Sodium- and Calcium-Dependent Excitability of Embryonic Leech Ganglion Cells in Culture

1991 ◽  
Vol 155 (1) ◽  
pp. 435-453
Author(s):  
KARIN SCHIRRMACHER ◽  
JOACHIM W. DEITMER
Keyword(s):  
Cultured Cells ◽  
Ganglion Cells ◽  
Na Current ◽  
Calcium Dependent ◽  
Na Currents ◽  
Voltage Dependent ◽  
Inward Currents ◽  
Cell Current ◽  
K Currents

Voltage-dependent Na+ and Ca2+ inward currents underlying the action potential in cultured embryonic ganglion cells of the leech Hirudo medicinalis have been investigated using the gigaseal whole-cell current or voltage-clamp technique. Dissociated ganglion cells were isolated from 7- to 14-day-old embryos, and maintained in primary culture for up to 5 days. More than 95% of the cultured cells had voltage-dependent K+ currents and about 75% of the cells had voltagedependent inward currents. Action potentials of 60mV amplitude and 4 ms duration, similar to those in embryonic nerve cells in vivo, could be recorded. Three types of inward currents occurred in these cells: (1) an initial Na+ current, which activated and inactivated rapidly; (2) a second Na+ current, which activated slowly and persisted during membrane depolarization, showing very little inactivation, and (3) a Ca2+-dependent inward current. Both types of Na+ currents were resistant to tetrodotoxin (TTX, 0.2-5 μmol l−1). The Ca+ current was also carried by Ba2+, and was blocked by cobalt and cadmium. The fast Na+ current was first expressed in cells from 8-day-old embryos, 1 day earlier than the Ca2+ current. Between days 8 and 14 the density of the fast Na+ current increased from 22±3 to 51±6 μA cm−2 (±S.D., N=11), while the Ca2+ current grew from 10 μA cm−1 (N=2) to 15±4 μA cm−2 (N=10) during this time.

scholarly journals Voltage-dependent and odorant-regulated currents in isolated olfactory receptor neurons of the channel catfish.

1992 ◽  
Vol 99 (4) ◽  
pp. 505-529 ◽  
Author(s):  
T Miyamoto ◽  
D Restrepo ◽  
J H Teeter
Keyword(s):  
Action Potentials ◽  
Olfactory Receptor Neurons ◽  
Olfactory Neurons ◽  
Outward Currents ◽  
Na Currents ◽  
Voltage Dependent ◽  
Inward Currents ◽  
K Current ◽  
Receptor Neurons ◽  
K Currents

The electrical properties of olfactory receptor neurons, enzymatically dissociated from the channel catfish (Ictalurus punctatus), were studied using the whole-cell patch-clamp technique. Six voltage-dependent ionic currents were isolated. Transient inward currents (0.1-1.7 nA) were observed in response to depolarizing voltage steps from a holding potential of -80 mV in all neurons examined. They activated between -70 and -50 mV and were blocked by addition of 1 microM tetrodotoxin (TTX) to the bath or by replacing Na+ in the bath with N-methyl-D-glucamine and were classified as Na+ currents. Sustained inward currents, observed in most neurons examined when Na+ inward currents were blocked with TTX and outward currents were blocked by replacing K+ in the pipette solution with Cs+ and by addition of 10 mM Ba2+ to the bath, activated between -40 and -30 mV, reached a peak at 0 mV, and were blocked by 5 microM nimodipine. These currents were classified as L-type Ca2+ currents. Large, slowly activating outward currents that were blocked by simultaneous replacement of K+ in the pipette with Cs+ and addition of Ba2+ to the bath were observed in all olfactory neurons examined. The outward K+ currents activated over approximately the same range as the Na+ currents (-60 to -50 mV), but the Na+ currents were larger at the normal resting potential of the neurons (-45 +/- 11 mV, mean +/- SD, n = 52). Four different types of K+ currents could be differentiated: a Ca(2+)-activated K+ current, a transient K+ current, a delayed rectifier K+ current, and an inward rectifier K+ current. Spontaneous action potentials of varying amplitude were sometimes observed in the cell-attached recording configuration. Action potentials were not observed in whole-cell recordings with normal internal solution (K+ = 100 mM) in the pipette, but frequently appeared when K+ was reduced to 85 mM. These observations suggest that the membrane potential and action potential amplitude of catfish olfactory neurons are significantly affected by the activity of single channels due to the high input resistance (6.6 +/- 5.2 G omega, n = 20) and low membrane capacitance (2.1 +/- 1.1 pF, n = 46) of the cells.(ABSTRACT TRUNCATED AT 400 WORDS)

scholarly journals Kinetic analysis of the action of Leiurus scorpion alpha-toxin on ionic currents in myelinated nerve.

1985 ◽  
Vol 86 (5) ◽  
pp. 739-762 ◽  
Author(s):  
G K Wang ◽  
G Strichartz
Keyword(s):  
Steady State ◽  
Ionic Currents ◽  
Na Channel ◽  
Dependent Manner ◽  
Na Current ◽  
Toxin Concentration ◽  
Toxin Molecule ◽  
Channel Inactivation ◽  
Na Currents ◽  
Voltage Dependent

The effects of a neurotoxin, purified from the venom of the scorpion Leiurus quinquestriatus, on the ionic currents of toad single myelinated fibers were studied under voltage-clamp conditions. Unlike previous investigations using crude scorpion venom, purified Leiurus toxin II alpha at high concentrations (200-400 nM) did not affect the K currents, nor did it reduce the peak Na current in the early stages of treatment. The activation of the Na channel was unaffected by the toxin, the activation time course remained unchanged, and the peak Na current vs. voltage relationship was not altered. In contrast, Na channel inactivation was considerably slowed and became incomplete. As a result, a steady state Na current was maintained during prolonged depolarizations of several seconds. These steady state Na currents had a different voltage dependence from peak Na currents and appeared to result from the opening of previously inactivated Na channels. The opening kinetics of the steady state current were exponential and had rates approximately 100-fold slower than the normal activation processes described for transitions from the resting state to the open state. In addition, the dependence of the peak Na current on the potential of preceding conditioning pulses was also dramatically altered by toxin treatment; this parameter reached a minimal value near a membrane potential of -50 mV and then increased continuously to a "plateau" value at potentials greater than +50 mV. The amplitude of this plateau was dependent on toxin concentration, reaching a maximum value equal to approximately 50% of the peak current; voltage-dependent reversal of the toxin's action limits the amplitude of the plateauing effect. The measured plateau effect was half-maximum at a toxin concentration of 12 nM, a value quite similar to the concentration producing half of the maximum slowing of Na channel inactivation. The results of Hill plots for these actions suggest that one toxin molecule binds to one Na channel. Thus, the binding of a single toxin molecule probably both produces the steady state currents and slows the Na channel inactivation. We propose that Leiurus toxin inhibits the conversion of the open state to inactivated states in a voltage-dependent manner, and thereby permits a fraction of the total Na permeability to remain at membrane potentials where inactivation is normally complete.

Distribution of Na+ and K+ Currents in Soma, Axons and Growth Cones of Leech Retzius Neurones in Culture

1990 ◽  
Vol 150 (1) ◽  
pp. 1-17 ◽  
Author(s):  
U. GARCÍA ◽  
S. GRUMBACHER-REINERT ◽  
R. BOOKMAN ◽  
H. REUTER
Keyword(s):  
Cultured Cells ◽  
Growth Cones ◽  
Proximal Segment ◽  
Mapping Procedure ◽  
Na Currents ◽  
K Current ◽  
Current Densities ◽  
A New Technique ◽  
Retzius Cells ◽  
K Currents

1. Leech Retzius neurones were isolated by a new technique which allowed investigation of macroscopic currents over the surface of the cell body and the axons using loose patch-clamp. The distribution of ion current densities was measured for neurones that had just been removed from the CNS, and for cultured cells in which neurite outgrowth had begun. To standardize the mapping procedure, the same patch electrode was used at various sites along the neurone. 2. Immediately after isolation of the cell, rapidly activating and inactivating Na+ currents were recorded from distal segments of the axons, but not from the soma or the proximal segment. Na+ currents were isolated by using patch electrodes containing tetraethylammonium (TEA+) and 4-aminopyridine (4-AP) to block K+ channels and Cd2+ to block calcium channels. Na+ currents in all regions of the neurone where they could be recorded were similar in their voltage dependence and kinetics. The Na+ current density was highest at the broken tips of the axon stumps. 3. Neurites began to extend from the broken axon tips approximately 30min after isolation. Newly grown processes showed a high density of Na+ currents at their growth cones. After 2 days in culture the current densities became more uniformly distributed and Na+ currents could then be recorded in the soma and proximal axon segments. 4. In agreement with earlier studies made with conventional two-electrode voltage-clamp, three principal K+ currents were detected in Retzius cells: a rapidly activating and inactivatingA-type current blocked by 4-AP (IA); a more slowly activating and inactivating delayed K+ current blocked by TEA+ (IK1); and a Ca2+-activated K+ current (IC). Immediately after isolation of the Retzius cell, both rapid A-type and slow delayed K+ currents were distributed more uniformly than Na+ currents over the soma and axons. In their voltage sensitivities and kinetics, these two K+ currents were markedly different from each other; their characteristics were, however, constant in different regions of the cell. 5. Ca2+ currents were too small to be measured directly during depolarizing pulses. However, tail currents were large enough to demonstrate the presence of Ca2+ channels in the proximal segment of the axon and in the soma; the currents were not sufficiently large to resolve their spatial distribution. 6. It is concluded that ion channels are present in newly grown membranes and that the density of Na+ channels is highest in the tips of distal axon stumps from which outgrowth begins. By contrast, K+ currents are distributed more uniformly along the neurone.

Somatostatin increases voltage-dependent potassium currents in rat somatotrophs

1990 ◽  
Vol 259 (6) ◽  
pp. C854-C861 ◽  
Author(s):  
C. Chen ◽  
J. Zhang ◽  
J. D. Vincent ◽  
J. M. Israel
Keyword(s):  
Growth Hormone ◽  
Cell Voltage ◽  
Inhibitory Effect ◽  
Delayed Rectifier ◽  
Hormone Release ◽  
Growth Hormone Release ◽  
Voltage Dependent ◽  
Inward Currents ◽  
K Current ◽  
K Currents

To study the modulatory effects of somatostatin on membrane K+ currents, whole cell voltage-clamp recordings were performed on identified rat somatotrophs in primary culture. In the presence of Co2+ (2 mM) and tetrodotoxin (1 microM) in the bath solution to block Ca2+ and Na+ inward currents, two types of voltage-activated K+ currents were identified on the basis of their kinetics and pharmacology. First, a delayed rectifier K+ current (IK) had a threshold of -20 mV, did not decay during voltage steps lasting 300 ms, and was markedly attenuated by extracellular application of tetraethylammonium (TEA, 10 mM). Second, a transient outward K+ current (IA) was activated at -40 mV (from a holding potential of -80 mV) and persisted despite the presence of TEA. This IA was blocked by 4-aminopyridine (2 mM). Somatostatin (10 nM) increased IK by 75% and IA by 45% without obvious effects on steady-state voltage dependency of activation or inactivation, and these effects were reversible. This increase in K+ currents may contribute in part to the inhibitory effect of somatostatin on growth hormone release.

Fast Amplification of Dynamic Synaptic Inputs in Spinal Motoneurons In Vivo

2006 ◽  
Vol 96 (5) ◽  
pp. 2200-2206 ◽  
Author(s):  
Sarah M. Jones ◽  
Robert H. Lee
Keyword(s):  
High Frequency ◽  
Tendon Vibration ◽  
Spinal Motoneurons ◽  
Synaptic Inputs ◽  
State Response ◽  
Voltage Dependent ◽  
Inward Currents ◽  
The Difference ◽  
Adult Cat

The ability of voltage-dependent inward currents (likely Na+) of the adult cat lumbar motoneurons to amplify rapidly changing (i.e., dynamic) synaptic inputs was investigated using in vivo intracellular recording techniques. Fast amplification was assessed by measuring the magnitude of the high-frequency (180 Hz) component of the Ia synaptic input due to tendon vibration as a function of somatic voltage and was compared with the previously observed amplification of steady inputs (steady state response of PICs to slow inputs). Data from 17 experiments show that amplification of the dynamic input indeed occurred and was directly linked to neuromodulatory drive (standard state: decerebrate with intact descending neuromodulatory systems vs. minimal state: pentobarbital with said systems significantly inhibited). Fast amplification factors averaged 2.0 ± 0.7 (mean ± SD) in the standard neuromodulatory state. That is, the effective synaptic current was nearly twice as large at its peak as it was at hyperpolarized levels, ranging as high as 2.6. Although fast amplification was often smaller than the amplification of steady inputs, the difference was not statistically significant. However, the voltage at which fast amplification began was ∼10 mV more depolarized ( P < 0.01). It is concluded that both dynamic and steady inputs can be amplified, but there may be differences in mechanism.

scholarly journals Membrane properties of isolated mudpuppy taste cells.

1988 ◽  
Vol 91 (3) ◽  
pp. 351-371 ◽  
Author(s):  
S C Kinnamon ◽  
S D Roper
Keyword(s):  
Bitter Taste ◽  
Taste Receptor ◽  
Membrane Properties ◽  
Basal Cells ◽  
Patch Clamp Technique ◽  
Inward Current ◽  
Taste Cells ◽  
Voltage Dependent ◽  
Inward Currents ◽  
K Currents

The voltage-dependent currents of isolated Necturus lingual cells were studied using the whole-cell configuration of the patch-clamp technique. Nongustatory surface epithelial cells had only passive membrane properties. Small, spherical cells resembling basal cells responded to depolarizing voltage steps with predominantly outward K+ currents. Taste receptor cells generated both outward and inward currents in response to depolarizing voltage steps. Outward K+ currents activated at approximately 0 mV and increased almost linearly with increasing depolarization. The K+ current did not inactivate and was partially Ca++ dependent. One inward current activated at -40 mV, reached a peak at -20 mV, and rapidly inactivated. This transient inward current was blocked by tetrodotoxin (TTX), which indicates that it is an Na+ current. The other inward current activated at 0 mV, peaked at 30 mV, and slowly inactivated. This more sustained inward current had the kinetic and pharmacological properties of a slow Ca++ current. In addition, most taste cells had inwardly rectifying K+ currents. Sour taste stimuli (weak acids) decreased outward K+ currents and slightly reduced inward currents; bitter taste stimuli (quinine) reduced inward currents to a greater extent than outward currents. It is concluded that sour and bitter taste stimuli produce depolarizing receptor potentials, at least in part, by reducing the voltage-dependent K+ conductance.

Studies of solitary semicircular canal hair cells in the adult pigeon. II. Voltage-dependent ionic conductances

1989 ◽  
Vol 62 (4) ◽  
pp. 935-945 ◽  
Author(s):  
D. G. Lang ◽  
M. J. Correia
Keyword(s):  
Hair Cells ◽  
Semicircular Canal ◽  
Potassium Conductance ◽  
Outward Current ◽  
Resting Membrane Potential ◽  
Calcium Dependent ◽  
Ionic Conductances ◽  
Voltage Dependent ◽  
Steady State Inactivation ◽  
Inward Currents

1. The ionic conductances present in putative type II hair cells enzymatically dissociated from the anterior, posterior, and lateral semicircular canal cristae of the white king pigeon (Columba livia) vestibule were studied under whole cell voltage clamp. 2. Two classes of voltage-dependent potassium conductances were distinguishable on the basis of the time course of activation and inactivation and pharmacologic sensitivity. The rapid potassium conductance, IA, as inhibited by 6 mM 4-aminopyridine (4-AP), whereas the slow potassium conductance, IK, was inhibited by 50 mM tetraethylammonium (TEA). These conductances were not affected by extracellular calcium removal. IA was quite similar to the rapidly-inactivating A-current of molluscan soma, whereas IK was more like the delayed rectifier of molluscan soma. 3. The steady-state inactivation of IA occurred over a potential range from -100 to -40 mV. The threshold for activation of IA occurred between -60 and -50 mV. The slope conductance of the I-V curve over a range of -50 to -20 mV was 13.7 nS when the conditioning pulse was -100 mV, and we estimate it to be approximately 1-2 nS from the resting membrane potential of -56 mV. 4. The steady-state inactivation of IK was approximately 60% at -40 mV and was completely removed at -80 mV. The threshold for activation of IK was between -50 and -40 mV. The slope conductance of the I-V curve over a range of -50 to -20 mV was 10.5 nS when the conditioning pulse was -80 mV, and we estimate it to be approximately 6-7 nS from the resting potential of -56 mV. 5. At -56 mV (the average resting membrane potential of putative type II semicircular canal hair cells), approximately 10-14% of IA channels and approximately 57-70% of IK channels were not inactivated: thus IA and IK can contribute to the outward current during small depolarizations from rest. 6. A small calcium-dependent outward current, IK(Ca), could be elicited during step depolarizations from a holding potential of -40 mV. This calcium-dependent current was active over the range of -20 to +40 mV. 7. Inward currents could not be detected when the cells were exposed to normal physiological solutions. However, when the outward currents were blocked with internal cesium and the external solution contained 20 mM barium, sustained inward currents with rapid activation kinetics could be detected. The threshold for activation of the inward current occurred at -40 mV, and the I-V relationship peaked at -10 mV.(ABSTRACT TRUNCATED AT 400 WORDS)

Actions of diazoxide on CA1 neurons in hippoeampal slices from rats

1995 ◽  
Vol 73 (5) ◽  
pp. 608-618 ◽  
Author(s):  
G. Erdemli ◽  
K. Krnjević
Keyword(s):  
High Voltage ◽  
Outward Current ◽  
Inward Rectification ◽  
Delayed Rectifier ◽  
Sprague Dawley ◽  
Na Current ◽  
Ca1 Neurons ◽  
Voltage Dependent ◽  
Inward Currents ◽  
Ca Currents

Membrane effects of diazoxide (DZX) were examined in CA1 pyramidal neurons, mainly by whole-cell recording in slices kept at 33 °C (from Sprague–Dawley rats). Bath applications of DZX (0.65 mM) did not significantly change the resting input conductance; but instantaneous inward rectification was reduced by 47 ± 14% (near –110 mV). There was a similar depression of a large, sustained voltage-dependent outward current (by 44 ± 11% near 0 mV). A nearly identical reduction of the outward current recorded in a Ca current suppressing medium (but not in 30 mM tetraethylammonium) indicated that the DZX-sensitive current includes the delayed rectifier. In Mn, low-Ca medium containing tetraethylammonium and carbachol, DZX potentiated (by 43 ± 12%) the D-type slowly decaying outward current seen after hyperpolarizing pulses at a holding potential of ≈ −50 mV. DZX abolished or depressed slow inward currents, such as the tetrodotoxin-sensitive persistent Na current, high voltage activated Ca currents (IC50 = 0.47 mM), and the Q current. In 6 of 13 cells recorded with electrodes containing either guanosine or adenosine diphosphate, DZX potentiated the voitage-dependent outward current, but input conductances were reduced. In conclusion, although there was little indication that it activates classical KATP channels in CA1 neurons, DZX strongly depresses several voltage-dependent, slowly inactivating outward and inward currents, which are important modulators of cell excitability.Key words: KATP channels, persistent Na current, high voltage activated Ca currents, delayed rectifier, D current, sulphonylureas, nucleotide diphosphates.

Nicotinic Receptors of Different Species Exhibit Differential Sensitivities to Nitromethylene and Organophosphate Insecticides

1996 ◽  
Vol 24 (3) ◽  
pp. 367-376
Author(s):  
Marga Oortgiesen ◽  
Regina G.D.M. van Kleef ◽  
Ruud Zwart ◽  
Ingeborg van den Beukel ◽  
Henk P.M. Vijverberg
Keyword(s):  
Nicotinic Acetylcholine Receptors ◽  
Mammalian Cells ◽  
Acetylcholine Receptors ◽  
Experimental Approach ◽  
Cultured Cells ◽  
Ganglion Cells ◽  
Acetyl Cholinesterase ◽  
Mouse Muscle ◽  
Inward Currents ◽  
Neuronal Type

The effects of nitromethylene heterocycle (NMH) and organophosphate (OP) insecticides were studied on nicotinic acetylcholine receptors (nAChR) in cultured cells of different species origin, in order to examine the selectivity of these compounds at the level of the target sites. In mouse muscle BC3H1, mouse NIE-115 and human SH-SY5Y neuroblastoma, and locust thoracic ganglion cells, the neurotransmitter, acetylcholine (ACh), induces a similar transient inward current. Dependent on the cell type, the six NMHs acted as agonists and/or antagonists on nAChR. Distinct agonistic effects of NMHs on nAChR are observed on insect neurons only. Further, NMHs potently block nicotinic responses in insects, while mammalian cells are only moderately affected. In all cases, the neuronal type nAChR was more sensitive to blocking than the endplate type nAChR in mammalian cells. Parathion and paraoxon at micromolar concentrations inhibit ACh-induced nicotinic inward currents. The insecticide, parathion, is a 50-fold more potent inhibitor than its acetyl-cholinesterase-inhibiting metabolite, paraoxon. Moderate differences in sensitivity to the blocking action of the OPs appear to exist among cells of different species. The results demonstrate that the experimental approach of fundamental electrophysiology and the use of cell lines are relevant tools for investigating the species specificity of neurotoxic compounds.

Sign in / Sign up

Export Citation Format

Share Document