Altered Excitability of Intestinal Neurons in Primary Culture Caused by Acute Oxidative Stress

2003 ◽  
Vol 89 (6) ◽  
pp. 3039-3050 ◽  
Author(s):  
Fivos Vogalis ◽  
John R. Harvey
Keyword(s):  
Oxidative Stress ◽  
Primary Culture ◽  
Large Amplitude ◽  
Action Potentials ◽  
Outward Current ◽  
Internal Perfusion ◽  
Inward Current ◽  
Cell Recording ◽  
Ionic Mechanisms ◽  
Inwardly Rectifying

Neurons were isolated from the intestine of guinea pigs and grown in primary culture for ≤15 days. Using conventional whole cell recording techniques, we demonstrated that the majority of neurons express a prolonged poststimulus afterhyperpolarization (slow AHP). These neurons also had large-amplitude (∼100 mV), broad-duration (∼2 ms) action potentials and generated a hyperpolarization activated inward current ( Ih). Application of H2O2 (0.22–8.8 mM) hyperpolarized these neurons but not those lacking slow AHPs. The H2O2-induced hyperpolarization was followed by irreversible depolarization at higher concentrations (more than ∼1 mM) of H2O2 while it was maintained after washout of submillimolar H2O2. The ionic mechanisms underlying the hyperpolarization included the suppression of Ih and the activation of an inwardly rectifying outward current, which was blocked by glybenclamide (25–50 μM) and TEA (30 mM). In addition, H2O2 suppressed the slow AHP and its underlying current. Internal perfusion of catalase and glutathione opposed the H2O2-mediated decrease in IsAHP. Our results indicate that acute oxidative stress has neuron- and conductance-specific actions in intestinal neurons that may underlie pathophysiological conditions.

Dopamine modulation of two subthreshold currents produces phase shifts in activity of an identified motoneuron

1995 ◽  
Vol 74 (4) ◽  
pp. 1404-1420 ◽  
Author(s):  
R. M. Harris-Warrick ◽  
L. M. Coniglio ◽  
R. M. Levini ◽  
S. Gueron ◽  
J. Guckenheimer
Keyword(s):  
Action Potentials ◽  
Voltage Dependence ◽  
Motor Pattern ◽  
Outward Current ◽  
Untreated Cell ◽  
Slight Reduction ◽  
Inward Current ◽  
Postinhibitory Rebound ◽  
Spike Latency ◽  
Dopamine Modulation

1. The lateral pyloric (LP) neuron is a component of the 14-neuron pyloric central pattern generator in the stomatogastric ganglion of the spiny lobster, Panulirus interruptus. In the pyloric rhythm, this neuron fires rhythmic bursts of action potentials whose phasing depends on the pattern of synaptic inhibition from other network neurons and on the intrinsic postinhibitory rebound properties of the LP cell itself. Bath-applied dopamine excites the LP cell and causes its activity to be phase advanced in the pyloric motor pattern. At least part of this modulatory effect is due to dopaminergic modulation of the intrinsic rate of postinhibitory rebound in the LP cell. 2. The LP neuron was isolated from all detectable synaptic input. We measured the rate of recovery after 1-s hyperpolarizing current injections of varying amplitudes, quantifying the latency to the first spike following the hyperpolarizing prepulse and the interval between the first and second action potentials. Dopamine reduced both the first spike latency and the first interspike interval (ISI) in the isolated LP neuron. During the hyperpolarizating pre-steps, the LP cell showed a slow depolarizing sag voltage that was enhanced by dopamine. 3. We used voltage clamp to analyze dopamine modulation of subthreshold ionic currents whose activity is affected by hyperpolarizing prepulses. Dopamine modulated the transient potassium current IA by reducing its maximal conductance and shifting its voltage dependence for activation and inactivation to more depolarized voltages. This outward current is normally transiently activated after hyperpolarization of the LP cell, and delays the rate of postinhibitory rebound; by reducing IA, dopamine thus accelerates the rate of rebound of the LP neuron. 4. Dopamine also modulated the hyperpolarization-activated inward current Ih by shifting its voltage dependence for activation 20 mV in the depolarizing direction and accelerating its rate of activation. This enhanced inward current helps accelerate the rate of rebound in the LP cell after inhibition. 5. The relative roles of Ih and IA in determining the first spike latency and first ISI were explored using pharmacological blockers of Ih (Cs+) and IA [4-aminopyridine (4-AP)]. Blockade of Ih prolonged the first spike latency and first ISI, but only slightly reduced the net effect of dopamine. In the continued presence of Cs+, blockade of IA with 4-AP greatly shortened the first spike latency and first ISI. Under conditions where both Ih and IA were blocked, dopamine had no additional effect on the LP cell. 6. We used the dynamic clamp technique to further study the relative roles of IA and Ih modulation in dopamine's phase advance of the LP cell. We blocked the endogenous Ih with Cs+ and replaced it with a simulated current generated by a computer model of Ih. The neuron with simulated Ih gave curves relating the hyperpolarizing prepulse amplitude to first spike latency that were the same as in the untreated cell. Changing the computer parameters of the simulated Ih to those induced by dopamine without changing IA caused only a slight reduction in first spike latency, which was approximately 20% of the total reduction caused by dopamine in an untreated cell. Bath application of dopamine in the presence of Cs+ and simulated Ih (with control parameters) allowed us to determine the effect of altering IA but not Ih: this caused a significant reduction in first spike latency, but it was still only approximately 70% of the effect of dopamine in the untreated cell. Finally, in the continued presence of dopamine, changing the parameters of the simulated Ih to those observed with dopamine reduced the first spike latency to that seen with dopamine in the untreated cell. 7. We generated a mathematical model of the lobster LP neuron, based on the model of Buchholtz et al. for the crab LP neuron.

Leptin and CCK modulate complementary background conductances to depolarize cultured nodose neurons

2006 ◽  
Vol 290 (2) ◽  
pp. C427-C432 ◽  
Author(s):  
J. H. Peters ◽  
R. C. Ritter ◽  
S. M. Simasko
Keyword(s):  
Food Intake ◽  
Outward Current ◽  
Vagal Afferents ◽  
Afferent Neurons ◽  
Inward Current ◽  
Adult Rat ◽  
Nodose Ganglia ◽  
Sodium Dependent ◽  
Ionic Mechanisms ◽  
Vagal Afferent

We have previously reported that intraceliac infusion of leptin induces a reduction of meal size that depends on intact vagal afferents. This effect of leptin is enhanced in the presence of cholecystokinin (CCK). The mechanisms by which leptin and CCK activate vagal afferent neurons are not known. In the present study, we have begun to address this question by using patch-clamp electrophysiological techniques to examine the mechanisms by which leptin and CCK activate cultured vagal afferents from adult rat nodose ganglia. We found that leptin depolarized 41 (60%) of 68 neurons. The magnitude of membrane depolarization was dependent on leptin concentration and occurred in both capsaicin-sensitive and capsaicin-insensitive neurons. We also found that a majority (16 of 22; 73%) of nodose neurons activated by leptin were also sensitive to CCK. CCK-induced depolarization was primarily associated with the increase of an inward current (11 of 12), whereas leptin induced multiple changes in background conductances through a decrease in an outward current (7 of 13), an increase in an inward current (3 of 13), or both (3 of 13). However, further isolation of background currents by recording in solutions that contained only sodium or only potassium revealed that both leptin and CCK were capable of increasing a sodium-dependent conductance or inhibiting a potassium-dependent conductance. Our results support the hypothesis that vagal afferents are a point of convergence and integration of leptin and CCK signaling for control of food intake and suggest multiple ionic mechanisms by which leptin and CCK activate vagal afferent neurons.

Membrane potential oscillations in molluscan “burster” neurones

1979 ◽  
Vol 81 (1) ◽  
pp. 93-112
Author(s):  
R. W. Meech
Keyword(s):  
Membrane Potential ◽  
Action Potential ◽  
Intracellular Ph ◽  
Action Potentials ◽  
Outward Current ◽  
Ionized Calcium ◽  
Isolated Cells ◽  
Potential Oscillations ◽  
Inward Current ◽  
Membrane Potential Oscillations

Membrane potential oscillations can be induced in molluscan neurones under a variety of artificial conditions. In the so-called ‘burster’ neurones oscillations are generated even in isolated cells. A likely mechanism for ‘bursting’ involves the following ionic currents: 1. A transient inward current carried by Na+ and Ca2+. This current is responsible for the upstroke of the action potentials. 2. A delayed outward current carried by K+. This current is voltage-sensitive and is responsible for the downstroke of the action potential during the early part of the burst. It becomes progressively inactivated during the burst. Its amplitude depends on the intracellular pH. 3. A rapidly developing outward current carried by K+ which is inactivated at potentials close to action potential threshold. This current tends to hold the membrane in the hyperpolarized state and is involved in spacing the action potentials. 4. A prolonged inward current which may not inactivate. It is probably carried by both Na+ and Ca2+. This current is responsible for the depolarizing phase of the burst but also contributes to the action potential. 5. A slowly developing outward current, carried by K+. This current appears as a result of a slow increase in intracellular ionized calcium and is responsible for the hyperpolarizing phase of the burst. Note that a transient increase in this current may also contribute to the falling phase of the action potential during the later stages of the burst. It is also sensitive to intracellular pH. One of the more significant features of this system of producing membrane potential oscillations is that the frequency of the bursts depends on the rate at which the intracellular ionized calcium returns to its resting level. This process depends on the metabolic state of the animal which can thereby exert a considerable influence on the electrical activity of burster neurones.

Role of Na-Ca exchange in the action potential changes caused by drive in cardiac myocytes exposed to different Ca2+ loads

1999 ◽  
Vol 77 (6) ◽  
pp. 383-397
Author(s):  
Qi-Ying Liu ◽  
Mario Vassalle
Keyword(s):  
Action Potentials ◽  
Ventricular Myocytes ◽  
Outward Current ◽  
Time Dependent ◽  
Inward Current ◽  
Tail Current ◽  
Outward Currents ◽  
Inward Currents ◽  
Single Ventricular Myocytes

The role of Na-Ca exchange in the membrane potential changes caused by repetitive activity ("drive") was studied in guinea pig single ventricular myocytes exposed to different [Ca2+]o. The following results were obtained. (i) In 5.4 mM [Ca2+]o, the action potentials (APs) gradually shortened during drive, and the outward current during a train of depolarizing voltage clamp steps gradually increased. (ii) The APs shortened more and were followed by a decaying voltage tail during drive in the presence of 5 mM caffeine; the outward current became larger and there was an inward tail current on repolarization during a train of depolarizing steps. (iii) These effects outlasted drive so that immediately after a train of APs, currents were already bigger and, after a train of steps, APs were already shorter. (iv) In 0.54 mM [Ca2+]o, the above effects were much smaller. (v) In high [Ca2+]o APs were shorter and outward currents larger than in low [Ca2+]o. (vi) In 10.8 mM [Ca2+]o, both outward and inward currents during long steps were exaggerated by prior drive, even with steps (+80 and +120 mV) at which there was no apparent inward current identifiable as ICa. (vii) In 0.54 mM [Ca2+]o, the time-dependent outward current was small and prior drive slightly increased it. (viii) During long steps, caffeine markedly increased outward and inward tail currents, and these effects were greatly decreased by low [Ca2+]o. (ix) After drive in the presence of caffeine, Ni2+ decreased the outward and inward tail currents. It is concluded that in the presence of high [Ca2+]o drive activates outward and inward Na-Ca exchange currents. During drive, the outward current participates in the plateau shortening and the inward tail current in the voltage tail after the action potential.Key words: ventricular myocytes, repetitive activity, outward and inward Na-Ca exchange currents, caffeine, nickel.

scholarly journals Loss of the K+ channel Kv2.1 greatly reduces outward dark current and causes ionic dysregulation and degeneration in rod photoreceptors

2021 ◽  
Vol 153 (2) ◽  
Author(s):  
Christopher Fortenbach ◽  
Gabriel Peinado Allina ◽  
Camilla M. Shores ◽  
Sarah J. Karlen ◽  
Eric B. Miller ◽  
...  
Keyword(s):  
Dark Current ◽  
Outward Current ◽  
Physiological Model ◽  
Resting Potential ◽  
K Channel ◽  
Inward Current ◽  
Signal Light ◽  
Ionic Mechanisms ◽  
Knockout Animals ◽  
Dark Currents

Vertebrate retinal photoreceptors signal light by suppressing a circulating “dark current” that maintains their relative depolarization in the dark. This dark current is composed of an inward current through CNG channels and NCKX transporters in the outer segment that is balanced by outward current exiting principally from the inner segment. It has been hypothesized that Kv2.1 channels carry a predominant fraction of the outward current in rods. We examined this hypothesis by comparing whole cell, suction electrode, and electroretinographic recordings from Kv2.1 knockout (Kv2.1−/−) and wild-type (WT) mouse rods. Single cell recordings revealed flash responses with unusual kinetics, and reduced dark currents that were quantitatively consistent with the measured depolarization of the membrane resting potential in the dark. A two-compartment (outer and inner segment) physiological model based on known ionic mechanisms revealed that the abnormal Kv2.1−/− rod photoresponses arise principally from the voltage dependencies of the known conductances and the NCKX exchanger, and a highly elevated fraction of inward current carried by Ca2+ through CNG channels due to the aberrant depolarization. Kv2.1−/− rods had shorter outer segments than WT and dysmorphic mitochondria in their inner segments. Optical coherence tomography of knockout animals demonstrated a slow photoreceptor degeneration over a period of 6 mo. Overall, these findings reveal that Kv2.1 channels carry 70–80% of the non-NKX outward dark current of the mouse rod, and that the depolarization caused by the loss of Kv2.1 results in elevated Ca2+ influx through CNG channels and elevated free intracellular Ca2+, leading to progressive degeneration.

Expression and functional properties of TRPM2 channels in dopaminergic neurons of the substantia nigra of the rat

2011 ◽  
Vol 106 (6) ◽  
pp. 2865-2875 ◽  
Author(s):  
Kenny K. H. Chung ◽  
Peter S. Freestone ◽  
Janusz Lipski
Keyword(s):  
Oxidative Stress ◽  
Substantia Nigra ◽  
Functional Properties ◽  
Transient Receptor Potential ◽  
Brain Slices ◽  
Disease Process ◽  
Outward Current ◽  
Substantia Nigra Pars Compacta ◽  
Inward Current ◽  
Trpm2 Channels

Transient receptor potential melastatin 2 (TRPM2) channels are sensitive to oxidative stress, and their activation can lead to cell death. Although these channels have been extensively studied in expression systems, their role in the brain, particularly in the substantia nigra pars compacta (SNc), remains unknown. In this study, we assessed the expression and functional properties of TRPM2 channels in rat dopaminergic SNc neurons, using acute brain slices. RT-PCR analysis revealed TRPM2 mRNA expression in the SNc region. Immunohistochemistry demonstrated expression of TRPM2 protein in tyrosine hydroxylase-positive neurons. Channel function was tested with whole cell patch-clamp recordings and calcium (fura-2) imaging. Intracellular application of ADP-ribose (50–400 μM) evoked a dose-dependent, desensitizing inward current and intracellular free calcium concentration ([Ca2+]i) rise. These responses were strongly inhibited by the nonselective TRPM2 channel blockers clotrimazole and flufenamic acid. Exogenous application of H2O2 (1–5 mM) evoked a rise in [Ca2+]i and an outward current mainly due to activation of ATP-sensitive potassium (KATP) channels. Inhibition of K+ conductance with Cs+ and tetraethylammonium unmasked an inward current. The inward current and/or [Ca2+]i rise were partially blocked by clotrimazole and N-( p-amylcinnamoyl)anthranilic acid (ACA). The H2O2-induced [Ca2+]i rise was abolished in “zero” extracellular Ca2+ concentration and was enhanced at higher baseline [Ca2+]i, consistent with activation of TRPM2 channels in the cell membrane. These results provide evidence for the functional expression of TRPM2 channels in dopaminergic SNc neurons. Given the involvement of oxidative stress in degeneration of SNc neurons in Parkinson's disease, further studies are needed to determine the pathophysiological role of these channels in the disease process.

Currents under voltage clamp of burst-forming neurons of the cardiac ganglion of the lobster (Homarus americanus)

1986 ◽  
Vol 56 (6) ◽  
pp. 1739-1762 ◽  
Author(s):  
K. Tazaki ◽  
I. M. Cooke
Keyword(s):  
Voltage Clamp ◽  
Action Potentials ◽  
Reversal Potential ◽  
Outward Current ◽  
Homarus Americanus ◽  
Resting Potential ◽  
Molluscan Neurons ◽  
Cardiac Ganglion ◽  
Inward Current ◽  
Tail Current

Crustacean cardiac ganglion neuronal somata, although incapable of generating action potentials, produce regenerative, slow (greater than 200 ms) depolarizing potentials reaching -20 mV (from -50 mV) in response to depolarizing stimuli. These potentials initiate a burst of action potentials in the axon and are thus termed driver potentials. The somata of the anterior-most neurons (cells 1 or 2) were isolated by ligaturing for study of their membrane currents with a two-electrode voltage clamp. Inward current is attributed to Ca2+ by reason of dependence of driver potential amplitude on [Ca2+]0, independence of [Na+]0, resistance to tetrodotoxin, and inhibition by Cd (0.2 mM) and Mn (4 mM). Ca-mediated current (ICa) is present at -40 mV. It is optimally activated by a holding potential (Vh) of -50 to -60 mV and by clamps (command potential, Vc) to -10 mV. Time to peak (10-30 ms) and amplitude are strongly voltage dependent. Maximum tail-current amplitudes observed at -70 to -85 mV are ca. 100 nA. Inward tail peaks may not be resolved by our clamp (settling time, 2 ms). Tails relax with a time constant (tau) of approximately equal to 12 ms (at -70 to -85 mV). ICa exhibits inactivation in double pulse regimes. Recovery has a tau of approximately equal to 0.7 s. Tail current analyses indicate an exponential decline (tau approximately equal to 23 ms at -20 mV) toward a maintained amplitude of inward current tails. Analysis of outward currents indicates the presence of three conductance mechanisms having voltage dependences, time courses, and pharmacology similar to those of early outward current (IA), delayed outward current (IK), and outward current (IC) of molluscan neurons. Analysis of tail currents indicates a reversal potential for each of these near -75 mV, indicating that they are K currents. Early outward current, IA, shows a peak at 5 ms followed by rapid decline. Response to a second clamp given within 0.4 s is reduced; recovery is exponential, with a tau of approximately equal to 200 ms (at Vh = -50 mV). The amplitude of IA tested at 0 mV shows activation or deactivation by subthreshold shifts of Vh. The extent and rate of these changes shows voltage dependence (tau approximately equal to 100-500 ms for subthreshold prepulses). At the normal cell resting potential of -50 mV the amplitude of IA is 25% of that tested from -80 mV.(ABSTRACT TRUNCATED AT 400 WORDS)

Roles of Ca2+ and Na+ in the inward current and action potentials of guinea pig ureteral myocytes

1997 ◽  
Vol 272 (2) ◽  
pp. C535-C542 ◽  
Author(s):  
J. L. Sui ◽  
C. Y. Kao
Keyword(s):  
Guinea Pig ◽  
Action Potentials ◽  
Outward Current ◽  
Membrane Conductance ◽  
Membrane Current ◽  
Resting Potential ◽  
Inward Current ◽  
K Current ◽  
Free Media ◽  
Ca2 Channels

Physiological roles of Ca2+ vs. Na+ in membrane currents and action potentials of ureteral myocytes were investigated on freshly dissociated guinea pig ureteral myocytes with the patch-clamp method. The myocytes are spindle shaped, with cell volume of 2,473 microm3, surface area of 2,014 microm2, capacitance of 48.2 pF, resting potential of -47.9 mV, and membrane conductance of 840 pS. The membrane current consists of a slow inward Ca2+ current (ICa) conducted by L-type Ca2+ channels and an actively fluctuating Ca2+-activated K+ current [IK(Ca)] conducted by Ca2+-activated maxi-K+ channels. ICa dominates the membrane current by being long lasting and more active at less depolarized potentials than IK(Ca) and by regulating IK(Ca). Ca2+-free media, Co2+, and nifedipine reduce or block ICa, whereas high extracellular Ca2+ concentration and BAY K 8644 enhance it. Action potential amplitudes and plateaus are regulated correspondingly. Related changes are also seen in IK(Ca) In contrast, no fast inward current attributable to Na+ was found. Replacing extracellular Na+ with tris(hydroxymethyl)aminomethane had no apparent effects on the inward or outward current or on the action potentials.

scholarly journals Multiple effects of calcium antagonists on plateau currents in cardiac Purkinje fibers.

1975 ◽  
Vol 66 (2) ◽  
pp. 169-192 ◽  
Author(s):  
R S Kass ◽  
R W Tsien
Keyword(s):  
Action Potentials ◽  
Membrane Surface ◽  
Outward Current ◽  
Inhibitory Influence ◽  
Inward Current ◽  
Purkinje Fibers ◽  
Cardiac Purkinje Fibers ◽  
Ca Antagonist ◽  
Ca Ions ◽  
Membrane Surface Charge

We studied the influence of Mn, La, and D600 on action potentials and plateau currents in cardiac Purkinje fibers. The Ca antagonists each abolished the second inward current, but they failed to act selectively. Voltage clamp experiments revealed two additional effects: decrease of slow outward current (iotachi) activation, and increase of net outward time-independent plateau current. These effects occurred at inhibitor concentrations used in earlier studies, and were essential to the reconstruction of observed Ca antagonist effects on electrical activity. The inhibitory influence of Mn, La, and D600 on iotachi suggested that iotachi activation might depend upon prior Ca entry. This hypothesis was not supported, however, when [Ca]omicron was varied: elevating [Ca]omicron enhanced Ca entry, but iotachi was nevertheless depressed. Thus, the results suggested instead that Ca antagonists and Ca ions have rather similar effects on iotachi, possibly mediated by changes in membrane surface charge.

G alpha o1 decapeptide modulates the hippocampal 5-HT1A potassium current

1995 ◽  
Vol 74 (5) ◽  
pp. 2189-2193 ◽  
Author(s):  
S. Oleskevich
Keyword(s):  
G Protein ◽  
Potassium Current ◽  
Granule Cells ◽  
Outward Current ◽  
Intracellular Perfusion ◽  
Electrophysiological Response ◽  
C Terminus ◽  
Bath Application ◽  
Cell Recording ◽  
Inwardly Rectifying

1. The serotonin1A (5-HT1A) receptor is coupled to an inwardly rectifying potassium current (IKir) via a G protein. The identity of the G-protein subtype was investigated with 2 10-amino acid peptides derived from the carboxyl (C) terminus of the alpha-subunits of the Go1 and Gi2 proteins (G alpha o1 and G alpha i2). The synthetic decapeptides were applied by intracellular perfusion during whole cell recording from dentate granule cells in the hippocampal slice preparation. 2. Bath application of 5-HT produced an IKir, which was blocked by the selective 5-HT1A receptor antagonist, pindobind5-HT1A. The G alpha o1 peptide inhibited the 5-HT1A IKir by 60 +/- 7% (mean +/- SE; t = 30 min), whereas the G alpha i2 peptide had no effect. The G alpha o1 peptide produced a slowly developing outward current that was not observed in the absence of peptide or in the presence of the G alpha i2 peptide. 3. The results indicate that G alpha o1 and not G alpha i2 modulates the 5-HT1A IKir in hippocampal granule cells. They also suggest that G alpha o1 occludes the 5-HT1A response by direct activation of the IKir. The intracellular perfusion of synthetic G alpha peptides provides a new approach to identify the G-protein subtype(s) in a receptor-mediated electrophysiological response.

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