Hyperpolarization of a barnacle photoreceptor membrane following illumination

Membrane potential changes following illumination of a photoreceptor cell in the lateral ocellus of a barnacle (Balanus eburneus) were studied by means of intracellular recording and polarization techniques. Illumination produces a depolarizing response. When the illumination is terminated, the membrane potential temporarily becomes more negative than the resting potential prior to illumination. Although the amplitude of this postillumination hyperpolarization depends upon the intensity as well as the duration of the light pulse, the time course is fairly constant. The hyperpolarization is not associated with any significant membrane conductance increase and is abolished by 10^-5 M ouabain. It diminishes when the external Na or K ions are removed. An intracellular injection of Na ions produces a hyperpolarization similar to that following illumination. It is suggested that the postillumination hyperpolarization is produced by an electrogenic Na pump which is activated by the Na influx during illumination.     

Light-Induced Changes in Photoreceptor Membrane Resistance and Potential in Gecko Retinas : II. Preparations with Active Lateral Interactions

The time-course of light-induced changes in membrane voltage and resistance were measured in single photoreceptors in eyecup preparations of Gekko gekko. A small circular stimulus directed toward the impaled receptor produced membrane hyperpolarization. Application of a steady annular light to the receptor periphery resulted in diminution of the receptor's response to the stimulus. The effects of illumination of the surrounding receptors were isolated by directing a small, steady desensitizing light to the impaled receptor and then applying a peripheral stimulus. Brief stimuli produced a transient decrease in resistance with rapid onset and offset, a time-course similar to that of the response diminution. For some cells a depolarization that coincided with the resistance decrease was seen. During illumination with prolonged stimuli the resistance decrease was followed by a slow increase. After offset resistance rose transiently above the original value and then returned slowly to its original value. The slow resistance changes were not accompanied by changes in membrane voltage. The response diminution, resistance decrease, and depolarization were not observed in retinas treated with aspartate or hypoxia. It is therefore concluded that these effects are mediated by horizontal cells. The diminution is achieved by shunting the receptor potential and may play a role in field adaptation.     

Membrane potential, action potential and activation potential of eggs of the sea urchin, Lytechinus variegatus

Unfertilized Lytechinus variegatus eggs in sea water in their normal physiological state have membrane potentials that approximate ?70 to ?80 mV. This conclusion is based on microelectrode measurements and on computation from the Na⁺ and K⁺ fluxes. The ?8 to ?15 mV values for the membrane potential previously reported and which are generally measured are the consequence of depolarization by impalement. The activation potential in inseminated eggs with an initial membrane potential more negative than ?60 mV is a compound event involving sperm-induced as well as voltage dependent conductance changes. The sperm-induced mechanism is a two-phase conductance increase which involves both Na⁺ and Ca²⁺ during the first phase, and Na⁺ alone during the second phase. In addition, the sperm-induced depolarization at the beginning of the first phase activates a voltage dependent Ca²⁺-conductance mechanism resulting in generation of an action potential.     

Biphasic potassium contractures in frog muscle fibers

Potassium-induced contractures were studied in single fibers from the semitendinosus muscle of Rana pipiens. Contractures elicited by solutions containing 60–117 mM potassium and 120 mM chloride were biphasic, consisting of a rapid initial contraction with a duration at 23°C of less than 1 sec followed by a slow response with a duration of many seconds. At 13°C, the initial response was greatly prolonged so that the two responses virtually fused into a single smooth contracture. Membrane potential in high potassium, high chloride solutions underwent a transient peak depolarization, probably as a result of time-dependent changes in membrane conductance during depolarization. It is proposed that this complex time course of depolarization gives rise to the biphasic contracture response.     

Changes in membrane characteristics of heart muscle during inhibition

Membrane characteristics were studied in isolated muscle strands from auricles of frogs using the "square pulse" technique. Changes in the time course and spatial spread of subthreshold electrotonic potentials were measured. If acetylcholine is applied in concentrations which cause slowing or stoppage of the heart beat, the following changes are produced: (a) the length constant (λ) of the membrane is reduced, (b) the time constant is shortened. The effects are reversible and increase with acetylcholine concentration. The membrane changes caused by acetylcholine dimmish with time. It is concluded that during acetylcholine inhibition, as well as during vagal inhibition, the conductance of the muscle membrane is increased. Appreciable changes in the resting membrane potential need not accompany inhibition.     

Adaptation in the Ventral Eye of Limulus is Functionally Independent of the Photochemical Cycle, Membrane Potential, and Membrane Resistance

The early receptor potential (ERP), membrane potential, membrane resistance, and sensitivity were measured during light and/or dark adaptation in the ventral eye of Limulus. After a bright flash, the ERP amplitude recovered with a time constant of 100 ms, whereas the sensitivity recovered with an initial time constant of 20 s. When a strong adapting light was turned off, the recovery of membrane potential and of membrane resistance had time-courses similar to each other, and both recovered more rapidly than the sensitivity. The receptor depolarization was compared during dark adaptation after strong illumination and during light adaptation with weaker illumination; at equal sensitivities the cell was more depolarized during light adaptation than during dark adaptation. Finally, the waveforms of responses to flashes were compared during dark adaptation after strong illumination and during light adaptation with weaker illumination. At equal sensitivities (equal amplitude responses for identical flashes), the responses during light adaptation had faster time-courses than the responses during dark adaptation. Thus neither the photochemical cycle nor the membrane potential nor the membrane resistance is related to sensitivity changes during dark adaptation in the photoreceptors of the ventral eye. By elimination, these results imply that there are (unknown) intermediate process(es) responsible for adaptation interposed between the photochemical cycle and the electrical properties of the photoreceptor.     

Reduction of anion reversal potential subverts the inhibitory control of firing rate in spinal lamina I neurons: towards a biophysical basis for neuropathic pain

Background

Reduction of the transmembrane chloride gradient in spinal lamina I neurons contributes to the cellular hyperexcitability producing allodynia and hyperalgesia after peripheral nerve injury. The resultant decrease in anion reversal potential (i.e. shift in E _anion to less negative potentials) reduces glycine/GABA_A receptor-mediated hyperpolarization, but the large increase in membrane conductance caused by inhibitory input can nonetheless shunt concurrent excitatory input. Without knowing the relative contribution of hyperpolarization and shunting to inhibition's modulation of firing rate, it is difficult to predict how much net disinhibition results from reduction of E _anion. We therefore used a biophysically accurate lamina I neuron model to investigate quantitatively how changes in E _anion affect firing rate modulation.

Results

Simulations reveal that even a small reduction of E _anion compromises inhibitory control of firing rate because reduction of E _anion not only decreases glycine/GABA_A receptor-mediated hyperpolarization, but can also indirectly compromise the capacity of shunting to reduce spiking. The latter effect occurs because shunting-mediated modulation of firing rate depends on a competition between two biophysical phenomena: shunting reduces depolarization, which translates into reduced spiking, but shunting also shortens the membrane time constant, which translates into faster membrane charging and increased spiking; the latter effect predominates when average depolarization is suprathreshold. Disinhibition therefore occurs as both hyperpolarization- and shunting-mediated modulation of firing rate are subverted by reduction of E _anion. Small reductions may be compensated for by increased glycine/GABA_A receptor-mediated input, but the system decompensates (i.e. compensation fails) as reduction of E _anion exceeds a critical value. Hyperexcitability necessarily develops once disinhibition becomes incompensable. Furthermore, compensation by increased glycine/GABA_A receptor-mediated input introduces instability into the system, rendering it increasingly prone to abrupt decompensation and even paradoxical excitation.

Conclusion

Reduction of E _anion dramatically compromises the inhibitory control of firing rate and, if compensation fails, is likely to contribute to the allodynia and hyperalgesia associated with neuropathic pain. These data help explain the relative intractability of neuropathic pain and illustrate how it is important to choose therapies not only based on disease mechanism, but based on quantitative understanding of that mechanism.     

Interpretation of the Repetitive Firing of Nerve Cells

Eccentric cells of Limulus respond with repetitive firing to sustained depolarizing currents. Following stimulation with a step of current, latency is shorter than first interval and later intervals increase progressively. A shock of intensity twice threshold can evoke firing 25 msec. after an impulse. But in the same cell, a current step twice rheobase evokes a second impulse more than 50 msec. after the first, and current intensity must be raised to over five times rheobase to obtain a first interval of about 25 msec. Repetitive firing was evoked by means of trains of shocks. With stimuli of moderate intensity, firing was evoked by only some of the shocks and intervals between successive impulses increased with time. This is ascribed to accumulation of refractoriness with successive impulses. Higher frequencies of firing are obtained with shocks of intensity n x threshold than with constant currents of intensity n x rheobase. It is concluded that prolonged currents depress the processes leading to excitation and that (in the cells studied) repetitive firing is controlled both by the after-effects of firing (refractoriness) and by the depressant effects of sustained stimuli (accommodation). Development of subthreshold "graded activity" is an important process leading to excitation of eccentric cells, but is not the principal factor determining frequency of firing in response to constant currents.     

Conditions of appearance of local response and action potential in a simple mathematical model of nerve

A simple mathematical model of a nerve fibre is proposed. According to the model the current-voltage relation for the peak Na-current is represented as a broken line. It is shown that within the potential range, where the differential (slope) Na-conductance isnegative but its absolutevalue is smaller than the membrane conductance, the fibre gives a subthreshold local response. Within the range, where differential Na-conductance is negative and has absolute value larger than the membrane conductance, the action potential arises, the rise being independent on the strength of the stimulus and conditioned only by the membrane parameters. A simple formula is obtained which describes the relation between the strength of the threshold stimulus, I, and its duration, t₀. There are two parameters in this formula, which can be measured independently: the time constant of the membrane and the lag-period in the Na-current. Insertion of these parameters into the formula allows one to construct a theoretical I-t₀ curve in good agreement with experiment.     

Membrane and firing properties of avian medial vestibular nucleus neuronsin vitro

The intrinsic membrane and firing properties of medial vestibular nucleus (MVN) neurons were investigated in slices of the chick brainstem using intracellular recording and current injection. Avian MVN neurons fired spontaneous action potentials with very regular interspike intervals. The rapid repolarization of all action potentials was followed by an after-hyperpolarization. Intracellular injection of steps of hyperpolarizing current revealed both an inward rectification of the membrane potential during the step and a rebound depolarization following the offset of the step. In some neurons, the rebound depolarization resulted in bursts of action potentials. Steps of depolarizing current applied to spontaneously active neurons evoked increases in firing rate that were higher at the onset of the step than during the steady-state response. The relationship between current and firing rate was linear. The membrane and firing properties of avian MVN neurons were distributed continuously across the population of recorded neurons. These properties appear identical to those of rodent MVN neurons, suggesting that the composition and distribution of ion channels in the MVN neuronal membrane has been highly conserved across vertebrate species.Abbreviations MVN medial vestibular nucleus - VOR vestibulo-ocular reflex - AHP after-hyperpolarization     

K Permeability of Nitella clavata in the Depolarized State

Membrane current responses to sudden potential changes were recorded in solutions of various [K]_o on 52 internodal cells of Nitella clavata. The membrane current after sudden depolarization had a component sensitive to [K]_o which increased with time from 0.3 to 2.0 s and remained steady thereafter. This late current became zero at values of E and [K]_o which suggests that the current was nearly all carried by K⁺. The potassium conductivity represented by this current increased with depolarization, with a half-maximum value at about -70 mV, and saturation at about -30 to -20 mV. The potassium conductance also increased with increasing [K]_o, but less rapidly than predicted for constant potassium permeability. This failure of the conductance to increase with [K]_o was relatively the same at all membrane potentials and may be explained by a model with a finite number of channels. No attempt was made to model the dependence of g_K on time after depolarization or on membrane potential. However, the finding that the membrane potential did not affect the way in which the permeability depended on [K]_o suggests that the membrane potential change does not affect the affinity of the sites, and that the increase in g_K with time after depolarization is brought about by an increase in the number of channels with such sites.     

Modulatory action of acetylcholine on the Na-dependent action potentials in Kenyon cells isolated from the mushroom body of the cricket brain

Kenyon cells, intrinsic neurons of the insect mushroom body, have been assumed to be a site of conditioning stimulus (CS) and unconditioned stimulus (US) association in olfactory learning and memory. Acetylcholine (ACh) has been implicated to be a neurotransmitter mediating CS reception in Kenyon cells, causing rapid membrane depolarization via nicotinic ACh receptors. However, the long-term effects of ACh on the membrane excitability of Kenyon cells are not fully understood. In this study, we examined the effects of ACh on Na⁺ dependent action potentials (Na⁺ spikes) elicited by depolarizing current injection and on net membrane currents under the voltage clamp condition in Kenyon cells isolated from the mushroom body of the cricket Gryllus bimaculatus. Current-clamp studies using amphotericin B perforated-patch recordings showed that freshly dispersed cricket Kenyon cells could produce repetitive Na⁺ spikes in response to prolonged depolarizing current injection. Bath application of ACh increased both the instantaneous frequency and the amplitudes of Na⁺ spikes. This excitatory action of ACh on Kenyon cells is attenuated by the pre-treatment of the cells with the muscarinic receptor antagonists, atropine and scopolamine, but not by the nicotinic receptor antagonist mecamylamine. Voltage-clamp studies further showed that bath application of ACh caused an increase in net inward currents that are sensitive to TTX, whereas outward currents were decreased by this treatment. These results indicate that in order to mediate CS, ACh may modulate the firing properties of Na⁺ spikes of Kenyon cells through muscarinic receptor activation, thus increasing Na conductance and decreasing K conductance.     

Electrophysiological study of the action mechanism of somatoliberin (GH-RH) on hypophyseal GH3 tumor cells]

We have investigated the electrical response of patched GH3 cells to Growth-Hormone Releasing-Hormone (GH-RH). GH-RH (100 nM) enhanced firing frequency of action potentials. This is accompanied by membrane depolarization (5-10 mV) and conductance increase. Voltage clamp studies reveal that GH-RH potentiates calcium inward currents and a calcium-dependent chloride current; transient outward current is diminished. These changes in membrane conductance account for the cytosolic free calcium rise shown by Indo-1 fluorescence measurements.     

A study of the crustacean axon repetitive response. 3. A comparison of the effect of veratrine sulfate solution and potassium-rich solutions

The crustacean single nerve fiber gives rise to trains of impulses during a prolonged depolarizing stimulus. It is well known that the alkaloid veratrine itself causes a prolonged depolarization; and consequently it was of interest to investigate the effect of this chemically produced depolarization on repetitive firing in the single axon and compare it with the effect of depolarization by an applied stimulating current or by a potassium-rich solution. It was found that veratrine depolarization, though similar in some respects to a potassium-rich depolarization of depolarizing current effect, was in many respects quite different. (1) At low veratrine concentration, less than 1 Mg%, the negative after potential following a spike action potential was prolonged and augmented. At higher concentrations or after a long period of time, veratrine caused a prolonged steady state depolarization of the membrane, the “veratrine response”. The prolonged plateau depolarization response could be elicited with or without an action potential spike by a short or long duration stimulating pulse, but only if the veratrine depolarization was prevented or offset by an applied conditioning hyperpolarizing inward current. (2) The “veratrine response” resembled the potassium-rich solution response in the plateau-like contour of the depolarization and the very low membrane resistance during this plateau phase. Like the potassium response, it was possible to obtain a typical hyperpolarizing response with an inwardly directed current pulse if applied during the plateau phase. During the negative after potential augmented with veratrine, however, this hyperpolarizing response was not observed. (3) In contrast to the potassium response, however, the “veratrine response” is intimately associated with the sodium concentration in the external medium. The depolarization in millivolts is linearly related to the log of the concentration of external sodium. Moreover, during veratrine action there is a continuous and progressive inactivation of the sodium mechanism which ultimately terminates repetitive firing and abolishes the spike action potential. Then even with conditioning hyperpolarization only the slow response may be elicited in veratrine, occasionally with a spike superimposed if sodium is present, but without repetitive firing. (4) It is concluded that veratrine action is the result of a chemical or metabolic reaction by the alkaloid in the membrane. It is suggested that veratrine may inhibit the sodium extrusion mechanism, or may itself compete for sites in the membrane with calcium and/or sodium. This explains the inhibiting effect of high calcium, the abolition of the “veratrine response” with low temperature and high calcium combined and the progressive inactivation of the sodium system.     

Electromotive force of Nitella membrane during excitation

Excitation of the Nitella membrane is analysed by assuming themembrane to be an electromotive force in series with a resistance,both being variables of time and of membrane potential. Duringstep depolarization beyond a threshold, conductance and electromotiveforce increase transiently, finally reaching their respectivesteady state levels. The conductance increase peak is attainedearlier than the peak for electromotive force increase. Wheneverelectromotive force increases beyond the level of clamped membranepotential, the ionic current flows inward. This is consideredto be the origin of the apparent negative resistance characteristicof the excitable membrane. Anodal break response and spontaneousfiring of Nitella membrane are also caused by transient increasesin electromotive force and conductance irrespective of whetherthe membrane potential is being held at its resting level. Thetransient increase in electromotive force reflects changes,like a phase transition, occurring during excitation. (Received May 6, 1968; )     

Diversity of Intrinsic Frequency Encoding Patterns in Rat Cortical Neurons—Mechanisms and Possible Functions

Extracellular recordings of single neurons in primary and secondary somatosensory cortices of monkeys in vivo have shown that their firing rate can increase, decrease, or remain constant in different cells, as the external stimulus frequency increases. We observed similar intrinsic firing patterns (increasing, decreasing or constant) in rat somatosensory cortex in vitro, when stimulated with oscillatory input using conductance injection (dynamic clamp). The underlying mechanism of this observation is not obvious, and presents a challenge for mathematical modelling. We propose a simple principle for describing this phenomenon using a leaky integrate-and-fire model with sinusoidal input, an intrinsic oscillation and Poisson noise. Additional enhancement of the gain of encoding could be achieved by local network connections amongst diverse intrinsic response patterns. Our work sheds light on the possible cellular and network mechanisms underlying these opposing neuronal responses, which serve to enhance signal detection.     

Two Light-Induced Processes in the Photoreceptor Cells of Limulus Ventral Eye

The dark-adapted current-voltage (I-V) curve of a ventral photoreceptor cell of Limulus, measured by a voltage-clamp technique, has a high slope-resistance region more negative than resting voltage, a lower slope-resistance region between resting voltage and zero, and a negative slope-resistance region more positive than 0 v. With illumination, we find no unique voltage at which there is no light-induced current. At the termination of illumination, the I-V curve changes quickly, then recovers very slowly to a dark-adapted configuration. The voltage-clamp currents during and after illumination can be interpreted to arise from two separate processes. One process (fast) changes quickly with change in illumination, has a reversal potential at +20 mv, and has an I-V curve with positive slope resistance at all voltages. These properties are consistent with a light-induced change in membrane conductance to sodium ions. The other process (slow) changes slowly with changes in illumination, generates light-activated current at +20 mv, and has an I-V curve with a large region of negative slope resistance. The mechanism of this process cannot as yet be identified.     

Decreased Response to Acetylcholine during Aging of Aplysia Neuron R15

How aging affects the communication between neurons is poorly understood. To address this question, we have studied the electrophysiological properties of identified neuron R15 of the marine mollusk Aplysia californica. R15 is a bursting neuron in the abdominal ganglia of the central nervous system and is implicated in reproduction, water balance, and heart function. Exposure to acetylcholine (ACh) causes an increase in R15 burst firing. Whole-cell recordings of R15 in the intact ganglia dissected from mature and old Aplysia showed specific changes in burst firing and properties of action potentials induced by ACh. We found that while there were no significant changes in resting membrane potential and latency in response to ACh, the burst number and burst duration is altered during aging. The action potential waveform analysis showed that unlike mature neurons, the duration of depolarization and the repolarization amplitude and duration did not change in old neurons in response to ACh. Furthermore, single neuron quantitative analysis of acetylcholine receptors (AChRs) suggested alteration of expression of specific AChRs in R15 neurons during aging. These results suggest a defect in cholinergic transmission during aging of the R15 neuron.     

Generator processes of repetitive activity in a pacinian corpuscle

Response patterns resulting from repetitive mechanical stimulation of the corpuscle depend on (1) the time course of recovery of the generator potential, on (2) the recovery of critical firing height, and on (3) the stimulus strength/generator potential function. By either augmenting stimulus frequency at constant strength, or by reducing strength at constant frequency, a sequence of propagated potentials is turned into a pattern of alternating regenerative and generator responses. In such a pattern an extra impulse can be set up whenever an extra stimulus produces a generator potential of enough amplitude to reach the firing height of the corresponding period. The new requirements of firing height introduced by the refractory trail of the extra impulse determine resetting of periodicity and appearance of a "compensatory pause." The decay time of the single generator potential is independent of stimulus duration. This is interpreted as a factor determining receptor adaptation. Upon repetitive stimulation at intervals above ½ decay time of the single generator potential, a compound generator potential is built up which shows no spontaneous decline. However, in spite of being considerably greater than the firing height for single impulses, the constant level of depolarization of the compound generator potential is unable to produce propagated potentials. A hypothesis is brought forward which considers the generator potential to arise from membrane units with fluctuating excitability scattered over the non-myelinated nerve ending.