scholarly journals Ionic Species Involved in the Electrical Activity of Single Adult Aminergic Neurones Isolated from the Sixth Abdominal Ganglion of the Cockroach Periplaneta Americana

1989 ◽  
Vol 144 (1) ◽  
pp. 535-549 ◽  
Author(s):  
BRUNO LAPIED ◽  
CLAIRE O. MALÉCOT ◽  
MARCEL PELHATE
Keyword(s):  
Periplaneta Americana ◽  
Potassium Conductance ◽  
Ionic Species ◽  
Resting Potential ◽  
Channel Blockers ◽  
Patch Clamp Technique ◽  
Mgcl2 Concentration ◽  
Dorsal Area ◽  
Ca2 Channels

Adult neurones were obtained by dissociation of the dorsal area of the sixth abdominal (A6) ganglion of the cockroach, and electrical properties were studied with the patch-clamp technique. The neurones showed spontaneous fast action potentials, similar to those recorded with microelectrodes in neurones in situ along the dorsal median line of the A6 ganglion. Synthetic saxitoxin (sSTX) at concentrations of 10 × 10−8 to 1.0×10−7mol l−1 suppressed the action potential (AP) and induced a dose-dependent hyperpolarization of the resting potential, suggesting that two types of sSTX-sensitive Na+ channels are present. The resting potential was dependent on the external concentration of both Na+ and K+, with a similar sensitivity to each, yielding a slope of about 43 mV per 10-fold change in concentration. The delayed outward rectification present under control conditions was reduced by tetraethylammonium chloride (TEA-Cl, 1.0×10−2mol l−1). TEA-Cl or Ca2+-free saline abolished the afterhyperpolarization and increased the overshoot and duration of triggered APs, indicating that a calcium-activated potassium conductance contributes to the falling phase of the AP. At 3.0×10−3mol l−1, the Ca2+ channel blockers MnCl2, CoCl2 and NiCl2 lengthened the AP. A blocker-dependent increase in the overshoot and threshold of the AP and reduction of the afterhyperpolarization were observed, probably reflecting the relative potencies of these ions in blocking Ca2+ channels and thus the Ca2+- activated K+ conductance. Increasing MgCl2 concentration by 3.0 × 10−3mol l−1 had no effect on the AP, indicating that the positive shift of the threshold is due to the blockade of Ca2+ channels present at this potential. The results suggest that these isolated neurones are dorsal unpaired median neurones previously studied in a number of insect species.

Ca2+ Currents in Central Insect Neurons: Electrophysiological and Pharmacological Properties

1997 ◽  
Vol 77 (1) ◽  
pp. 186-199 ◽  
Author(s):  
Dieter Wicher ◽  
Heinz Penzlin
Keyword(s):  
Periplaneta Americana ◽  
Low Voltage ◽  
Fast Time ◽  
Channel Blockers ◽  
Pharmacological Properties ◽  
Patch Clamp Technique ◽  
Whole Cell Patch Clamp ◽  
Blocking Effects ◽  
Dum Neurons ◽  
Maximal Peak

Wicher, Dieter, and Heinz Penzlin. Ca2+ currents in central insect neurons: electrophysiological and pharmacological properties. J. Neurophysiol. 77: 186–199, 1997. Ca2+ currents in dorsal unpaired median (DUM) neurons isolated from the fifth abdominal ganglion of the cockroach Periplaneta americana were investigated with the whole cell patch-clamp technique. On the basis of kinetic and pharmacological properties, two different Ca2+ currents were separated in these cells: mid/low-voltage-activated (M-LVA) currents and high-voltage-activated (HVA) currents. M-LVA currents had an activation threshold of −50 mV and reached maximal peak values at −10 mV. They were sensitive to depolarized holding potentials and decayed very rapidly. The decay was largely Ca2+ dependent. M-LVA currents were effectively blocked by Cd2+ median inhibiting concentration (IC50 = 9 μM), but they also had a remarkable sensitivity to Ni2+ (IC50 = 19 μM). M-LVA currents were insensitive to vertebrate LVA channel blockers like flunarizine and amiloride. The currents were, however, potently blocked by ω-conotoxin MVIIC (1 μM) and ω-agatoxin IVA (50 nM). The blocking effects of ω-toxins developed fast (time constant τ = 15 s) and were fully reversible after wash. HVA currents activated positive to −30 mV and showed maximal peak currents at +10 mV. They were resistant to depolarized holding potentials up to −50 mV and decayed in a less pronounced manner than M-LVA currents. HVA currents were potently blocked by Cd2+ (IC50 = 5 μM) but less affected by Ni2+ (IC50 = 40 μM). These currents were reduced by phenylalkylamines like verapamil (10 μM) and benzothiazepines like diltiazem (10 μM), but they were insensitive to dihydropyridines like nifedipine (10 μM) and BAY K 8644 (10 μM). Furthermore, HVA currents were sensitive to ω-conotoxin GVIA (1 μM). The toxin-induced reduction of currents appeared slowly (τ ∼ 120 s) and the recovery after wash was incomplete in most cases. The dihydropyridine insensitivity of the phenylalkylamine-sensitive HVA currents is a property the cockroach DUM cells share with other invertebrate neurons. Compared with Ca2+ currents in vertebrates, the DUM neuron currents differ considerably from the presently known types. Although there are some similarities concerning kinetics, the pharmacological profile of the cockroach Ca2+ currents especially is very different from profiles already described for vertebrate currents.

scholarly journals Calcium Conductance in An Identified Cholinergic Synaptic Terminal in the Central Nervous System of the Cockroach

1987 ◽  
Vol 129 (1) ◽  
pp. 347-364
Author(s):  
JOHATHAN M. BLAGBURN ◽  
DAVID B. SATTELLE
Keyword(s):  
Periplaneta Americana ◽  
Resting Potential ◽  
Channel Blockers ◽  
Sensory Neurone ◽  
Axonal Membrane ◽  
K Channels ◽  
Large Numbers ◽  
Long Duration ◽  
The Central Nervous System ◽  
Conductance Increase

Intracellular microelectrodes were used to study a cholinergic synapse between two identified neurones: the lateral filiform hair sensory neurone (LFHSN) and giant interneurone 3 (GI 3) in the terminal ganglion of the first-instar cockroach Periplaneta americana. The presynaptic cell, LFHSN, was impaled in a region of the axon which forms large numbers of output synapses. The sign and magnitude of the LFHSN spike afterpotential were shown to depend on [Ca2+]o. l μmoll−1 tetrodotoxin (TTX) abolished LFHSN spikes but the addition of 0.1 mmoll−1 4-aminopyridine (4-AP) enabled regenerative depolarizations to be evoked which were followed by large EPSPs in GI3. Addition of 20mmol−1 tetraethylammonium ions (TEA+) abolished the cholinergic EPSPs but resulted in long-duration LFHSN spikes. Intracellular injection of caesium ions (Cs+) into LFHSN enabled long-duration spikes to be evoked and had no effect on synaptic transmission. Long-duration LFHSN spikes were (1) increased in amplitude by increased [Ca2+]o; (2) accompanied by an increase in conductance; (3) not abolished by replacement of external Na+ with Tris+ or choline+; (4) blocked by 1 mmoll−1 Cd2+ and 10 mmoll−1 Co2+; (5) not supported by substitution of Mg2+ for Ca2+; and (6) supported by Ba2+ substitution. They are thus considered to be Ca2+ spikes. The Ca2+ spikes were blocked by organic Ca2+ channel blockers at 0.5-1 mmoll−1. The putative Ca2+ spike was followed by a hyperpolarizing afterpotential (HAP), the duration of which was proportional to the amplitude and duration of the Ca2+ spike. The HAP was (1) accompanied by a conductance increase; (2) reversed at potentials 30mV more negative than resting potential; (3) not supported by substituting Ba2+ for Ca2+; and (4) partially blocked by 150 mmoll−1 TEA+. The HAP is considered to result from an increase in Ca2+-dependent K+ conductance. It is concluded that, in addition to Na+ channels and delayed rectifying K+ channels, Ca2+ channels and Ca2+-dependent K+ channels are present in the axonal membrane of LFHSN, in a region which forms many output synapses.

Role of Ca2+ and Na+ on luteinizing hormone release from the calf pituitary

1988 ◽  
Vol 255 (4) ◽  
pp. E469-E474
Author(s):  
J. P. Kile ◽  
M. S. Amoss
Keyword(s):  
Luteinizing Hormone ◽  
Ionophore A23187 ◽  
Channel Blockers ◽  
Hormone Release ◽  
Primary Cells ◽  
Rat Pituitary ◽  
Voltage Dependent ◽  
Lh Release ◽  
Ca2 Channels

It has been proposed that gonadotropin-releasing hormone (GnRH) stimulates Ca2+ entry by activation of voltage-independent, receptor-mediated Ca2+ channels in the rat gonadotroph. Little work has been done on the role of calcium in GnRH-induced luteinizing hormone (LH) release in species other than the rat. Therefore, this study was done to compare the effects of agents that alter Ca2+ or Na+ entry on LH release from calf anterior pituitary primary cells in culture. GnRH (100 ng/ml), Ca2+ ionophore A23187 (2.5 microM), and the depolarizing agent ouabain (0.1-10 microM) all produced significant increases (P less than 0.05) in LH release; these effects were significantly reduced when the cells were preincubated with the organic Ca2+ channel blockers nifedipine (1-10 microM) and verapamil (1-10 microM) and with Co2+ (0.01-1 mM). The effect of ouabain was inhibited by tetrodotoxin (TTX; 1-10 nM) as well as by nifedipine at 0.1-10 microM. In contrast to its effect on rat pituitary LH release, TTX significantly inhibited GnRH-stimulated LH release at 1-100 nM. These results suggest that GnRH-induced LH release may employ Ca2+ as a second messenger in bovine gonadotrophs and support recent speculation that GnRH-induced Ca2+ mobilization may in part be voltage dependent.

scholarly journals Thapsigargin inhibits voltage-activated calcium channels in adrenal glomerulosa cells

1993 ◽  
Vol 296 (2) ◽  
pp. 309-312 ◽  
Author(s):  
M F Rossier ◽  
C P Python ◽  
M M Burnay ◽  
W Schlegel ◽  
M B Vallotton ◽  
...  
Keyword(s):  
Cell Types ◽  
Inhibitory Effect ◽  
Zona Glomerulosa ◽  
High Threshold ◽  
Patch Clamp Technique ◽  
Dual Effect ◽  
Blocking Voltage ◽  
Current Voltage ◽  
Glomerulosa Cells ◽  
Ca2 Channels

Thapsigargin, an inhibitor of the microsomal Ca2+ pumps, has been extensively used to study the intracellular Ca2+ pool participating in the generation of the agonist-induced Ca2+ signal in various cell types. A dual effect of this agent was observed in bovine adrenal zona glomerulosa cells. At nanomolar concentrations, thapsigargin stimulated a sustained Ca2+ influx, probably resulting from Ca(2+)-store depletion. In contrast, when added at micromolar concentrations, thapsigargin prevented the rise in cytosolic free Ca2+ concentration ([Ca2+]c) induced by K+. This inhibitory effect of thapsigargin on voltage-activated Ca2+ channels was confirmed by measuring Ba2+ currents by the patch-clamp technique. Both low-threshold (T-type) and high-threshold (L-type) Ca2+ channels were affected by micromolar concentrations of thapsigargin. Analysis of the current-voltage relationship for T-type channels revealed that thapsigargin did not modify the sensitivity of these channels to the voltage, but decreased the maximal current flowing through the channels. In conclusion, thapsigargin appears to exert a dual effect on adrenal glomerulosa cells. At lower concentrations, this agent induces a sustained Ca2+ entry, whereas at higher concentrations it decreases [Ca2+]c by blocking voltage-activated Ca2+ channels.

Inward rectification in rat nucleus accumbens neurons

1989 ◽  
Vol 62 (6) ◽  
pp. 1280-1286 ◽  
Author(s):  
N. Uchimura ◽  
E. Cherubini ◽  
R. A. North
Keyword(s):  
Steady State ◽  
Nucleus Accumbens ◽  
Time Course ◽  
Potassium Conductance ◽  
Resting Potential ◽  
Inward Rectification ◽  
Resting Membrane Potential ◽  
Potential Range ◽  
Inward Current ◽  
Rat Nucleus Accumbens

1. Intracellular recordings were made from neurons in slices cut from the rat nucleus accumbens septi. Membrane currents were measured with a single-electrode voltage-clamp amplifier in the potential range -50 to -140 mV. 2. In control conditions (2.5 mM potassium), the resting membrane potential of the neurons was -83.4 +/- 1.1 (SE) mV (n = 157). Steady state membrane conductance was voltage dependent, being 34.8 +/- 1.7 nS (n = 25) at -100 mV and 8.0 +/- 0.7 nS (n = 25) at -60 mV. 3. Barium (1 microM) markedly reduced the inward rectification and caused a small inward current (40.6 +/- 8.7 pA, n = 8) at the resting potential. These effects became larger with higher barium concentrations, and, in 100 microM barium, the current-voltage relation was straight. 4. The block of the inward current by barium (at -130 mV) occurred with an exponential time course; the time constant was approximately 1 s at 1 microM barium and less than 90 ms with 100 microM. Strontium had effects similar to those of barium, but 1000-fold higher concentrations were required. Cesium chloride (2 mM) and rubidium chloride (2 mM) also blocked the inward rectification; their action reached steady state within 50 ms. 5. It is concluded that the nucleus accumbens neurons have a potassium conductance with many features of a typical inward rectifier and that this contributes to the potassium conductance at the resting potential.

Some properties of acetylcholine receptors in human cultured myotubes

1985 ◽  
Vol 224 (1235) ◽  
pp. 183-196 ◽  
Keyword(s):  
Acetylcholine Receptors ◽  
Single Channel ◽  
Fold Increase ◽  
Resting Potential ◽  
Patch Clamp Technique ◽  
Membrane Hyperpolarization ◽  
Human Myotubes ◽  
Cultured Myotubes ◽  
Channel Open Time ◽  
Ach Receptors

The distribution and single channel properties of acetylcholine (ACh) receptors in human myotubes grown in tissue culture have been examined. Radioautography of myotubes labelled with [ 125 I]α-bungarotoxin showed that ACh receptors are distributed uniformly over the myotube surface at a density of 3.9 ± 0.5 receptors per square micrometre. Ac­cumulations of ACh receptors (hot spots) were found rarely. The conductance and kinetics of ACh-activated channels were investi­gated with the patch-clamp technique. Cell-attached membrane patches were used in all experiments. A single channel conductance in the range 40–45 pS was calculated. No sublevels of conductance (substates) of the activated channel were observed. The distribution of channel open-times varied with ACh concentration. With 100 nM ACh, the distribution was best fitted by the sum of two exponentials, whereas with 1 μM ACh a single exponential could be fitted. The mean channel open-time at the myotube resting potential (ca. — 70 mV, 22°C) was 8.2 ms. The distribution of channel closed-times was complex at all concentrations of ACh studied (100 nM to 10 μm). With desensitizing doses of ACh (10 μM), channel openings occurred in obvious bursts; each burst usually appeared as part of a ‘cluster’ of bursts. Both burst duration and mean interval between bursts increased with membrane hyperpolarization. Individual channel open-times and burst durations showed similar voltage dependence (e-fold increase per 80 mV hyperpolarization), whereas both the channel closed-times within a burst and the number of openings per burst were independent of membrane potential.

scholarly journals Gonadotropin-Releasing Hormone Inhibits Ether-à-Go-Go-Related Gene K+ Currents in Mouse Gonadotropes

Endocrinology ◽  
2010 ◽  
Vol 151 (3) ◽  
pp. 1079-1088 ◽  
Author(s):  
Wiebke Hirdes ◽  
Crenguta Dinu ◽  
Christiane K. Bauer ◽  
Ulrich Boehm ◽  
Jürgen R. Schwarz
Keyword(s):  
Resting Potential ◽  
Related Gene ◽  
Activation Curve ◽  
Signal Cascade ◽  
Primary Mouse ◽  
Voltage Dependent ◽  
K Current ◽  
Unknown Signal ◽  
Ca2 Channels ◽  
K Currents

Secretion of LH from gonadotropes is initiated by a GnRH-induced increase in intracellular Ca2+ concentration ([Ca2+]i). This increase in [Ca2+]i is the result of Ca2+ release from intracellular stores and Ca2+ influx through voltage-dependent Ca2+ channels. Here we describe an ether-à-go-go-related gene (erg) K+ current in primary mouse gonadotropes and its possible function in the control of Ca2+ influx. To detect gonadotropes, we used a knock-in mouse strain, in which GnRH receptor-expressing cells are fluorescently labeled. Erg K+ currents were recorded in 80–90% of gonadotropes. Blockage of erg currents by E-4031 depolarized the resting potential by 5–8 mV and led to an increase in [Ca2+]i, which was abolished by nifedipine. GnRH inhibited erg currents by a reduction of the maximal erg current and in some cells additionally by a shift of the activation curve to more positive potentials. In conclusion, the erg current contributes to the maintenance of the resting potential in gonadotropes, thereby securing a low [Ca2+]i by restricting Ca2+ influx. In addition, the erg channels are modulated by GnRH by an as-yet unknown signal cascade.

Hyperexcitability at sites of nerve injury depends on voltage-sensitive Na+ channels

1994 ◽  
Vol 72 (1) ◽  
pp. 349-359 ◽  
Author(s):  
O. Matzner ◽  
M. Devor
Keyword(s):  
Nerve Injury ◽  
Duty Cycle ◽  
Firing Frequency ◽  
Na Channel ◽  
Channel Blockers ◽  
Na Channels ◽  
Experimental Conditions ◽  
K Channels ◽  
Inward Currents ◽  
Ca2 Channels

1. We used the tested fiber method to record from single myelinated afferents axons ending in a chronic nerve injury site (neuroma) in the rat sciatic nerve or L4,5 dorsal root. Axons were chosen for study that fired spontaneously with a stable tonic or interrupted (bursty) autorhythmic firing pattern. 2. Agents that block voltage-sensitive Na+ channels [tetrodotoxin (TTX), lidocaine], voltage-sensitive Ca2+ channels (Cd2+, Co2+, Ni2+, verapamil, D600, nifedipine, and fluarizine), volt-age-sensitive K+ channels [tetraethylammonium (TEA), 4-aminopyridine (4-AP)], and Ca(2+)-activated K+ channels (gK+Ca2+;quinidine, apamine) were applied topically to the neuroma. Effects on baseline rhythmogenesis and on the duty cycle of bursting were documented. Spike pattern analysis was used to determine whether changes in firing frequency were associated with changes in impulse initiation (electrogenesis), or resulted from (partial) block of impulse propagation downstream from the site of electrogenesis. Effects of veratridine were also noted. 3. Na+ channel blockers consistently quenched neuroma firing, and they did so by suppressing the process of impulse initiation. Only rarely was propagation block the dominant process. In bursty fibers the duration of on-periods shortened as the duration of off-periods lengthened, without a significant change in the baseline interspike interval (ISI). Veratridine accelerated firing, also via the impulse generating process. 4. Ca2+ channel blockers had essentially no effect on baseline firing rate (i.e., ISI). 5. Ca2+ channel blockers, as well as blockers of gK+Ca2+, had substantial, but inconsistent effects on burst pattern. It is not clear whether this reflects variability in the experimental conditions, or heterogeneity among the fibers sampled. 6. Blockade of K+ channels failed to evoke rhythmogenesis in acutely cut axons as it does in chronically injured axons, even in the presence of veratridine. This is consistent with other evidence that ectopic neuroma firing depends on postinjury remodeling of membrane electrical properties. 7. The data indicate that, in chronically injured axons, the inward currents that underly electrogenicity, enable ectopic discharge, and, together with outward K+ currents, set the fundamental firing rhythm (ISI), operate primarily with the use of voltage-sensitive Na+ rather than Ca2+ channels. 8. The on-off duty cycle in bursty fibers was affected by Na+ channel ligands and also, although less so, and less consistently by, Ca2+ channel ligands. This indicates that both may play a role in the slow modulations of membrane potential that presumably underly interrupted autorhythmicity.

Regulation of calcium current by voltage and cytoplasmic calcium in canine gastric smooth muscle

1992 ◽  
Vol 262 (3) ◽  
pp. C691-C700 ◽  
Author(s):  
F. Vogalis ◽  
N. G. Publicover ◽  
K. M. Sanders
Keyword(s):  
Time Course ◽  
Fast Component ◽  
Patch Clamp Technique ◽  
Similar Time ◽  
Whole Cell Patch Clamp ◽  
Gastric Smooth Muscle ◽  
Double Exponential ◽  
Circular Layer ◽  
Double Exponential Function ◽  
Ca2 Channels

The regulation of Ca2+ current by intracellular Ca2+ was studied in isolated myocytes from the circular layer of canine gastric antrum. Ca2+ current was measured with the whole cell patch-clamp technique, and changes in cytoplasmic Ca2+ ([Ca2+]i) were simultaneously measured with indo-1 fluorescence. Ca2+ currents were activated by depolarization and inactivated despite maintained depolarization. Ca2+ current inactivation was fit with a double exponential function. Using Ba2+ or Na+ as charge carriers removed the fast component of inactivation, whereas enhanced intracellular buffering of Ca2+ did not remove the fast component. Ca2+ currents were associated with a rise in [Ca2+]i. The decrease in [Ca2+]i following repolarization was exponential, and during the relaxation of [Ca2+]i, Ca2+ current was inactivated. The inward current recovered with a similar time course as the decrease in [Ca2+]i, suggesting that [Ca2+]i regulates the basal availability of Ca2+ channels. These data support the hypothesis that, although [Ca2+]i may influence the resting level of inactivation, it is the "submembrane" compartment of [Ca2+]i that regulates the development of inactivation.

scholarly journals Voltage-gated transient currents in bovine adrenal fasciculata cells. I. T-type Ca2+ current.

1993 ◽  
Vol 102 (2) ◽  
pp. 217-237 ◽  
Author(s):  
B Mlinar ◽  
B A Biagi ◽  
J J Enyeart
Keyword(s):  
Time Constant ◽  
Low Voltage ◽  
Zona Fasciculata ◽  
Patch Clamp Technique ◽  
Time Constants ◽  
Voltage Gated ◽  
Wide Range ◽  
Boltzmann Function ◽  
Voltage Dependent ◽  
Ca2 Channels

The whole cell version of the patch clamp technique was used to identify and characterize voltage-gated Ca2+ channels in enzymatically dissociated bovine adrenal zona fasciculata (AZF) cells. The great majority of cells (84 of 86) expressed only low voltage-activated, rapidly inactivating Ca2+ current with properties of T-type Ca2+ current described in other cells. Voltage-dependent activation of this current was fit by a Boltzmann function raised to an integer power of 4 with a midpoint at -17 mV. Independent estimates of the single channel gating charge obtained from the activation curve and using the "limiting logarithmic potential sensitivity" were 8.1 and 6.8 elementary charges, respectively. Inactivation was a steep function of voltage with a v1/2 of -49.9 mV and a slope factor K of 3.73 mV. The expression of a single Ca2+ channel subtype by AZF cells allowed the voltage-dependent gating and kinetic properties of T current to be studied over a wide range of potentials. Analysis of the gating kinetics of this Ca2+ current indicate that T channel activation, inactivation, deactivation (closing), and reactivation (recovery from inactivation) each include voltage-independent transitions that become rate limiting at extreme voltages. Ca2+ current activated with voltage-dependent sigmoidal kinetics that were described by an m4 model. The activation time constant varied exponentially at test potentials between -30 and +10 mV, approaching a voltage-independent minimum of 1.6 ms. The inactivation time constant (tau i) also decreased exponentially to a minimum of 18.3 ms at potentials positive to 0 mV. T channel closing (deactivation) was faster at more negative voltages; the deactivation time constant (tau d) decreased from 8.14 +/- 0.7 to 0.48 +/- 0.1 ms at potentials between -40 and -150 mV. T channels inactivated by depolarization returned to the closed state along pathways that included two voltage-dependent time constants. tau rec-s ranged from 8.11 to 4.80 s when the recovery potential was varied from -50 to -90 mV, while tau rec-f decreased from 1.01 to 0.372 s. At potentials negative to -70 mV, both time constants approached minimum values. The low voltage-activated Ca2+ current in AZF cells was blocked by the T channel selective antagonist Ni2+ with an IC50 of 20 microM. At similar concentrations, Ni2+ also blocked cortisol secretion stimulated by adrenocorticotropic hormone. Our results indicate that bovine AZF cells are distinctive among secretory cells in expressing primarily or exclusively T-type Ca2+ channels.(ABSTRACT TRUNCATED AT 400 WORDS)

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