Visual adaptation modulates a potassium conductance in retinular cells of the crayfish

2000 ◽  
Vol 17 (3) ◽  
pp. 353-368 ◽  
Author(s):  
C.S. MILLER ◽  
R.M. GLANTZ

Crayfish photoreceptors exhibit a voltage-dependent potassium conductance, GK, that is generally similar to the delayed rectifier channel described in neurons and other arthropod retinular cells. GK activation (i.e. the apparent threshold, Vth) occurs near the resting potential and GK is substantially reduced by 25 mM extracellular tetraethylammonium (TEA) and by intracellular Cs+ injections. Light exposure, sufficient to reduce visual sensitivity 100-fold, increases Vth (shifts it in the depolarizing direction) by about 20 mV. The light-dependent change in Vth does not depend upon the corresponding increase (depolarization) of the steady-state membrane potential nor does it depend upon inward calcium currents. Vth is slightly influenced by fluctuations in Ko associated with the light-elicited currents. During light exposure Ko (measured with K+-sensitive electrodes) increases by 2.1 mM (equivalent to an 8 mV increase in EK). This increase in EK makes only a modest contribution to the light-dependent change in Vth as determined by perfusion with high potassium salines. Intracellular calcium injections increase Vth by 10 to 20 mV and reduce visual sensitivity by 5- to 10-fold. The results imply that during exposure to high levels of illumination, K+ currents at the steady-state membrane potential are diminished by a calcium-dependent change in GK gating and, to a smaller degree, by a reduced K+ concentration gradient. It is notable that Ca2+ appears to inhibit both GK and the light-elicited conductance from both inside and outside the plasma membrane. As a consequence of the light-dependent change in Vth, GK makes only modest contributions to the changes in sensitivity and speed normally associated with light adaption. These functions are regulated by the transduction pathway and are revealed at the resting potential in the time course and magnitude of the light-elicited currents.

1959 ◽  
Vol 196 (5) ◽  
pp. 1071-1078 ◽  
Author(s):  
Robert E. Taylor

Procaine (0.025–0.1%; pH 7.9) caused a reduction in the amount and rate of development of the early transient (sodium) and late steady state (potassium) currents which occur during a depolarizing voltage step applied to the excised, voltage clamped squid axon. Consistent results were obtained by holding the membrane potential at a hyperpolarized value prior to the applied step. No effect was seen on the resting potential, on the sodium equilibrium potential, or on the proportion of the sodium carrying system which was ‘inactive’ at any membrane potential. The blocking action of procaine is a result of the inhibition by the drug of the sodium carrying system. The effect of procaine on the potassium conductance is such as to oppose the blocking action.


1989 ◽  
Vol 62 (6) ◽  
pp. 1280-1286 ◽  
Author(s):  
N. Uchimura ◽  
E. Cherubini ◽  
R. A. North

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.


1980 ◽  
Vol 58 (9) ◽  
pp. 999-1010 ◽  
Author(s):  
Peter C. Vaughan ◽  
James G. McLarnon ◽  
Donald D. F. Loo

Three-microelectrode voltage-clamp experiments have been conducted on surface fibres of Xenopus laevis sartorius muscles. When potassium and chloride were substituted by rubidium and sulphate, negligibly small currents were observed. In solutions containing rubidium and chloride at pH 8.4–8.8 normally polarized fibres exhibited instantaneous current–voltage relations that were linear over a wide voltage range. Chloride conductance varied widely from fibre to fibre; the mean resting conductance at −80 mV was 7.4 × 10−4 ± 2.6 × 10−4 S/cm2 (mean ± SE). When hyperpolarizing voltage steps were made, conductance declined from the initial to the steady state; inward currents saturated near 14 μA/cm2. In experiments performed on fibres depolarized by immersion in K+-and Rb+-rich solutions it was found that resting conductance did not increase by as much as would be expected from constant field – constant permeability precepts, by comparison with normally polarized fibres. Despite the low chloride transmembrane concentration ratio, rectification in the steady state was similar in depolarized and normally polarized fibres.When a two-pulse protocol was employed to test the availability of chloride conductance after conditioning of the system at some voltage, it was found that the test current, the initial current at the onset of the test voltage step, depended sigmoidally on the conditioning voltage. The sigmoid relationships had asymptotic limits: after hyperpolarizing conditioning the test current was minimal, after depolarizing conditioning, maximal. Normalized sigmoid relations were superimposable, whether from normally polarized or chronically depolarized cells.When the protocol was repeated using different test potentials and initial currents following a particular conditioning voltage were plotted against the test potential, families of straight lines were obtained. The slopes of the members of these families were dependent on the conditioning voltage: the more negative the conditioning step the lower the slope. The lines projected through a mutual intersection at a voltage slightly more positive than the resting potential. This is interpreted as indicating that there is some voltage, slightly positive with respect to the membrane potential, at which the initial current is independent of the conditioning voltage.It is concluded that the state of the chloride conductance mechanism is a function of the deviation of the membrane from the resting potential rather than of the absolute membrane potential and that relaxations from initial to steady states reflect properties of the permeation mechanism rather than accumulation or depletion of chloride in a confined space, although some contribution by a mechanism such as the latter cannot be completely ruled out.


1988 ◽  
Vol 66 (10) ◽  
pp. 1328-1336 ◽  
Author(s):  
C. Krieger ◽  
T. A. Sears

The development of voltage-dependent ionic conductances of foetal mouse spinal cord neurones was examined using the whole-cell patch-clamp technique on neurones cultured from embryos aged 10–12 days (E10–E12) which were studied between the first day in vitro (V1) to V10. A delayed rectifier potassium conductance (IK) and a leak conductance were observed in neurones of E10.V1, E11, V1, and E12, V1 as well as in neurones cultured for longer periods. A rapidly activating and inactivating potassium conductance (IA) was seen in neurones from E11, V2 and E12, V1 and at longer times in vitro. A tetrodotoxin (TTX) sensitive sodium-dependent inward current was observed in neurones of E11 and E12 from V1 onwards. Calcium-dependent conductances were not detectable in these neurones unless the external calcium concentration was raised 10- to 20-foid and potassium conductances were blocked. Under these conditions calcium currents could be observed as early as E11, V3 and E12, V2 and at subsequent times in vitro. The pattern of development of voltage-dependent ionic conductances in murine spinal neurones is such that initially leak and potassium currents are present followed by sodium current and subsequently calcium current.


1959 ◽  
Vol 43 (2) ◽  
pp. 315-321 ◽  
Author(s):  
C. Edwards ◽  
S. Hagiwara

The effect of the absence of potassium in the bathing solution on the synaptic inhibitory potentials of the crayfish stretch receptor has been studied. The inhibitory potentials were increased in size, i.e. became more hyperpolarizing, in the absence of potassium. Since the resting potential of the cell is increased in the absence of potassium, the alteration of the inhibitory potentials implies that the potassium conductance of the membrane is increased. While other ions, e.g. Cl-, may also be involved, it seems that the membrane potential during inhibition is mainly dominated by K+.


2005 ◽  
Vol 17 (2) ◽  
pp. 290 ◽  
Author(s):  
R. Boni ◽  
N. Cocchia ◽  
F. Silvestre ◽  
G. Tortora ◽  
R. Lorizio ◽  
...  

The low developmental efficiency recorded in juvenile oocytes represents, besides its technological relevance, an opportunity for increasing the knowledge of mechanisms regulating developmental competence in the oocytes. To analyze the biological reasons that make an adult oocyte different from a juvenile one, we monitored membrane electrical properties, i.e. resting potential, steady-state conductance and calcium currents, and calcium stores in these two oocyte types both at immature (GV) stage and after in vitro maturation (MII). Ovaries of cycling ewes and 40-day-old lambs were collected at abattoir and transported at 30°C. Cumulus-oocyte complexes (COC) were recovered by mincing. In vitro maturation was carried out in TCM199 supplemented with 10% fetal calf serum, 10 IU/mL of LH, 0.1 IU/mL of FSH, and 1 mg/mN of 17β-estradiol at 39.0°C in 5% CO2 for 24 h. Zona pellucida of immature and in vitro-matured oocytes was removed after incubation for 1–1.5 min in 0.5% (w/v) protease solution. Zona-free oocytes were placed in Ham F10 at 38.5°C and voltage clamped by standard techniques (Tosti et al. 2002 Reproduction 124, 835–846). After obtaining a giga-seal, the patch was ruptured. The permeability of the plasma membrane was verified by applying depolarizing and hyperpolarizing voltage steps of 10 mV and 500 ms before and at the peak current to generate the voltage-dependent currents. The voltage clamp was set at −80 and −30 mV to differentiate the Ca2+ current components, i.e. L-type Ca2+ channels. For intracellular calcium determinations, oocytes were placed in Ham F10 and injected with the 0.5 mM calcium green dextran (Mr 10,000). Ca2+ stores were evoked by the addition of 5 μM Ca2+ ionophore, monitored using a computer-controlled photo-multiplier system, and measured as relative fluorescence units (RFU) by normalizing fluorescence against baseline fluorescence. In lamb and ewe, differences in electrical features and calcium dynamics between GV (n = 36 and 17) and MII (n = 42 and 32) oocytes were tested by ANOVA and expressed as mean ± SEM. Resting potential was higher at MII than GV stages (−15.2 ± 0.9 vs. −12.1 ± 1.1 mV, respectively; P < 0.02) but it did not differ between animal age. GV stage and ewe showed either a higher steady-state conductance (25.4 ± 0.2 vs. 11.7 ± 0.2 nS and 21.7 ± 0.2 vs. 15.4 ± 0.2 nS, respectively; P < 0.01) or L-type Ca2+ channels (9.7 ± 1.4 vs. 2.7 ± 1.3 pA and 9.2 ± 1.5 vs. 3.2 ± 1.1 pA, respectively; P < 0.01). No differences were found between resting potential peaks yielded after Ca2+ ionophore exposure but a higher ion activation current was found in lamb oocytes (489 ± 56 vs. 300 ± 73 pA; P < 0.05). Ca2+ stores did not differ between animal age but they were larger at MII than at GV stage (0.70 ± 0.07 vs. 0.44 ± 0.07 RFU; P < 0.01). These results supply further information on both reproductive biology in ovine species and the physiology of oocytes collected from juvenile and adult individuals. This work was supported by Italian Ministry of University and Research (MIUR) COFIN 2002 Project.


1991 ◽  
Vol 155 (1) ◽  
pp. 305-321
Author(s):  
D. A. DORSETT ◽  
C. G. EVANS

Dissociated unstriated muscle fibres from the buccal mass retractor muscles of the mollusc Philine aperta were studied using a two-electrode voltage-clamp. The mean resting potential of the fibres was −76.3±0.44mV (N=30), and the membrane resistance was 42.2±3MΩ. The space constant of the fibres was 2.03+0.33mm (N=5). Three outward potassium currents were resolved in response to a depolarising step to zero from resting potential. (1) An early transient current, voltageactivated and blocked by 2 mmoll−1 4-aminopyridine (4-AP). This resembled the A-current described in molluscan neurones and some arthropod muscle fibres. (2) A calcium-dependent late transient current, with slower kinetics, which was suppressed by 50 mmoll−1 tetraethylammonium chloride (TEA-Cl), zero-calcium saline, 1 mmol−1 Cd2+ and 1 μmoll−1 verapamil. (3) A delayed voltage-activated current, blocked by 50 mmoll−1 TEA-Cl and with kinetics associated with the delayed rectifier current IK. An inwardly directed current, blocked by zero-calcium salines, Cd2+ and verapamil, was considered to be a calcium current whose activation closely matched that of the Ca2+-dependent potassium current. A blockade of either the A-current, or exposure to low-calcium artificial sea water, or a combination of both, promoted the development of oscillations and regenerative spikes in the muscle fibre following depolarization


1995 ◽  
Vol 269 (3) ◽  
pp. F289-F308 ◽  
Author(s):  
X. Gu ◽  
H. Sackin

The effect of intracellular and extracellular pH on potassium conductance (GK) was examined in isolated amphibian (Rana pipiens) proximal tubule cells under whole cell voltage clamp conditions. Internal perfusion of the patch pipette was used to precisely control intracellular pH. In the region of normal resting potential (-51 +/- 3 mV), raising cell pH from 6.5 to 8.0 did not significantly increase GK (1.1 +/- 0.3 vs. 1.3 +/- 0.3 nS; P > 0.08, n = 8). Similar elevations in external (bath) pH had even less of an effect on GK. In contrast, when cells were voltage clamped to 30 mV more negative than the resting potential, raising internal pH from 6.5 to 8.0 did increase GK from 1.05 +/- 0.3 to 1.8 +/- 0.5 nS (P < 0.04; n = 8). These results suggest that modest changes in pH have little effect on GK, except at large negative potentials. In the process of examining the pH dependence of GK, a slowly activating, voltage-dependent conductance of 7.5 +/- 1 nS (n = 20; for 20 microns cells) was observed during cell depolarization. Although the instantaneous current-voltage relation of this conductance was linear, its marked voltage dependence produced an apparent steady-state rectification, with Gm = 0.5 +/- 0.2 nS and Gout = 9.0 +/- 1 nS (n = 11). Outward current was reversibly blocked by 3 mM Cu, Cd, or Co. In the absence of Na, K, and Ca (and only trace amounts of Cl), rapid changes in bath pH from 6.5 to 8.0 shifted the steady-state reversal potential (Erev) by -37 +/- 4 mV (n = 9) and the instantaneous Erev by -56 +/- 4 mV (n = 9). These shifts in Erev were consistent with a hydrogen ion conductance (GH), similar to what has been reported for snail neuron, neutrophils, alveolar epithelial cells, and phagocytes. Since the magnitude of this GH would be insignificant at resting cell pH and membrane potential, its role in renal proximal tubule under normal conditions is somewhat obscure. Nonetheless, in pathological situations, GH could function to prevent acid overload during any process that depolarizes the cell, such as low temperature or metabolic inhibition.


1998 ◽  
Vol 79 (1) ◽  
pp. 205-216 ◽  
Author(s):  
Hans-Christian Pape ◽  
Denis Paré ◽  
Robert B. Driesang

Pape, Hans-Christian, Denis Paré, and Robert B. Driesang. Two types of intrinsic oscillations in neurons of the lateral and basolateral nuclei of the amygdala. J. Neurophysiol. 79: 205–216, 1998. Intracellular recordings in the guinea pig and cat basolateral amygdaloid (BL) complex maintained as slices in vitro revealed that a subpopulation of neurons (79%) in the lateral (AL) and basolateral (ABl) nuclei generated two types of slow oscillations of the membrane potential upon steady depolarization from resting potential. The cells were of a stellate or pyramidal-like shape and possessed spiny dendrites and an axon leaving the local synaptic environment, and thus presumably represented projection neurons. Similar oscillatory activity was observed in projection neurons of the cat AL nucleus recorded in vivo. Oscillatory activity with a low threshold of activation (low-threshold oscillation, LTO) appeared as rhythmic deflections (amplitudes, 2–6 mV) of the membrane potential positive to −60 mV. Fast Fourier transformation (FFT) demonstrated a range of frequencies of LTOs between 0.5 and 9 Hz, with >80% occurring at 1–3.5 Hz and an average at 2.3 ± 1.1 Hz. LTOs were more regular after pharmacological blockade of synaptic transmission and were blocked by tetrodotoxin (TTX). Blockade of LTOs and Na+ spikes revealed a second type of oscillatory activity (high-threshold oscillation, HTO) at depolarizations beyond −40 mV, which was capable of triggering high-threshold spikes. HTOs ranged between 1 and 7.5 Hz, with >80% occurring at 2–6 Hz and an average at 5.8 ± 1.1 Hz. HTOs vanished at a steady membrane polarization positive to −20 mV. Current versus voltage relations obtained under voltage-clamp conditions revealed two regions of negative slope conductance at −55 to −40 mV and at around −30 mV, which largely overlapped with the voltage ranges of LTOs and HTOs. TTX abolished the first region of negative slope conductance (−55 to −40 mV) and did not significantly influence the second region of negative slope conductance. Neuronal responses to maintained depolarizing current pulses consisted of an initial high-frequency discharge (up to 100 Hz), the frequency of which depended on the amplitude of the depolarizing current pulse, followed by a progressive decline (“adaptation”) toward a slow-rhythmic firing pattern. The decay in firing frequency followed a double-exponential function, with time constants averaging 57 ± 28 ms and 3.29 ± 1.85 s, and approached steady-state frequencies at 6.3 ± 2.9 Hz ( n = 17). Slow-rhythmic firing remained at this frequency over a wide range of membrane polarization between approximately −50 and −20 mV, although individual electrogenic events changed from Na+ spikes and underlying LTOs to high-threshold spikes and underlying HTOs. Rhythmic regular firing was only interrupted at an intermediate range of membrane polarization by the occurrence of spike doublets. In conclusion, the integrative behavior of a class of neurons in the BL complex appears to be largely shaped by the slow-oscillatory properties of the membrane. While LTOs are likely to synchronize synaptic signals near firing threshold, HTOs are a major determinant for the slow steady-state firing patterns during maintained depolarizing influence. These intrinsic oscillatory mechanisms, in turn, can be assumed to promote population activity at this particular frequency, which ranges well within that of the limbic theta (Θ) rhythm and the delta (δ) waves in the electroencephalogram during slow-wave sleep.


2002 ◽  
Vol 88 (2) ◽  
pp. 869-878 ◽  
Author(s):  
Marcel de Jeu ◽  
Alwin Geurtsen ◽  
Cyriel Pennartz

A Ba2+-sensitive K+ current was studied in neurons of the suprachiasmatic nucleus (SCN) using the whole cell patch-clamp technique in acutely prepared brain slices. This Ba2+-sensitive K+ current was found in approximately 90% of the SCN neurons and was uniformly distributed across the SCN. Current-clamp studies revealed that Ba2+ (500 μM) reversibly depolarized the membrane potential by 6.7 ± 1.3 mV ( n = 22) and concomitantly Ba2+ induced an increase in the spontaneous firing rate of 0.8 ± 0.2 Hz ( n = 12). The Ba2+-evoked depolarizations did not depend on firing activity or spike dependent synaptic transmission. No significant day/night difference in the hyperpolarizing contribution to the resting membrane potential of the present Ba2+-sensitive current was observed. Voltage-clamp experiments showed that Ba2+ (500 μM) reduced a fast-activating, voltage-dependent K+ current. This current was activated at levels below firing threshold and exhibited outward rectification. The Ba2+-sensitive K+ current was strongly reduced by tetraethylammonium (TEA; 20 and 60 mM) but was insensitive to 4-aminopyridine (4-AP; 5 mM) and quinine (100 μM). A component of Ba2+-sensitive K+ current remaining in the presence of TEA exhibited no clear voltage dependence and is less likely to contribute to the resting membrane potential. The voltage dependence, kinetics and pharmacological properties of the Ba2+- and TEA-sensitive K+ current make it unlikely that this current is a delayed rectifier, Ca2+-activated K+ current, ATP-sensitive K+ current, M-current or K+ inward rectifier. Our data are consistent with the Ba2+- and TEA-sensitive K+ current in SCN neurons being an outward rectifying K+ current of a novel identity or belonging to a known family of K+ channels with related properties. Regardless of its precise molecular identity, the current appears to exert a significant hyperpolarizing effect on the resting potential of SCN neurons.


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