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Biomedical subjects

D Attwell

Publications and source records attributed to D Attwell.

At least 73 records · Page 4Linked to original sources
PubMed1988

Membrane currents in retinal bipolar cells of the axolotl.

By whole-cell patch-clamping bipolar cells isolated from enzymatically dissociated retinae, we have studied the nonsynaptic ionic currents that may play a role in shaping the bipolar cell light response and in determining the level of voltage noise in these cells. Between -30 and -70 mV, the membrane current of isolated bipolar cells is time independent, and the input resistance is 1-2 G omega. Depolarization past -30 mV activates an outward current (in less than 100 ms), which then inactivates slowly (approximately 1 s). Inactivation of this current is removed by hyperpolarization over the range -20 to -80 mV. This current is carried largely by K ions. It is not activated by internal Ca2+. The membrane current of isolated bipolar cells is noisy, and the variance of this noise has a minimum between -40 and -60 mV. At its minimum, the standard deviation of the voltage noise produced by nonsynaptic membrane currents is at least 100 microV. The membrane currents of depolarizing bipolar cells in slices of retina were investigated by whole-cell patch-clamping. Their membrane properties were similar to those of isolated bipolar cells, but with a larger membrane capacitance and a smaller input resistance. Their membrane current noise also showed a minimum near -40 to -60 mV. The time-dependent potassium current in axolotl bipolar cells is not significantly activated in the physiological potential range and can therefore play little role in shaping the bipolar cells' voltage response to light. Differences in the waveform of the light response of bipolar cells and photoreceptors must be ascribed to shaping by the synapses between these cells. The noise minimum in the bipolar membrane current is near the dark potential of these cells, and this may be advantageous for the detection of weak signals by the bipolar cells.

Adenosine Triphosphate↗
PubMed1987

Noise analysis predicts at least four states for channels closed by glutamate.

For ion channels that are opened by neurotransmitters, analysis of current noise has given valuable information on the kinetics of synaptic channel gating. In depolarizing bipolar cells of the vertebrate retina, we have recently characterized a synaptic current for which the neurotransmitter glutamate closes channels, and for which the channel open probability is low even in the absence of glutamate. We present here predictions for the current noise spectrum expected for various models of glutamate's action on the ion channels. Comparison of these theoretical predictions with experimental data allows us to rule out several simple kinetic schemes for the action of glutamate, and to conclude that the channels closed by glutamate must be able to exist in at least four different states.

Animals↗
PubMed1987

Neurotransmitter-induced currents in retinal bipolar cells of the axolotl, Ambystoma mexicanum.

1. Whole-cell patch clamping was used to study the membrane properties of isolated bipolar cells and the currents evoked in them by putative retinal neurotransmitters. 2. Isolated bipolar cells show an approximately ohmic response to voltage steps over most of the physiological response range, with an average input resistance of 1.3 G omega and resting potential of -35 mV. These values are underestimates because of the shunting effect of the seal between the patch electrode and the cell membrane. Depolarization beyond -30 mV produces rapid activation (10-100 ms) of an outward current (carried largely by potassium ions), which then inactivates slowly (0.5-2 s). 3. Of five candidates for the photoreceptor transmitter, four (aspartate, N-acetylhistidine, cadaverine, putrescine) had no effect on bipolar cells. The fifth substance, L-glutamate, opened ionic channels with a mean reversal potential of -12 mV in some cells (presumed hyperpolarizing bipolar cells), and closed channels with a mean reversal potential of -13 mV in other cells (presumed depolarizing bipolar cells). 4. The conductance increase induced by glutamate in presumed hyperpolarizing bipolar cells was associated with an increase in membrane current noise. Noise analysis suggested a single-channel conductance for the glutamate-gated channel of 5.4 pS. The power spectrum of the noise increase required the sum of two Lorentzian curves to fit it, suggesting that the channel can exist in three states. 5. The conductance decrease induced by glutamate in presumed depolarizing bipolar cells was associated with a decrease in membrane current noise that could be described as the sum of two Lorentzian spectra, and which suggested a single-channel conductance of 11 pS. The noise decrease implies that the channels closed by glutamate are not all open in the absence of the transmitter. 6. GABA (gamma-aminobutyric acid) and glycine, transmitters believed to mediate lateral inhibition in the retina, open chloride channels in isolated bipolar cells, and increase the membrane current noise. Noise analysis suggested that the channels gated by GABA and glycine have conductances of 4.4 and 7.5 pS respectively. The noise spectra required the sum of two Lorentzian curves to fit them. 7. By whole-cell patch clamping cells in retinal slices, the synaptic transmitter released by photoreceptors was shown to close channels with an extrapolated reversal potential around -3 mV in depolarizing bipolar cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗
PubMed1985

The effect of light on the spread of signals through the rod network of the salamander retina.

Adjacent rods in the amphibian retina are electrically coupled to each other by gap junctions. By injecting current pulses into one rod and recording the voltage change produced in nearby rods, we have studied the extent to which signals spread between rods in the presence and absence of illumination. Light has little effect on the steady potentials produced in nearby rods by the injection of a hyperpolarizing current, but does affect the propagation of transient signals through the rod network. The responses to injection of depolarizing current are increased by light. These effects of light were mimicked by hyperpolarizing the rod network (non-uniformly) by injecting continuous current (on top of which current pulses were superimposed to monitor signal spread). This suggests that the effects of light are due solely to the rod hyperpolarization produced by light. The effects of light are not completely predicted from computer simulations based on a previous characterization of the properties of isolated rods; these experiments thus reveal an inadequacy in the description of the rod membrane currents in that model. Light-induced hyperpolarization of cones has no effect on signal spread between rods. The functional significance of these results is discussed.

Ambystoma↗
PubMed1985

Kinetics of light-sensitive channels in vertebrate photoreceptors.

We have studied the ion channels mediating the light response of vertebrate rod photoreceptors by analysing fluctuations in the current across the rod membrane, using the whole cell patch-clamp technique on rods isolated from the axolotl retina. Light decreases the membrane current fluctuations. Noise analysis reveals two components to this decrease: a low frequency component due to biochemical noise in the transduction mechanism, and a high frequency component we attribute to the random opening and closing of the ion channels in the dark. The probability of any one channel being open in the dark is low. The spectrum of the high frequency component of the current fluctuations indicates that the current through an open channel is 4 X 10(-15)A, that the mean channel open time is 2 ms, and that about 10000 channels are open in each rod in the dark. The effect of light is to reduce the opening rate constant of these channels, with no effect on the closing rate constant.

Ambystoma mexicanum↗
PubMed1985

Is the potassium channel distribution in glial cells optimal for spatial buffering of potassium?

Glial cells in the nervous system are believed to reduce changes of extracellular potassium concentration ([K+]o), caused by neural activity, by carrying out spatial buffering of potassium. In the case of retinal glial cells (Müller cells), light-evoked increases of [K+]o within the retina are reduced by K ions flowing through the Müller cell to the vitreous fluid of the eye. We have calculated the optimal way to distribute the potassium conductance of the Müller cell to maximize spatial buffering to the vitreous fluid. The best distribution is with half the potassium conductance in the outer part of the cell, where K+ enters, and half the conductance in the vitreal endfoot, where K+ leaves the cell. This calculated distribution is very different from the actual distribution measured by Newman (1984, Nature [Lond.], 309: 155-157), where only 6% of the Müller cell conductance is in the outer cell and 94% is in the endfoot. The experimentally observed distribution gives less than a quarter of the spatial buffering that would be produced by the optimal distribution. The possible advantages of this arrangement are discussed.

Animals↗
PubMed1985

Lead and mercury toxicity and the rod light response.

Lead and mercury have been reported to alter selectively the rod component of the electroretinogram, and to inhibit the phosphodiesterase in rod outer segments which may be responsible for generating the rods' light response. The authors have investigated the effect of lead and mercury on the voltage response to light of rods, and compared these effects with those of the phosphodiesterase inhibitor papaverine. Lead and mercury, like papaverine, slow the light response. In addition, papaverine increases the light response amplitude while lead decreases it. Mercury initially increases and then decreases the amplitude. The late decrease in amplitude produced by mercury is associated with rod degeneration: an effect which may mimic degenerative diseases in which the rod phosphodiesterase is insufficiently active. These results demonstrate that the changes of electroretinogram induced by lead and mercury can be accounted for by the changes in receptor potential these heavy metals produce. The changes in receptor potential seen are consistent with mercury inhibiting the rod phosphodiesterase, and with lead having an action in addition to phosphodiesterase inhibition.

Ambystoma↗
PubMed1984

Membrane potential affects photocurrent kinetics in salamander rods and cones.

In both rod and cone photoreceptors, polarization of the cell to positive potentials produces a slowing of the photocurrent. This could occur because, at positive potentials, the rate of removal of internal transmitter is slowed, or because the time for which the channels stay closed after interaction with internal transmitter is prolonged. Ion concentration changes within the cell may also contribute to the time course of the photocurrent.

Ambystoma↗
PubMed1984

A quantitative analysis of interactions between photoreceptors in the salamander (Ambystoma) retina.

A quantitative description of the electrical properties of the photoreceptor layer in the salamander retina was obtained from earlier data on the characteristics of isolated rods and cones and on rod-rod coupling, and from new data on rod-cone and cone-cone coupling and on the rod photocurrent. Injecting -1 nA current into a rod elicits hyperpolarizations of about 20 mV in an adjacent rod and 4 mV in an adjacent cone. Responses of more distant receptors are smaller. Injecting -1 nA into a cone elicits hyperpolarizations of about 4 mV in an adjacent rod and 0.4 mV in a nearby cone. Depolarizing current evokes smaller responses. Assuming, in agreement with anatomical evidence, that each rod is electrically coupled to four rods and to four cones around it, and that there is no direct electrical coupling between cones, we found these results could be predicted from the properties of isolated rods and cones if adjacent rods are coupled by a resistance of 300 M omega and adjacent rods and cones are coupled by a resistance of 5000 M omega. The small cone-cone coupling seen is due to coupling via intervening rods. The two halves of double cones are not electrically coupled. The spectral sensitivity of both halves is a maximum around 620 nm wave-length. The rod photocurrent has been characterized by voltage-clamping rods isolated from the retina. In agreement with Bader, MacLeish & Schwartz (1979) we found the time course of the photocurrent to be approximately independent of voltage between -35 and -85 mV. The voltage responses of rods, single cones and double cones isolated from the retina obey the principle of univariance. Responses of receptors in the retina do not obey univariance. The main deviations from univariance observed can be explained if adjacent rods and cones are coupled by a resistance of 5000 M omega. Our data demonstrate that rod-cone coupling is relatively weak. We simplified our description of the photoreceptor network, by omitting cones, to investigate the spatiotemporal processing that the rod network is capable of. Computer simulations predict, as is found experimentally, that the rod voltage response to a large spot of bright light should show a much more pronounced initial transient hyperpolarization than the response to a small spot of light of the same intensity. This difference is produced by the combination of electrical coupling of the rods with the existence of a voltage-gated current, IA, in the rod membrane.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗
PubMed1983

The spatial frequency sensitivity of bipolar cells.

Retinal bipolar cells constitute the output stage of the outer layer of the retina. There are several constraints on the ability of the bipolar cell array to respond to the different spatial frequency components of the visual image, including (i) electrical coupling in the dendritic tree receiving receptor input; (iii) the "lateral inhibition" mediated by horizontal cells. Using simple mathematical models, we derive analytical expressions for the spatial frequency response of the bipolar cell array for the case in which horizontal cells are presynaptic to bipolar cells (feedforward model) and also for the case in which horizontal cells are presynaptic to receptors (feedback model). The results illustrate the importance of the three factors mentioned in determining the bipolar cells' properties. The optimal spatial frequency for stimulating the bipolar cell array, and the range of spatial frequencies transmitted onward to the inner plexiform layer, are thus related to the anatomical and electrical properties of the cells in the outer plexiform layer.

Animals↗
PubMed1983

A new method for obtaining isolated photoreceptors from the amphibian retina.

A new method is described for obtaining isolated rods and cones by physical dissociation of the retina. The method gives a large yield of isolated photoreceptors, without any changes in membrane properties that enzymatic dissociation techniques might produce. Photovoltages and photocurrents recorded from rods isolated in this way are presented.

Ambystoma↗
PubMed1983

A sign-reversing pathway from rods to double and single cones in the retina of the tiger salamander.

Signal transmission between rods and cones was studied by passing current into a rod and recording the voltage response in a nearby double or single cone and vice versa. Two types of rod-cone interaction were found. Between immediately adjacent rods and cones, passage of current into either receptor elicited in the other receptor a sustained voltage response of the same sign as the injected current. These signals were still seen in the presence of Co2+, and are probably mediated by the electrical synapses which have been seen anatomically between adjacent rods and cones. In addition to this short-range sign-preserving interaction, passing current into a rod elicited a transient sign-inverted signal in cones up to at least 80 micron from the injected rod. No such response was seen in rods for current injection into cones. This signal was greatly reduced by Co2+ ions. Hyperpolarization of the cone to about -65 mV, with about 0.1 nA current, reversed this signal, which is presumed to be mediated by a chemical synaptic input to cones. Light flashes suppressed the sign-inverted signal for a period which was longer for brighter flashes. The time of reappearance of the signal was correlated with the return of the rod and horizontal cell potentials to their dark levels. This suppression could also be produced by an annulus of light which produced no light response in the receptors at the centre of the annulus, but which did polarize horizontal cells under the centre of the annulus. The wave form of the sign-inverted signal was similar to that produced in horizontal cells by current injection into rods, but of opposite sign. If an electrode was left in a cone for some time, the normal hyperpolarizing light response diminished, leaving a depolarizing response produced, presumably, by feed-back from horizontal cells. This signal was reversed when the cone was hyperpolarized with about 0.1 nA current. These data suggest that the sign-inverted response is mediated by feed-back from horizontal cells and, assuming that depolarization increases the rate of release of horizontal cell synaptic transmitter, then the feed-back transmitter opens channels in the cone membrane whose currents have a reversal potential around -65 mV.

Action Potentials↗
PubMed1982

The dependence on heart rate of the human ventricular action potential duration.

For the human electrocardiogram, the relationship between QT interval and heart rate was examined in conditions where the heart rate was changed at different rates. The QT interval provides a measure of the ventricular action potential duration. For slow changes of heart rate, elicited by mild exercise, the dependence of QT interval on heart rate obtained when the heart rate was rising was the same as that obtained when the heart rate was falling. The QT interval was approximately proportional to 1/(heart rate)1/2. For the rapid transient changes of heart rate associated with respiration, the QT interval was approximately constant, independent of the instantaneous heart rate. For rapid sustained changes of heart rate, elicited by heavy exercise or immersion of the face in cold water, the changes in QT interval lagged behind the heart rate changes. Consequently the dependence of QT interval on heart rate obtained when the heart rate was increasing differed from the dependence obtained during a fall in heart rate. This time dependence of the relationship between QT interval and heart rate is similar to that seen for the relationship between action potential duration and stimulation rate in isolated pieces of mammalian ventricle.

Action Potentials↗
PubMed1982

The properties of single cones isolated from the tiger salamander retina.

1. The properties of isolated single cones were studied using the voltage-clamp technique, with two micro-electrodes inserted under visual control.2. Single cones had input resistances, when impaled with two electrodes, of up to 270 MOmega. This is probably lower than the true membrane resistance, because of damage by the impaling electrodes. The cone capacitance was about 85 pF.3. The cone membrane contains a time-dependent current, I(B), controlled by voltage, and a separate photosensitive current.4. The gated current, I(B), is an inward current with a reversal potential around -25 mV. It is activated by hyperpolarization over the range -30 to -80 mV, and at constant voltage obeys first order (exponential) kinetics. The gating time constant is typically 50 ms at the resting potential of -45 mV, rises to 170 ms at -70 mV, and decreases for further hyperpolarization.5. The spectral sensitivity curve of the cone light response peaks at 620 nm wave-length, and is narrower than the nomogram for vitamin A(2)-based pigments. The light responses of isolated cones are spectrally univariant.6. Voltage-clamped photocurrents were recorded at various membrane potentials, for light steps of various intensities. The photocurrent reversed at around -8 mV. The time course of the photocurrent, for a given intensity, was approximately independent of voltage (although its magnitude was voltage-dependent). The shape of the peak current-voltage relation of the light-sensitive current was independent of light intensity (although its magnitude was intensity-dependent).7. These results can be explained if: (a) light simply changes the number of photosensitive channels open, without altering the properties of an open channel; (b) the reactions controlling the production of internal transmitter, the binding of internal transmitter to the photosensitive channels, and the closing and opening of the channels are unaffected by the electric field in the cone membrane, even though at least some of these reactions take place in the membrane.8. I(B) plays only a small role in shaping the cone voltage response to light.

Ambystoma↗
PubMed1981

The dependence of the maximum rate of rise of the action potential upstroke on membrane properties.

The maximum rate of rise of the action potential (Vmax) is often used to study the maximum sodium conductance (GNa) of excitable membranes, by assuming that Vmax is proportional to GNa. However, the real relation between Vmax and GNa is uncertain. We use simple analytical descriptions of the membrane currents to investigate this relation. If (1) the sodium conductance is greater than the non-sodium conductance of the membrane, (2) the sodium current is activated extremely quickly, and (3) the sodium current is inactivated extremely slowly, then Vmax will indeed be proportional to GNa. However, if conditions (1) or (3) are not satisfied, the Vmax-GNa relation will be non-proportional, such that a certain fractional change of GNa produces a larger fractional change of Vmax. If condition (2) is not satisfied the Vmax-GNa relation is distorted in the opposite direction, such that a certain fractional change of GNa produces a smaller fractional change of Vmax. Measurements of Vmax are usually performed in preparations where voltage clamping cannot be used to study GNa directly. However, voltage clamping is necessary to verify that conditions (1)-(3) are satisfied. The results of studies using Vmax alone as a measure of GNa should be assessed with caution.

Action Potentials↗
PubMed1981

The timing of channel opening during miniature end-plate currents.

Many chemical transmitters act by opening channels with exponentially distributed life-times. We present a way to analyze a synaptic current in terms of its component channels. We can estimate the numbers and times of channel opening within the synaptic current. This approach is used to study miniature end-plate currents (m.e.p.c.s) at the frog neuromuscular junction. The results support the idea that some transmitter rebinds after dissociation from post-synaptic receptors, and suggest that the time of channel closing is related to the time at which acetylcholine leaves the receptor.

Acetylcholine↗