Electrical phenomena in nerve; crab nerve

The resting and action potentials of the leg nerves of the spider crab are reduced by procaine, cocaine, iodoacetate, KCl, and veratrine. The first three agents depress the sensitivity of the resting potential to anoxia, while the last can be shown to augment it. Glucose sustains activity and the polarized state in the absence of oxygen, an effect blocked by iodoacetate; corresponding concentrations of lactate and pyruvate are inert under most experimental conditions. DDT and veratrine both induce repetitive activity following an impulse, but only the latter does so with a marked increase in negative after-potential. The negative after-potential induced by veratrine is decreased by KCl relatively more than the spike or the resting potential. Elevation of the calcium content of the medium increases this after-potential. Neither ion appreciably alters the time constant of repolarization. The recovery is more rapid than that obtained following prolonged activity of both veratrinized and unveratrinized nerves. Repolarization following a tetanus is accelerated by an increase in the volume of solution in contact with the fibers; associated with this is an augmentation of the positive after-potential which normally follows a bout of activity. Yohimbine induces a positive after-potential following individual spikes which is depressed by an elevation of the potassium or calcium content of the medium. These observations are discussed from the standpoint of the available evidence for the involvement of potassium at the surface of the fibers as regulated by a labile permeability and metabolism. The potassium liberated by the action potential, calculated from the polarization changes, agrees closely with an available analytical figure; less direct observations are also found to be consistent with this view.     

Properties of action potentials in Drosera tentacles

Summary Action potentials of Drosera tentacles resemble those of vertebrate peripheral nerves in that they appear to be comprised of relatively uniform spikes, variable shoulders or negative after-potentials, and variable positive after-potentials. The peaking of the spike corresponds to a period of great refractoriness, while action potentials of low amplitude may be fired readily during the negative after-potential. The action potentials fired during the negative after-potential appear to be unlike those of peripheral nerves in that they are of abnormally brief duration. Also apparently different from the case in peripheral nerves is the dependence of the duration of an action potential on the interval separating it from the preceding action potential.Action potentials propagate from the neck of the stalk to its base at about 5 mm s^-1 at room temperature. Propagation may be reversed artificially, consistent with the possibility that the neuroid cells are electrically coupled.     

Low level impedance changes following the spike in the squid giant axon before and after treatment with "veratrine" alkaloids

The increase in conductance, which accompanies the spike in the presence of sea water, is followed by a decrease to below the resting level, here designated as the "initial after-impedance," which lasts 3 msec. and is 3 per cent as great as the increase. Treatment with cevadine usually obliterates the latter but leaves the former essentially unaltered. In addition, the alkaloid gives rise to periodic conductance increases followed by a prolonged, exponentially decaying elevated conductance (the "negativity after-impedance") which correspond closely to potential oscillations and to the negative after-potential. These are also only a few per cent of the major conductance change. Veratridine causes a conductance increase which lasts longer and which also conforms closely with earlier after-potential results. Preliminary calculations indicate that the negativity after-impedance and the negative after-potential may be due to the subsidence of an elevated chloride permeability. However, no satisfactory explanation is available for the initial after-impedance or for the temporal course of the conductance changes associated with oscillations in membrane potential.     

After-currents,after-potentials,excitability, and ventral root electrotonus in spinal motoneurons

The spinal cord constitutes a volume conductor. Potential changes are recorded therefrom only as current flows. During the period of the after-potentials current flows in significant density only if the after-polarization differs at different points of the active neurons. Thus one does not record after-potentials in volume; one may record after-currents which are defined as the resultants of differences in after-potentials. Measurable excitability change during the period of the after-potentials, in the event no current flows, might be regarded as approximating the change of intrinsic polarization status at the region tested. In the presence of after-current flow excitability change would approximate the sum of intrinsic change and extrinsic change due to current flow. In giving rise by differences to current flow after-potentials come to act as agents, and events in one part of a neuron help to determine excitability in other parts. Since the intramedullary after-current flow is not the after-potential of the soma, it follows that ventral root electrotonus which results from axonal after-current flow cannot be considered the counterpart of somatic after-potential. Following conduction of an antidromic volley after-current flows between somata and axons. According to the signs of the recorded potential changes, after-current flow initially, and for approximately 45 msec., is in the direction from somata to axons. Thereafter, and for approximately another 75 msec., the direction of flow is reversed. During the period of after-current flow following antidromic conduction the excitability of neighboring motoneurons is altered in a manner that reproduces the phases of after-current flow. The initial phase, depression, was first described by Renshaw. The after-potentials of ventral root fibers have been studied. In a single action and in usual form, they consist of a negative after-potential of considerable magnitude and of some 35 msec. duration, and a positive after-potential detectable for approximately 120 msec. Variants and the influence of temperature change are described. The recovery cycle of ventral root axons in general compares with the after-potential cycle. Recovery of intramedullary motor axons differs from that of their extramedullary projections as ventral root fibers in a manner that is accountable to intramedullary flow of after-current. Since the intrinsic recovery process of the motoneuron somata cannot be measured in the presence of current flow it must be estimated by correcting the observed recovery for the influence of known current flows. When this is done the resultant in simplest form provides for intrinsic somatic recovery from refractoriness through a single phase of subnormality lasting some 60 msec. Conditions for the relatively undistorted recording of antidromic ventral root electrotonus are described. They include provisions that the proximal ventral root electrode must be within 12 mm. of the root-cord junction and that the distal electrode must be located in excess of 30 mm. from the distal severed end of the ventral root. Antidromic ventral root electrotonus is a counterpart of the current flows in the intramedullary stretch of the axons. Initially, during the phase of metadromal postivity of the intramedullary axons, electrotonus is negative. During the period of deflections Sp-An, that signify after-current flow into the axons, electrotonus is positive. Finally during the period of deflections Sn-Ap, that signify after-current flow outwards through the intramedullary axon membranes, electrotonus is negative. Electrotonic showing is not of sufficient magnitude to make the time course of ventral root electrotonus palpably different from that of the generating intramedullary currents.     

Oscillatory zones and their role in normal and abnormal sheep purkinje fiber automaticity

The mechanisms by which low [K(+)](o) induces spontaneous activity was studied in sheep Purkinje fibers. Purkinje strands were superfused in vitro and membrane potentials were recorded by means of a microelectrode technique. The results show that low [K(+)](o) increases the slope and amplitude of early diastolic depolarization, sharpens the transition between early and late diastolic depolarizations, induces an after-potential and large pre-potentials through a negative shift of an oscillatory zone. Pre-potentials occur progressively sooner during diastole and merge with the after-potential to induce uninterrupted spontaneous discharge. During recovery, when the rate slows, after- and pre-potentials separate once more, the slower discharge decreasing the after-potentials but not the pre-potentials. Low [K(+)](o) has little effect on the plateau, but markedly slows phase 3 repolarization and may altogether prevent it. At depolarized levels, voltage oscillations, slow responses, sinusoidal fluctuations or quiescence may be present depending on voltage. During the recovery, a train of either sub-threshold oscillations or spontaneous action potentials appear towards the end of phase 3 repolarization. The cessation of the action potentials unmasks large sub-threshold oscillations, that occur in the oscillatory zone. Drive, high [Ca(2+)](o) and norepinephrine increase slope and amplitude of early diastolic depolarization as low [K(+)](o) does. In low [K(+)](o), Cs(+) prevents spontaneous discharge at polarized levels, but not the decrease in resting potential nor the onset of slow responses at depolarized levels. Cs(+) blocks the train of oscillations and of action potentials occurring during recovery. We conclude that low [K(+)](o) steepens early diastolic depolarization and increases its amplitude through an after-potential that results from an increased Ca(2+) load; allows the attainment of the threshold through Cs(+)-sensitive voltage oscillations which develop when the oscillatory zone is entered either by diastolic depolarization or by phase 3 repolarization; and causes voltage oscillations also at depolarized levels, but through a Cs(+)-insensitive different mechanism.     

Potassium movement in relation to nerve activity

The depolarization of crab nerve during repetitive stimulation is unaffected by the presence of glucose or by an increase in the calcium content of the medium. It is increased in both amplitude and rate by veratrine; in the presence of this alkaloid mixture the rate but not the magnitude of the depolarization is increased by an elevation in the calcium concentration. Repolarization following stimulation is unaltered by glucose and accelerated by a greater calcium concentration. Veratrine increases both the amplitude and the time constant of repolarization; its effect on the time constant is counteracted by an elevation of the calcium in the medium. Potassium released during stimulation and its reabsorption following activity have been observed by analyses of small volumes of sea water in contact with crab nerve. Under the conditions employed 3 x 10(-8) microM potassium is liberated per impulse per gm. wet weight of nerve. This loss is increased by low concentrations of veratrine, which also increase the amount reabsorbed during recovery. The depletion of potassium from the medium is appreciably less if the potassium previously released during activity has not been removed. Inexcitability resulting from anoxia can be washed away with oxygen-free solution-rapidly and completely in the case of the squid axon, slowly and incompletely in crab nerve. The potassium shifts are in the proper direction and of the correct order of magnitude to account for the negative and positive after-potentials in terms of potassium accumulation or depletion in the extracellular space.     

Calcium oscillations: phenomena, mechanisms and significance

Many hormones that mobilise intracellular calcium via inositol 1,4,5 trisphosphate induce oscillations in cytoplasmic free Ca. Two basic oscillatory patterns occur: quasi-sinusoidal oscillations and repetitive free Ca transients. The mechanisms responsible for generating these oscillations are not clear; calcium-induced calcium release, interplay between two intracellular calcium pools and repetitive generation of InsP3 are discussed. The significance of different oscillatory patterns induced by different agonists in the same cell is emphasised, and mechanisms by which the oscillators may retain-receptor specific information are proposed, such as negative feedback onto receptors or G-proteins by protein kinase C. Reasons why cells generate free Ca oscillations and possible consequences such as oscillations in downstream pathways are explored. The possibility that pathological conditions such as aluminium toxicity are exerted through distortion of oscillatory free Ca signalling is raised.     

AHP's, HAP's and DAP's: How Potassium Currents Regulate the Excitability of Rat Supraoptic Neurones

We have constructed mathematical models of the electrical activity of two hypothalamic supraoptic neuro-secretory cell-types, and we support our models with new calcium imaging and in vitro electrophysiological data. These cells are neurones that project to the pituitary gland and secrete either of two hormones, oxytocin or vasopressin, into the blood from their axonal terminals. Oxytocin-secreting and vasopressin-secreting cells are closely related and physically they differ only subtly, however when physiologically stressed their discharge patterns are dramatically distinct. We first show how each potassium current contributes to the action-potentials and after-potentials observed in these cells, and we show how these after-potentials are correlated to intra-cellular calcium elevations. We then show how these currents regulate the excitability of these cells and consequently shape their discharge pattern.     

Modelling receptor-controlled intracellular calcium oscillators.

This paper presents mathematical models for the hepatocyte calcium oscillator which follow the concepts in a class of informal models developed to account for the striking dependence on the receptor type of several features of the calcium oscillations, in particular the shape and duration of the free calcium transients. The essence of these models is that the transients should be timed by a build-up of activated GTP-binding proteins, which, combined with positive feedback processes and perhaps with cooperative effects, leads to a sudden activation of phospholipase C (PLC), followed by negative feedback processes which switch off the calcium rise and lead to a fall in free calcium back to resting levels. These models predict pulsatile oscillations in inositol (1,4,5)P3 as well as in free calcium. We show that receptor-controlled intracellular calcium oscillators involving an unknown positive feedback pathway onto PLC and negative feedback from protein kinase C (PKC) onto G-proteins and receptors, or negative feedback by stimulation of GTPase activity can simulate many of the features of observed intracellular calcium oscillations. These oscillators exhibit a dependence of frequency on agonist concentration and a dependence of transient duration on receptor and G-protein type. We also show that a PLC-dependent GTPase activating factor (GAF) could provide explanations for some otherwise puzzling features of intracellular calcium oscillations.     

The After-Potential that Follows Trains of Impulses in Frog Muscle Fibers

Intracellular recordings of the negative after-potential that follows a train of impulses in frog muscle are described. It is postulated that the cause of this after-potential is an accumulation of potassium, during the train, in an intermediary space that is located between the major portion of the sarcoplasm and the external fluid. The decline of the after-potential is then produced by the exchange of potassium from the intermediary space with sodium from the external fluid.     

The inhibitory effects of dantrolene on action potential-induced calcium transients in cultured rat dorsal root ganglion neurons

We investigated the actions of dantrolene Ca(2+)-induced on Ca(2+)-release (CICR) evoked by action potentials in cultured rat sensory neurons. The effect of dantrolene on action potential after-depolarization and voltage-activated calcium currents was studied in cultured neonatal rat dorsal root ganglion cells (DRG) using the whole-cell patch-clamp technique. Depolarizing current injection evoked action potentials and depolarizing after-potentials, which are activated as a result of CICR following a single action potential in some cells. The type of after-potentials was determined by inducing action potentials from the resting membrane potential. Extracellular application of dantrolene (10 microM) abolished after-depolarizations without affecting action potential properties. Furthermore, dantrolene significantly reduced repetitive action potentials after depolarizing current injection into these neurons, but had no significant effect on the steady-state current voltage relationship of calcium currents in these neurons. We conclude that dantrolene inhibits the induction of action potential after depolarizations by inhibiting CICR in cultured rat sensory neurons.     

Classification of vestibulo-spinal neurons of the cat Deiters' nucleus

Antidromic excitation of neurons of the lateral vestibular nucleus of Deiters in cats in response to stimulation of the vestibulo-spinal tract in the cervical segments of the spinal cord was studied by intracellular microelectrode recording. Individual components of the antidromic action potential and accompanying after-potentials were analyzed and fast and slow neurons distinguished. The vestibulo-spinal neurons were differentiated on the basis of after-potentials accompanying the antidromic action potential. The ratio between fast and slow neurons differed in individual groups. The parameters of the depolarization after-potentials were directly proportional to the duration of the refractory period of the neurons studied. An attempt was made to correlate differences in the responsiveness of neurons with an identical conduction velocity along their axons with the characteristics of the depolarization after-potential.     

The action potential of spinal axons in vitro

Despite the trauma of dissection and special metabolic requirements, the physiological properties of funiculi of the mammalian spinal cord can be studied in vitro. They are adequately oxygenated by diffusion at 0.88 atm. pO(2) and remain in a functionally normal state for over 12 hours. The internal consistency of several kinds of data presented in this and the foregoing papers (5, 38) serves to characterize certain properties of central myelinated axons whether excised or in situ. (1) Spinal tracts support a large spike potential in vitro whose form, duration, and velocity are comparable to those of alpha fibers in vitro and spinal tracts in vivo. (2) Properties consistent with a large L fraction are found in central axons whether excised or in situ. (3) Following conduction there has been identified post-spike supernormality with exponential time course (7.5 msecs. half-time) which is the result of activity intrinsic to parent fibers of dorsal columns. The supernormality is similar in form and magnitude both in excised and intact funiculi. (4) In excised funiculi the action potential of parent axons includes a large negative after-potential whose form and duration correspond satisfactorily with this supernormality. This potential appears not to result from activity arising in broken collaterals. (5) Central axons, excised or intact, fire spontaneously in the presence of citrate ion, and when synchronized by stimulation develop periodic oscillations at about 400 C.P.S. but show no such behavior in the presence of excess potassium ion. Certain characteristics peculiar to central axons indicate that they occupy an extreme position in the spectrum of properties encountered in conducting tissues. Dorsal column myelinated axons differ from their peripheral counterparts, even though they are parts of the same cell, in the following ways. The maintenance of the column spike potential is more critically dependent on CO(2) and the entire tissue mass has a higher oxygen consumption. The negative after-potential is much larger and the positive after-potential, non-existent following a single volley, is more difficult to develop by repetitive stimulation. Unlike peripheral nerve, central axons are not incited to spontaneous activity by manipulation of certain constituents normally present in their environment. However, when induced by the application of citrate the resulting rhythmic behavior has twice the frequency of that in peripheral nerve. In general, the recovery process in central axons is more invariant than that in peripheral axons when they are subjected to similar changes in their artificial environments.     

Further studies on the functional properties of spinal axons in vivo

Mammalian spinal tracts in situ demonstrate a phase of marked hyperexcitability during hypoxia or on the application of an excess of potassium or citrate ion. This is in keeping with the fact that they also show post-spike supernormality as well as hyperexcitability under cathodal polarization (17). Behavior of this kind indicates that central axons carry a well developed L fraction of membrane properties. The rhythmic state in central axons in situ, unlike peripheral nerve or spinal root, is not induced by the action of excess potassium ion. This appears to be related to the absence of a positive after-potential in dorsal columns (17). However, sodium citrate can elicit autonomous firing in central axons. When synchronized by an applied stimulus the resulting periodic oscillations have a fundamental frequency (340 to 400 C.P.S.) which is significantly greater than that of peripheral nerve.     

Microwave enhancement of membrane conductance: effects of EDTA, caffeine and tetracaine

Effects of tetracaine and caffeine on snail neurons were studied. They displayed depolarization and an increase of membrane conductance. In addition, tetracaine diminished membrane time constant whereas caffeine augmented hyperpolarizing after-potential. It was also shown that tetracaine blocks the caffeine effect. Microwave irradiation of snail neurons enhanced membrane conductance. This effect was not observed in neurons treated with tetracaine or injected with EDTA. Analysis of these results points to intracellular free calcium as a possible trigger of snail neuron microwave response.     

The influence of high hydrostatic pressure on cocaine and veratrine action in a vertebrate nerve

The application of high hydrostatic pressure to toad sciatic nerve causes a gain in sodium and a loss of potassium which are not affected by cocaine. However, cocaine action is enhanced by high pressure when counteracting veratrine depolarization and when blocking the action potential. Various effects of elevated pressure on the after-potentials are presented and the role of ions in these processes is discussed.     

Neighborhood-Scale Analyses of Non-additive Species Effects on Cation Concentrations in Forest Soils

Trees affect soil chemistry and nutrient availability via a broad range of processes. Effects can vary dramatically among species, whose distinctive spatial “footprints” can vary for different nutrients. Potentially overlapping effects of neighboring trees in mixed-species stands make footprint shape and interspecific interactions important: If interactions are non-additive, then not only abundance but also spatial configuration influence tree species’ effects on ecosystem properties. We used spatially explicit neighborhood-scale data on tree distributions to fit maximum likelihood models of exchangeable calcium, magnesium, and potassium in surface soils of four sites in northern hardwood forests in northwestern Connecticut, USA. The models incorporated parent material, site, and tree species or functional group configuration to predict availability of the three cations. Site had a stronger effect than species for all cations (there was no species effect for potassium), even after accounting for variation in parent material. Species’ spatial footprints extended further from the stem for calcium than magnesium, which is consistent with the relative importance of litterfall versus stemflow transfer of these nutrients. The magnitude of species effects on calcium and magnesium varied widely. Functional groups made up of species with positive or negative effects provided parsimonious models for magnesium and calcium, and the best model for calcium included a non-additive, antagonistic effect whose strength varied by site. This non-additive effect suggests that the degree of intermingling of tree species from negative- and positive-effect functional groups may influence stand-level availability of calcium, a key nutrient for forest health in these ecosystems.     

Synaptic mechanisms of spike generation in bursts by neurons of the carp tectum and olfactory bulb

Synaptic mechanisms of burst activity generation in certain neurons of the tectum opticum and mechanisms of generation of stimulation-induced group discharges by certain secondary neurons of the olfactory bulb were analyzed in carp (Cyprinus carpio L.). Spikes of the spontaneous discharge in neurons of the tectum were accompanied by depolarizing after-potentials, which caused the burst discharges of these cells. Evidence is given in support of the synaptic nature of the after-potential; it is suggested that it is generated by a recurrent collateral mechanism. Synaptic bombardment causing the appearance of a group discharge in olfactory bulb neurons and groups of spikes in their spontaneous activity was found to be intermittent in character. These features of unit activity in the olfactory bulb are shown to be connected with the presence of excitatory synaptic interaction between several neurons, probably dendro-dendritic in nature.M. V. Lomonosov Moscow State University. Translated from Neirofiziologiay, Vol. 14, No. 5, pp. 483–490, September–October, 1982.     

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.     

Positive Feedback Promotes Oscillations in Negative Feedback Loops

A simple three-component negative feedback loop is a recurring motif in biochemical oscillators. This motif oscillates as it has the three necessary ingredients for oscillations: a three-step delay, negative feedback, and nonlinearity in the loop. However, to oscillate, this motif under the common Goodwin formulation requires a high degree of cooperativity (a measure of nonlinearity) in the feedback that is biologically “unlikely.” Moreover, this recurring negative feedback motif is commonly observed augmented by positive feedback interactions. Here we show that these positive feedback interactions promote oscillation at lower degrees of cooperativity, and we can thus unify several common kinetic mechanisms that facilitate oscillations, such as self-activation and Michaelis-Menten degradation. The positive feedback loops are most beneficial when acting on the shortest lived component, where they function by balancing the lifetimes of the different components. The benefits of multiple positive feedback interactions are cumulative for a majority of situations considered, when benefits are measured by the reduction in the cooperativity required to oscillate. These positive feedback motifs also allow oscillations with longer periods than that determined by the lifetimes of the components alone. We can therefore conjecture that these positive feedback loops have evolved to facilitate oscillations at lower, kinetically achievable, degrees of cooperativity. Finally, we discuss the implications of our conclusions on the mammalian molecular clock, a system modeled extensively based on the three-component negative feedback loop.