells that may be amenable to pharmacological intervention for neuropsychiatric disorders., Neuroscience: Finding the brakes in the autistic brain Disrupted expression of an ion channel that helps stabilize brain cell activity contributes to behavioral symptoms in mice resembling those seen in autism spectrum disorders (ASDs). Nerve cell firing depends on the right balance of ions inside and outside cells, and a channel protein called HCN helps establish ionic conditions that prevent excessive activity. Researchers led by Hana Cho and Ilmin Kwon of the Sungkyunkwan University School of Medicine, Suwon, South Korea have demonstrated that mice lacking another protein called PRMT7 exhibit reduced numbers of HCN channels in brain structures known to be affected in animal models of ASDs. These mice exhibit hyperactivity and social anxiety, presumably as a consequence of poor regulation of nerve cell firing. The authors propose that this PRMT7-HCN pathway may offer a fruitful target for the development of neuropsychiatric therapies.

Biological and mechanical systems, whether by their overuse or their aging, will inevitably fail. Hearing provides a poignant example of this with noise-induced and age-related hearing loss. Hearing loss is not unique to humans, however, and is experienced by all animals in the face of wild and eclectic differences in ear morphology and operation. Here we exploited the high throughput and accessible tympanal ear of the desert locust, Schistocerca gregaria (mixed sex) to rigorously quantify changes in the auditory system due to noise exposure (3 kHz pure tone at 126 dB SPL) and age. We analysed tympanal dispalcements, morphology of the auditory Müller’s organ and measured activity of the auditory nerve, the transduction current and electrophysiological properties of individual auditory receptors. We found that noise mildly and transiently changes tympanal displacements, decreases both the width of the auditory nerve and the transduction current recorded from individual auditory neurons. Whereas age – but not noise - decreases the number of auditory neurons and increases their resting potential. Multiple other properties of Müller’s organ were unaffected by either age or noise including: the number of supporting cells in Müller’s organ or the nerve, membrane resistance and capacitance of the auditory neurons. The sound-evoked activity of the auditory nerve decreased as a function of age and this decrease was exacerbated by noise, with the largest difference during the middle of their life span. This ‘middle-aged deafness’ pattern of hearing loss mirrors that found for humans exposed to noise early in their life.Key point summaryAge and noise lead to hearing loss.Tympanal displacements have a transient and delayed reduction after noise.The number of auditory neurons decreases with age. The width of the auditory nerve is reduced with noise exposure.Locusts repeatedly exposed to noise lost their hearing compared to silent controls, but their hearing became similar to old silent control locusts due to age-related hearing loss dominating.The electrophysiological properties of the auditory neurons and the transduction current remained unchanged for aged locusts but repeated noise exposure led to a cumulative decrease in the transduction current.

The recurrent attacks of weakness in hypokalemic periodic paralysis (HypoPP) are caused by failure to maintain the resting potential, with paradoxical depolarization in low K+. Remarkably, 24 out of 25 HypoPP mutations are R/X substitutions in S4 segments of voltage-sensing domains of CaV1.1 (70% of cases) or NaV1.4 (10% of cases). Expression studies in oocytes and murine muscle show anomalous gating pore leakage currents (ω-pore) for six of eight CaV1.1-HypoPP mutations, with one exception being the charge-conserving R897K. The proposed consensus pathomechanism, whereby a gating pore leak predisposes to paradoxical depolarization in low K+, is now verified by continuous recording of Vm. Selective measurement of voltage-dependent Ca2+ release, in “healthy appearing” HypoPP fibers, shows only a modest decrease in the Ca2+-dependent peak fluorescence (Oregon green 488/EGTA), and supports the notion that stabilizing Vrest will be sufficient to prevent low-K+–induced loss of force. In our knockin mouse models of HypoPP (CaV1.1-R528H and NaV1.4-R669H), pretreatment with K+-channel openers protects against the loss of force with a 2 mM K+ challenge. Alternatively, gene editing offers the possibility of sustained protection from attacks of weakness, and may prevent the late-onset permanent myopathy. In a proof-of-principle study of cultured myoblasts and in vivo electroporation, we show selective editing of the mutant HypoPP allele, without compromise of the WT allele, using CRISPR/Cas-mediated indel formation to destroy the HypoPP allele or a CRISPR/Cas base editor to correct the missense mutation.

OBJECTIVE: We sought to define how sensory neurotransmitters substance P and calcitonin gene-related peptide (CGRP) affect membrane potential of vascular smooth muscle and endothelium. METHODS: Microelectrodes recorded membrane potential of smooth muscle from pressurized mouse mesenteric arteries (diameter, ~150 μm) and in endothelial tubes. RESULTS: Resting potential was similar (~ −45 mV) for each cell layer. Substance P hyperpolarized smooth muscle and endothelium ~ −15 mV; smooth muscle hyperpolarization was abolished by endothelial disruption or NO synthase inhibition. Blocking K(Ca) channels (apamin + charybdotoxin) attenuated hyperpolarization in both cell types. CGRP hyperpolarized endothelium and smooth muscle ~ −30 mV; smooth muscle hyperpolarization was independent of endothelium. Blocking K(Ca) channels prevented hyperpolarization to CGRP in endothelium but not smooth muscle. Inhibiting K(ATP) channels with glibenclamide or genetic deletion of K(IR)6.1 attenuated hyperpolarization in smooth muscle but not endothelium. Pinacidil (K(ATP) channel agonist) hyperpolarized smooth muscle more than endothelium (~ −35 vs. ~ −20 mV). CONCLUSIONS: Calcitonin gene-related peptide elicits greater hyperpolarization than substance P. Substance P hyperpolarizes both cell layers through K(Ca) channels and involves endothelium-derived NO in smooth muscle. Endothelial hyperpolarization to CGRP requires K(Ca) channels, while K(ATP) channels mediate hyperpolarization in smooth muscle. Differential K(+) channel activation in smooth muscle and endothelium through sensory neurotransmission may selectively tune mesenteric blood flow.