Low-Voltage Activated T-Type Calcium Currents Are Differently Expressed in Superficial and Deep Layers of Guinea PigPiriform Cortex

1998 ◽  
Vol 79 (2) ◽  
pp. 808-816 ◽  
Author(s):  
Jacopo Magistretti ◽  
Marco de Curtis
Keyword(s):  
Guinea Pig ◽  
Calcium Current ◽  
Low Voltage ◽  
Piriform Cortex ◽  
Neuronal Firing ◽  
Calcium Currents ◽  
Patch Clamp Technique ◽  
Voltage Range ◽  
Deep Layers ◽  
Group 2

Magistretti, Jacopo and Marco de Curtis. Low-voltage activated T-type calcium currents are differently expressed in superficial and deep layers of guinea pig piriform cortex. J. Neurophysiol. 79: 808–816, 1998. A variety of voltage-dependent calcium conductances are known to control neuronal excitability by boosting peripheral synaptic potentials and by shaping neuronal firing patterns. The existence and functional significance of a differential expression of low- and high-voltage activated (LVA and HVA, respectively) calcium currents in subpopulations of neurons, acutely isolated from different layers of the guinea pig piriform cortex, were investigated with the whole cell variant of the patch-clamp technique. Calcium currents were recorded from pyramidal and multipolar neurons dissociated from layers II, III, and IV. Average membrane capacitance was larger in layer IV cells [13.1 ± 6.2 (SD) pF] than in neurons from layers II and III (8.6 ± 2.8 and 7.9 ± 3.1 pF, respectively). Neurons from all layers showed HVA calcium currents with an activation voltage range positive to −40 mV. Neurons dissociated from layers III and IV showed an LVA calcium current with the biophysical properties of a T-type conductance. Such a current displayed the following characteristics: 1) showed maximal amplitude of 11–16 pA/pF at −30 mV, 2) inactivated rapidly with a time constant of ∼22 ms at −30 mV, and 3) was completely steady-state inactivated at −60 mV. Only a subpopulation of layer II neurons (group 2 cells; circa 18%) displayed an LVA calcium current similar to that observed in deep layers. The general properties of layer II-group 2 cells were otherwise identical to those of group 1 neurons. The present study demonstrates that LVA calcium currents are differentially expressed in neurons acutely dissociated from distinct layers of the guinea pig piriform cortex.

Characterization of voltage-dependent calcium currents in mouse motoneurons

1992 ◽  
Vol 68 (1) ◽  
pp. 85-92 ◽  
Author(s):  
M. Mynlieff ◽  
K. G. Beam
Keyword(s):  
Voltage Dependence ◽  
Low Voltage ◽  
Calcium Currents ◽  
Patch Clamp Technique ◽  
Maximal Amplitude ◽  
Time To Peak ◽  
Whole Cell Patch Clamp ◽  
Voltage Dependent ◽  
Channel Currents

1. Calcium channel currents were measured with the whole-cell patch clamp technique in cultured, identified mouse motoneurons. Three components of current were operationally defined on the basis of voltage dependence, kinetics, and pharmacology. 2. Test potentials to -50 mV or greater (10 mM external Ca2+) elicited a low-voltage activated T-type current that was transient (decaying to baseline in less than 200 ms) and had a relatively slow time to peak (20-50 ms). A 1-s prepulse to -45 mV produced approximately half-maximal inactivation of this T current. 3. Two high-voltage activated (HVA) components of current (1 transient and 1 sustained) were activated by test potentials to -20 mV or greater (10 mM external Ca2+). A 1-s prepulse to -35 mV produced approximately half-maximal inactivation of the transient component without affecting the sustained component. 4. When Ba2+ was substituted for Ca2+ as the charge carrier, activation of the HVA components was shifted in the hyperpolarizing direction, and the relative amplitude of the transient HVA component was reduced. 5. Amiloride (1-2 mM) caused a reversible, partial block of the T current without affecting the HVA components. 6. The dihydropyridine agonist isopropyl 4-(2,1,3-benzoxadiazol-4-yl)-1,4-dihydro-2,6-dimethyl-5-nitro-3- pyridine-carboxylate [(+)-SDZ 202-791, 100 nM-1 microM)] shifted the activation of the sustained component of HVA current to more negative potentials and increased its maximal amplitude. Additionally, (+)-SDZ 202-791 caused the appearance of a slowed component of tail current.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Effect of FMRFamide on voltage-dependent currents in identified centrifugal neurons of the optic lobe of the cuttlefish, Sepia officinalis

2020 ◽  
Author(s):  
Abdesslam Chrachri
Keyword(s):  
Visual Processing ◽  
Calcium Current ◽  
Optic Lobe ◽  
Low Voltage ◽  
Sodium Current ◽  
Calcium Currents ◽  
Delayed Rectifier ◽  
Outward Currents ◽  
Voltage Dependent ◽  
Inward Currents

AbstractWhole-cell patch-clamp recordings from identified centrifugal neurons of the optic lobe in a slice preparation allowed the characterization of five voltage-dependent currents; two outward and three inward currents. The outward currents were; the 4-aminopyridine-sensitive transient potassium or A-current (IA), the TEA-sensitive sustained current or delayed rectifier (IK). The inward currents were; the tetrodotoxin-sensitive transient current or sodium current (INa). The second is the cobalt- and cadmium-sensitive sustained current which is enhanced by barium and blocked by the dihydropyridine antagonist, nifedipine suggesting that it could be the L-type calcium current (ICaL). Finally, another transient inward current, also carried by calcium, but unlike the L-type, this current is activated at more negative potentials and resembles the low-voltage-activated or T-type calcium current (ICaT) of other preparations.Application of the neuropeptide FMRFamide caused a significant attenuation to the peak amplitude of both sodium and sustained calcium currents without any apparent effect on the transient calcium current. Furthermore, FMRFamide also caused a reduction of both outward currents in these centrifugal neurons. The fact that FMRFamide reduced the magnitude of four of five characterized currents could suggest that this neuropeptide may act as a strong inhibitory agent on these neurons.SummaryFMRFamide modulate the ionic currents in identified centrifugal neurons in the optic lobe of cuttlefish: thus, FMRFamide could play a key role in visual processing of these animals.

Depression of delayed outward K+ current by Co2+ in guinea pig ventricular myocytes

1991 ◽  
Vol 261 (1) ◽  
pp. C23-C31 ◽  
Author(s):  
Z. Fan ◽  
M. Hiraoka
Keyword(s):  
Guinea Pig ◽  
Ventricular Myocytes ◽  
Hill Coefficient ◽  
Maximal Response ◽  
Fluctuation Analysis ◽  
Dependent Manner ◽  
Surface Charges ◽  
Patch Clamp Technique ◽  
Voltage Range ◽  
K Current

Effects of Co2+ on the delayed outward K+ current (IK) in guinea pig ventricular myocytes were studied using the whole cell patch-clamp technique. IK was activated by depolarizing voltage pulses positive to -30 mV and reached half-maximal activation at +24 mV. Co2+ shifted the activation curve to a more depolarized voltage range in a concentration-dependent manner, with a Co2+ concentration at which half-maximal response occurs (IC50) of 8 mM and a saturation value of +38 mV. The voltage dependency of IK gatings showed a shift similar to that of activation. In both cases the shift could be explained by screening of surface potential. The density of total negative surface charges sensed by Co2+ was estimated to be 1 e/225 A2. Co2+ also reduced the fully activated IK [IK(full)], and the dose-response curve had a Hill coefficient of 0.5 and an IC50 of 1 mM at 0 mV. Depression of IK(full) was mainly voltage independent. The single-channel unitary current estimated by fluctuation analysis was approximately 0.1 pA at -30 mV either in the absence or presence of Co2+. Therefore, the depression of IK(full) is due to an equivalent reduction in the number of functional channels. It is concluded that Co2+ depressed IK through multiple mechanisms.

scholarly journals Measurement of calcium transients and slow calcium current in myotubes.

1994 ◽  
Vol 103 (1) ◽  
pp. 107-123 ◽  
Author(s):  
J García ◽  
K G Beam
Keyword(s):  
Skeletal Muscle ◽  
Calcium Current ◽  
Calcium Transient ◽  
Calcium Currents ◽  
Calcium Transients ◽  
Patch Clamp Technique ◽  
Slow Increase ◽  
Adult Skeletal Muscle ◽  
Calcium Indicator ◽  
Cultured Myotubes

The purpose of this study was to characterize excitation-contraction (e-c) coupling in myotubes for comparison with e-c coupling of adult skeletal muscle. The whole cell configuration of the patch clamp technique was used in conjunction with the calcium indicator dye Fluo-3 to study the calcium transients and slow calcium currents elicited by voltage clamp pulses in cultured myotubes obtained from neonatal mice. Cells were held at -80 mV and stimulated with 15-20 ms test depolarizations preceded and followed by voltage steps designed to isolate the slow calcium current. The slow calcium current had a threshold for activation of about 0 mV; the peak amplitude of the current reached a maximum at 30 to 40 mV a and then declined for still stronger depolarizations. The calcium transient had a threshold of about -10 mV, and its amplitude increased as a sigmoidal function of test potential and did not decrease again even for test depolarizations sufficiently strong (> or = 50 mV) that the amplitude of the slow calcium current became very small. Thus, the slow calcium current in myotubes appears to have a negligible role in the process of depolarization-induced release of intracellular calcium and this process in myotubes is essentially like that in adult skeletal muscle. After repolarization, however, the decay of the calcium transient in myotubes was very slow (hundreds of ms) compared to adult muscle, particularly after strong depolarizations that triggered larger calcium transients. Moreover, when cells were repolarized after strong depolarizations, the transient typically continued to increase slowly for up to several tens of ms before the onset of decay. This continued increase after repolarization was abolished by the addition of 5 mM BAPTA to the patch pipette although the rapid depolarization-induced release was not, suggesting that the slow increase might be a regenerative response triggered by the depolarization-induced release of calcium. The addition of either 0.5 mM Cd2+ + 0.1 mM La3+ or the dihydropyridine (+)-PN 200-110 (1 microM) reduced the amplitude of the calcium transient by mechanisms that appeared to be unrelated to the block of current that these agents produce. In the majority of cells, the decay of the transient was accelerated by the addition of the heavy metals or the dihydropyridine, consistent with the idea that the removal system becomes saturated for large calcium releases and becomes more efficient when the size of the release is reduced.

Voltage-dependent calcium currents in rat gonadotropes separated by centrifugal elutriation

1990 ◽  
Vol 258 (4) ◽  
pp. E589-E596 ◽  
Author(s):  
C. Marchetti ◽  
G. V. Childs ◽  
A. M. Brown
Keyword(s):  
Time Constant ◽  
Calcium Current ◽  
Low Voltage ◽  
Sodium Current ◽  
Ionic Currents ◽  
Physiological Solution ◽  
Calcium Currents ◽  
Pituitary Cells ◽  
Centrifugal Elutriation ◽  
Rat Pituitary Cells

Centrifugal elutriation of rat pituitary cells yielded two functionally active fractions (6 and 7) that contained 30-40 and 50-60% gonadotropes, respectively. We studied the membrane ionic currents of these cells with the whole cell patch-clamp method. In physiological solution, cells from the enriched fractions displayed a tetrodotoxin-sensitive sodium current. In 20 mM external calcium and zero sodium, the inward current contained two components that were different in threshold, steady-state inactivation, and deactivation. One component was half activated at approximately -25 mV, half inactivated from a holding potential of -61 mV, and deactivated with a time constant of 3 ms. The second component was half activated at 6 mV and deactivated with a time constant of 0.35 ms. These two currents resembled the high-voltage-activated and low-voltage-activated calcium current described in many preparations, including clonal and primary pituitary cells. Bath application of 20 nM gonadotropin-releasing hormone caused a transient and reversible decrease of the calcium current at depolarized voltages and a negative shift of 10 mV in the activation curve. Both effects were observed in a percentage of cells that closely matches the percentage of gonadotropes in the fraction. The shift appeared to affect both components of the current and can partially account for the increased activity of Ca2+ channels at potentials close to the resting value.

scholarly journals Spatial Distribution and Characteristics of Voltage-Gated Calcium Signals Within Visual Interneurons

2000 ◽  
Vol 83 (2) ◽  
pp. 1039-1051 ◽  
Author(s):  
Juergen Haag ◽  
Alexander Borst
Keyword(s):  
Time Constant ◽  
Voltage Dependence ◽  
Low Voltage ◽  
Cell Types ◽  
Optical Recording ◽  
Calcium Currents ◽  
Voltage Range ◽  
Spatially Resolved ◽  
Electrode Voltage

Most of our knowledge about insect calcium currents is derived from studies on cultured or dissociated somata. So far, only little data on calcium currents are available for neurons including their dendritic and presynaptic structures. Here we combined the switched-electrode voltage-clamp technique with optical recording using calcium-sensitive dyes in identified fly visual interneurons in vivo to characterize the voltage dependence and dynamics of calcium currents quantitatively and in a spatially resolved way. For all three cell types considered, i.e., centrifugal horizontal (CH), horizontal system (HS), and vertical system (VS) cells, the activation curve is rather flat and covers a voltage range from −60 to −20 mV in dendritic as well as presynaptic areas of the cells. The calcium increase is fastest for CH cells with a time constant of ∼70 ms. In HS and VS cells, the time constant amounts to 400–700 ms. The calcium dynamics as determined in different regions of the cells are similar except for a small segment between the axon and the dendrite in HS and VS cells, where the calcium increase is significantly faster. In summary, the results show the existence of a low-voltage-activated calcium current with little or no inactivation in dendritic as well as presynaptic regions of fly lobula plate tangential cells.

Postnatal Development Differentially Affects Voltage-Activated Calcium Currents in Respiratory Rhythmic Versus Nonrhythmic Neurons of the Pre-Bötzinger Complex

2005 ◽  
Vol 94 (2) ◽  
pp. 1423-1431 ◽  
Author(s):  
Frank P. Elsen ◽  
Jan-Marino Ramirez
Keyword(s):  
Postnatal Development ◽  
Calcium Current ◽  
Respiratory Rhythm ◽  
Calcium Currents ◽  
Postnatal Life ◽  
Rhythm Generation ◽  
Patch Clamp Technique ◽  
Early Postnatal Development ◽  
Bötzinger Complex ◽  
Respiratory Rhythm Generation

The mammalian respiratory network reorganizes during early postnatal life. We characterized the postnatal developmental changes of calcium currents in neurons of the pre-Bötzinger complex (pBC), the presumed site for respiratory rhythm generation. The pBC contains not only respiratory rhythmic (R) but also nonrhythmic neurons (nR). Both types of neurons express low- and high-voltage-activated (LVA and HVA) calcium currents. This raises the interesting issue: do calcium currents of the two co-localized neuron types have similar developmental profiles? To address this issue, we used the whole cell patch-clamp technique to compare in transverse slices of mice LVA and HVA calcium current amplitudes of the two neuron populations (R and nR) during the first and second postnatal week (P0–P16). The amplitude of HVA currents did not significantly change in R pBC-neurons (P0–P16), but it significantly increased in nR pBC-neurons during P8–P16. The dehydropyridine (DHP)-sensitive current amplitudes did not significantly change during the early postnatal development, suggesting that the observed amplitude changes in nR pBC-neurons are caused by (DHP) insensitive calcium currents. The ratio between HVA calcium current amplitudes dramatically changed during early postnatal development: At P0–P3, current amplitudes were significantly larger in R pBC-neurons, whereas at P8–P16, current amplitudes were significantly larger in nR pBC-neurons. Our results suggest that calcium currents in pBC neurons are differentially altered during postnatal development and that R pBC-neurons have fully expressed calcium currents early during postnatal development. This may be critical for stable respiratory rhythm generation in the underlying rhythm generating network.

Agonists for neuropeptide Y receptor subtypes NPY-1 and NPY-2 have opposite actions on rat nodose neuron calcium currents

1993 ◽  
Vol 70 (1) ◽  
pp. 324-330 ◽  
Author(s):  
J. W. Wiley ◽  
R. A. Gross ◽  
R. L. MacDonald
Keyword(s):  
Neuropeptide Y ◽  
Calcium Current ◽  
Calcium Currents ◽  
Peak Amplitude ◽  
High Threshold ◽  
Receptor Subtypes ◽  
Patch Clamp Technique ◽  
Voltage Dependent ◽  
Low Threshold ◽  
Ganglion Neurons

1. The whole-cell variation of the patch-clamp technique was used to study the effect of neuropeptide Y (NPY) and preferential agonists for the NPY-1 and NPY-2 receptor subtypes on voltage-dependent calcium currents in acutely dissociated postnatal rat nodose ganglion neurons. 2. Both low- and high-threshold calcium current components were present. NPY altered voltage-dependent calcium currents in approximately 50% of neurons studied. NPY (0.1-100 nM, ED50 6 nM) decreased the peak amplitude of transient high-threshold calcium currents in approximately 45% of the neurons. NPY (100 nM) decreased the peak amplitude of these currents 31 +/- 5% (mean +/- SE). However, in approximately 5% of the neurons NPY (100 nM) caused a reversible and reproducible increase in transient high-threshold calcium currents of 21 +/- 4%. NPY did not affect either transient low-threshold or slowly inactivating high-threshold calcium current components. 3. Application of the C-terminal fragment NPY 13-36 (100 nM), a preferential agonist for NPY-2 receptors, reversibly decreased the peak amplitude of transient high-threshold calcium currents by 26 +/- 5% in 9 of 20 cells (45%). Application of [Pro34]-NPY (100 nM), a preferential agonist for NPY-1 receptors, reversibly increased the peak amplitude of transient high-threshold calcium currents 20 +/- 4% in 23 out of 48 neurons (48%). Six of 20 neurons (30%) responded to application of both agonists. Neither the NPY-1 nor NPY-2 agonists affected transient low-threshold or slowly inactivating high-threshold calcium current components.(ABSTRACT TRUNCATED AT 250 WORDS)

Voltage-Dependent Calcium Currents in Bulbospinal Neurons of Neonatal Rat Rostral Ventrolateral Medulla: Modulation by α2-Adrenergic Receptors

1998 ◽  
Vol 79 (2) ◽  
pp. 583-594 ◽  
Author(s):  
Yu-Wen Li ◽  
Patrice G. Guyenet ◽  
Douglas A. Bayliss
Keyword(s):  
Adrenergic Receptors ◽  
Calcium Current ◽  
Low Voltage ◽  
Lucifer Yellow ◽  
Inhibitory Effect ◽  
Rostral Ventrolateral Medulla ◽  
Calcium Currents ◽  
Neonatal Rat ◽  
Ventrolateral Medulla ◽  
Voltage Dependent

Li, Yu-Wen, Patrice G. Guyenet, and Douglas A. Bayliss. Voltage-dependent calcium currents in bulbospinal neurons of neonatal rat rostral ventrolateral medulla: modulation by α2-adrenergic receptors. J. Neurophysiol. 79: 583–594, 1998. The properties and modulation by norepinephrine (NE) of voltage-dependent calcium currents were studied in bulbospinal neurons ( n = 116) of the rostral ventrolateral medulla (RVLM) using whole cell patch-clamp techniques in neonatal rat brain stem slices. RVLM bulbospinal neurons were identified visually by their location in slices and by the presence of flourescein isothiocyanate-tagged microbeads, which were injected into the spinal cord before the experiment; RVLM neurons were filled with Lucifer yellow during recordings, and the slice was processed for detection of tyrosine hydroxylase immunoreactivity (TH-IR). Thirty-four of 42 recovered cells (81%) were positive for TH-IR, indicating that most recorded cells were C1 neurons. Bulbospinal RVLM neurons expressed a prominent high-voltage–activated (HVA) calcium current, which began to activate at −30 to −40 mV (from a holding potential of −60 or −70 mV), and peaked at ∼0 mV (0.8 ± 0.1 nA;mean ± SE). HVA current comprised predominantly ω-conotoxin GVIA-sensitive, N-type and ω-agatoxin IVA-sensitive, P/Q-type components, with smaller dihydropyridine-sensitive, L-type, and residual current components. Most RVLM bulbospinal neurons ( n = 44/52, including 12/14 histologically identified C1 cells) also expressed low-voltage–activated (LVA) calcium current. LVA current began to activate at ∼−60 mV (from a holding potential of −100 mV) and was nearly completely inactivated at −50 mV with a half-inactivation potential of −70 ± 2 mV. The amplitude of LVA current at −50 mV was 78 ± 24 pA with Ba2+ and 156 ± 38 pA with Ca2+ as a charge carrier. NE inhibited HVA current in most bulbospinal RVLM neurons ( n = 70/77) with an EC50 of 1.2 μM; NE had no effect on LVA current. Calcium current inhibition by NE was mediated by α2-adrenergic receptors (α2-ARs) as the effect was mimicked by the selective α2-AR agonist, UK-14,304, and blocked by idazoxan, an α2-AR antagonist, but unaffected by prazosin and propranolol (α1- and β-AR antagonists, respectively). Most of the NE-sensitive calcium current was N- and P/Q-type. NE-induced inhibition of calcium current evoked by action potential waveforms (APWs) was significantly larger than that evoked by depolarizing steps (34 ± 2.5 vs. 23 ± 2.7%; P < 0.05). Although inhibition of calcium current was voltage dependent and partially relieved by strong depolarizations, when calcium currents were evoked with a 10-Hz train of APWs as a voltage command, the inhibitory effect of NE was maintained throughout the train. In conclusion, bulbospinal RVLM neurons, including C1 cells, express multiple types of calcium currents. Inhibition of HVA calcium current by NE may modulate input-output relationships and release of transmitters from C1 cells.

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