PubMed Health⌕ Search

Biomedical subjects

R Werman

Publications and source records attributed to R Werman.

At least 19 recordsLinked to original sources
PubMed1992

An analysis of the long-lasting after-hyperpolarization of guinea-pig vagal motoneurones.

1. The long-lasting after-hyperpolarization which characterizes the neurones of the dorsal motor nucleus of the vagus in the guinea-pig was studied in vitro. 2. Following a train of action potentials, vagal motoneurones develop a long-lasting after-hyperpolarization. Two different shapes of long-lasting after-hyperpolarization were encountered: an after-hyperpolarization which slowly (0.6-1.2 s) and monotonically developed to peak value; and a second type of long-lasting after-hyperpolarization where the onset of the slow component appears to be masked by an early, relatively fast component. Both shapes of long-lasting after-hyperpolarization depend on Ca2+ influx and increase as a function of the number of action potentials in the train. 3. A novel procedure was used to analyse the ionic processes which underlie the long-lasting after-hyperpolarization. The neuronal responses to a series of long (7 s) hyperpolarizing current pulses during the long-lasting after-hyperpolarization were recorded and the voltage-current curves at 600 different time points along the long-lasting after-hyperpolarization were plotted. The conductance and the reversal potential at each time point were calculated from the slope and the intersection of these curves, respectively. 4. Using this procedure it was found that the long-lasting after-hyperpolarization consists of two conductances that differ in kinetic properties and reversal potential: an early conductance which peaks shortly after the end of the train and decays in a few tenths of seconds (EAHP), and a late conductance which develops slowly (time to peak about 1 s) and decays in 3-8 s (LAHP). The reversal potential for the early conductance is 10 mV more positive than the reversal potential for the late conductance (-84 mV); the latter reversal potential is in agreement with the K+ equilibrium potential. The different shapes of long-lasting after-hyperpolarization can be explained by different ratios of these two conductances. 5. Noradrenaline (10 microM) selectively blocks the late conductance, without an observable effect on the Ca2+ action potential. 6. The behaviour of the noradrenaline-sensitive late conductance was analysed. The amplitude of the conductance change increased sigmoidally as a function of the number of spikes in the train. A log-log plot suggests that at least two Ca2+ ions participate in the opening of a K+ channel. 7. A model that accounts for the slow kinetics of the late conductance was constructed.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗
PubMed1990

Spatial and temporal analysis of calcium-dependent electrical activity in guinea pig Purkinje cell dendrites.

We have used the calcium indicator dye arsenazo III, together with a photodiode array, to record intracellular calcium changes simultaneously from all regions of individual guinea pig cerebellar Purkinje cells in slices. The optical signals, recorded with millisecond time resolution, are good indicators of calcium-dependent electrical events. For many cells the sensitivity of the recordings was high enough to detect signals from each array element without averaging. Consequently, it was possible to use these signals to follow the complex spatial and temporal patterns of plateau and spike potentials. Calcium entry corresponding to action potentials was detected from all parts of the dendritic field including the fine spiny branchlets, demonstrating that calcium action potentials spread over the entire arbor. Usually, the entire dendritic tree fired at once. But sometimes only restricted areas had signals at any one moment with transients detected in different regions at other times. In one cell, six separate zones were distinguished. These results show that calcium action potentials could be regenerative in some dendrites and could fail to propagate into others. Signals from plateau potentials were also detected from extensive areas in the dendritic field but were always smaller than those caused by a burst of action potentials.

Action Potentials↗
PubMed1988

Electrophysiology of degenerating neurones in the vagal motor nucleus of the guinea-pig following axotomy.

1. The electrophysiological properties of motoneurones in the dorsal motor nucleus of the vagus in the guinea-pig were studied at different times following cervical vagotomy. The results were compared both to normal neurones and to results obtained at the same time from intact neurones located in the contralateral nucleus. 2. The input resistances of axotomized neurones are significantly higher than those of normal neurones (66 +/- 29 compared to 45 +/- 17 M omega). This difference was seen during the first month following axotomy without any sign of a time-dependent process. On the other hand, no change in resting potential was observed. 3. Significant reduction in action potential amplitude was observed 1 month after axotomy (from 97.8 +/- 8 to 87 +/- 7 mV) and was followed by slow recovery lasting more than 1 year. Neither the Na+ conductance nor the voltage-dependent K+ conductance responsible for the fast rise and fall of the action potential, respectively, were affected by axotomy. 4. One month after axotomy the action potential duration in axotomized neurones was found to be shorter than that of normal neurones (0.9 +/- 0.1 ms compared to 1.1 +/- 0.04 ms). We show that this decrease in duration reflects a reduction in the depolarizing hump on the falling phase of the action potential, which is known to express the Ca2+ conductance activated during the action potential. A slow recovery of the spike duration was observed, although an age-dependent reduction in duration was also observed in neurones in the contralateral nucleus. 5. Two K+ conductances, the Ca2+-dependent and the A type, decrease 1 month after axotomy and follow a similar time course of recovery to that of the reduction in action potential duration and amplitude. 6. The firing pattern of axotomized neurones undergoes profound alteration, manifested as an increase in firing duration as a response to a rectangular current pulse. Examination of these alterations reveals that the reduction in both K+ conductances is responsible for the observed changes. 7. The results are discussed within the framework of the degenerative response known to take place in the nucleus following axotomy. We hypothesize that the observed phenomena reflect an increase in intracellular Ca2+ concentration which, in turn, inactivates the Ca2+ and K+ conductances. Furthermore this rise in intracellular Ca2+ may eventually be responsible for cell death.

Action Potentials↗
PubMed1987

Time course and distribution of motoneuronal loss in the dorsal motor vagal nucleus of guinea pig after cervical vagotomy.

Cells in the dorsal motor vagal nucleus (DMVN) of the adult guinea pig were counted at different times after unilateral cervical section of the vagus nerve. The counts were made from serial 30 microns coronal sections throughout the DMVN in normal and operated animals. There are three types of cells in the DMVN of guinea pig: medium-sized motoneurons that are retrogradely filled by HRP from the site of the vagotomy, small neurons, and glial cells. An interesting observation was a change in distribution of cells in the DMVN with age in unoperated guinea pigs. Following vagotomy degeneration was seen only in the motoneurons. Disappearance of motoneurons was slow and only 27% were present after 1 year. During that time the decrease in the total number of motoneurons was exponential with a time constant of 8.6 months, but degeneration in different parts of the nucleus was not uniform. Thirty-four percent of motoneurons in the caudal area of DMVN disappeared in the first month after vagotomy, while the rostral area was almost unchanged. The rostral area, however, showed rapid degeneration between 3 and 6 months after vagotomy. The central part of the nucleus degenerated at a constant rate between those of the rostral and caudal regions. At the end of 1 year, cell loss in all parts of the nucleus was approximately equal. Surviving motoneurons showed morphological changes: rounding of the soma, continuous reduction of the cell volume, and shrinkage of the nucleus. Occasional abnormal forms showing vacuolization or invaginated nuclei were seen. Calculations show that the process of degeneration lasts 25 days on the average. The marked degeneration found in dorsal vagal motoneurons, in contrast to recovery from axotomy in somatic motoneurons, is similar to that found in intrinsic neurons of the central nervous system. The slow and continuous time course of disappearance of motoneurons after vagotomy, however, is exceptional. It is reasonable to postulate that the increased vulnerability of these motoneurons may be sufficient to result in degeneration in response to what are normally nonpathological metabolic demands.

Animals↗
PubMed1987

Mapping calcium transients in the dendrites of Purkinje cells from the guinea-pig cerebellum in vitro.

1. A 10 X 10 photodiode array was used to detect stimulation-dependent absorbance changes simultaneously from many positions in the dendrite field of guinea-pig Purkinje cells which had been injected with the calcium indicator Arsenazo III in thin cerebellar slices. Signals from each element of the array were matched to positions on the cells by mapping them onto fluorescence photographs of Lucifer Yellow which had been co-injected into the cells with the Arsenazo III. 2. In response to intrasomatic stimulation the rising phase of the absorbance signals corresponded in time with the calcium spikes recorded with an intracellular electrode. There was no increase in absorbance during bursts of fast sodium spikes. Absorbance signals persisted after the sodium spikes were blocked by tetrodotoxin (TTX). In addition, the signals were largest at 660 nm and small signals of opposite polarity were found at 540 nm. These results indicate that the absorbance signals came from calcium entry into the cell resulting from the turning on of voltage-dependent calcium conductances. 3. In these experiments signals were usually seen all over the dendritic field and were weak or totally absent over the soma. In some cases signals were seen over a more restricted area. With a spatial resolution of 25 microns we were not able to see any evidence for highly localized sites of calcium entry. 4. Sometimes the rising phase of the calcium signals was separated by almost 13 ms in different parts of the dendritic field, too long to be explained by active propagation delay. This suggests that calcium spikes causing these signals can be evoked separately in different regions of the Purkinje cell dendritic field by long-lasting potentials which may reach local threshold at different times. 5. Calcium signals resulting from slow plateau after-potentials and the calcium spikes produced by them were also detected in all locations in the dendritic field. The relative distribution of amplitudes from these plateau signals was different from the distribution of evoked signals during current injection. 6. Climbing fibre synaptic activation produced calcium signals which were distributed over the dendritic arborization, but larger at the main dendritic tree where most of the synaptic contacts are located. 7. Calcium signals were also detected from the dendrites of other neurone types in the in vitro slice preparation. Thus, it is likely that these kind of measurements can be used to analyse the electroresponsiveness of many kinds of neurones in the mammalian brain.

Action Potentials↗
PubMed1986

Sodium-dependent regenerative responses in dendrites of axotomized motoneurons in the cat.

Ten days after extradural axotomy, partial spikes are found in greater than 20% of cat L7 motoneurons, while 15-21 days after axotomy the incidence increases to 60%. These responses are produced in excitable (hot) spots in the dendrites by synaptic excitation. Intracellular injection of QX-314, a lidocaine derivative and effective blocker of Na+ channels from within neurons, results in elimination of partial spikes before blocking somadendritic spikes. The action of QX-314 does not depend on changes in passive membrane properties or on changes in synaptic properties. Injections of Cs+ and Cl- ions rule out any major role for calcium, potassium, or chloride currents in the production of partial spikes. The partial spikes represent an unusual Na+-dependent dendritic phenomenon induced by axotomy when carried out relatively near the soma. It is reasonable to postulate that the partial spikes result from a higher concentration of Na+ channels in the dendrites. This may be the consequence of a high rate of production of Na+-channel proteins that are intended for the cut end of the axon; alternatively, they may result from the reflection from the cut end of such proteins produced at either a normal or an increased rate. These aberrant channels are inserted into somatic and dendritic membranes in higher concentrations than normal and, as well as producing local dendritic regions of low safety factor responsible for the partial spikes, also produce somadendritic spikes of unusually fast rise time and lower than usual threshold, which are relatively resistant to QX-314.

Action Potentials↗
PubMed1985

Intracellular injection of acetylcholine blocks various potassium conductances in vagal motoneurons.

Injection of acetylcholine into cholinergic neurons of the dorsal motor nucleus of the vagus induced membrane depolarization, an increase in input resistance, a decrease of early and late afterhyperpolarizations and a prolongation of the action potential. These effects were reversible and within 10-20 min almost complete recovery was always observed. Externally applied acetylcholine, even with doses as high as 15 mM, was not effective. Acetylcholine appeared to block voltage- and Ca2+-dependent K+ conductances. This block was manifested by the reduction of both the early and late afterhyperpolarizations and a decrease of the delayed rectification. The reversal potential for the conductance decrease was 15-30 mV negative to the resting potential. As a result of this blockade an increased Ca2+ current ensues, which is responsible for most of the prolongation of the action potential. The same responses were obtained after the injection of carbamylcholine, neostigmine and choline. However, unlike acetylcholine no sign of recovery was observed. In fact injection of neostigmine, carbamylcholine or neostigmine, together with acetylcholine, produced a delayed response which may reflect the accumulation of endogenous acetylcholine.

Acetylcholine↗
PubMed1983

Depolarizing action of GABA (gamma-aminobutyric acid) on myelinated fibers of peripheral nerves.

The inhibitory neurotransmitter GABA (gamma-aminobutyric acid) has been shown to have a depolarizing action on myelinated axons of both mammalian and amphibian peripheral nerves. In initial in vivo observations intravenous injections of GABA caused an increase in the excitability of the low-threshold, fast conducting fibers of the superficial radial and median nerves of the cat. Similar, graded, reversible effects were confirmed (using changes in the amplitude/integral of the stimulus-evoked A-fiber submaximal compound action potential to assess excitability) in in vitro studies with the isolated, desheathed frog sciatic nerve. GABA caused a mean maximal increase in half-maximal action potential of 29.8% (S.E. +/- 2.7), with an ED50 value of 0.09 mM and Hill coefficient of 0.70. This effect did not appear to desensitize, and could be reversibly antagonized by both bicuculline and picrotoxin. Comparison of agonist sensitivities showed a rank order of potency with muscimol greater than 3-aminopropanesulfonic acid greater than GABA greater than beta-guanidinopropionic acid greater than imidazole-acetic acid greater than guanidoacetic acid greater than delta-aminovaleric acid. With structure activity analysis the maximal activity was found to be related to N+-C separation near the 5 A value. Partial substitution of chloride ions in the superfusate by isethionate reversibly depressed the effect of GABA. These observations support the conclusion that extrasynaptic receptors for GABA are present on the myelinated axons of peripheral nerves.

Animals↗
PubMed1983

Mediobasal hypothalamic neurons are excited by the iontophoretic application of sodium.

In the course of studies on the responsiveness of mediobasal hypothalamic neurons to the iontophoretic application of cortisol, it was found that positive currents applied to a sodium chloride (1 M) barrel alone, but not to a choline chloride (1 M) barrel, frequently increased the firing of these neurons. Subsequently, systematic examination demonstrated that out of 102 MBH neurons 52 (51%) increased their firing by at least 30% with application of NaCl, using currents no greater than 10 nA. No such effect was obtained in response to Na application from a dilute solution (0.05 or 0.1 M). When glutamate was absent from the electrodes, the incidence of Na+ sensitivity fell to 17%, despite the routine use of backing currents to the glutamate barrel. K+ ions were more active than Na+ ions in producing excitation. When Na+ sensitivity was found, however, Na+ effects were produced by currents greater than K+ currents producing equivalent excitation. Like glutamate, K+ ions were capable of greatly enhancing responses to Na+. Comparison was made between cortisol and Na+ sensitivity in 70 MBH neurons; 28 cells responded to both, and 24 of them were inhibited by cortisol. Thus Na+ sensitivity is a frequent characteristic of MBH neurons inhibited by cortisol, and was present in 83% of cortisol-sensitive cells in this region. Iontophoresis of Na+ is commonly used as a control in pharmacological studies of the nervous system. Even more common is the case of concentrated NaCl solutions for recording. These procedures may not be as inert as previously thought, particularly in the hypothalamus.

Animals↗
PubMed1983

The effects of iontophoretic application of putative neurotransmitters on the electrical activity of rat mediobasal hypothalamic neurons in relation to their steroid sensitivity.

In order to characterize the neurons responsible for ACTH release and the suppression produced by adrenocortical steroids, we have studied the pharmacological sensitivity of neurons in the region containing the highest concentration of corticotropin releasing factor (CRF) in the brain and probably responsible for most of these activities. The effects of 8 putative neurotransmitters applied iontophoretically to more than 400 mediobasal hypothalamic (MBH) neurons in the rat were examined, and compared with cortisol sensitivity of the same neurons. Glutamate was the only agent that produced excitation exclusively, while GABA, serotonin, glycine and dopamine were inhibitory in action. Mixed excitation and inhibition were produced by histamine, acetylcholine (ACh) and norepinephrine (NE). All 8 cells excited by ACh were inhibited by cortisol and constitute only about 10% of neurons sampled; these neurons are considered to be strong candidates for CRF-releasing cells. On 7 of these neurons histamine was strongly excitatory and its action greatly outlasted the iontophoretic application. Histamine, however, excited many other neurons, including those neither excited by ACh nor steroids.

Acetylcholine↗
PubMed1981

Modification of responses to sensory and hippocampal stimuli in neurons of the rat mediobasal hypothalamus in the presence of iontophoretically applied cortisol.

The effects of iontophoretic cortisol on evoked activity was studied in rat tuberal hypothalamic units. Evoked activity from visual, auditory, sciatic nerve and hippocampal stimulation was examined in same neurons both before and during local cortisol administration. Hippocampal stimuli were studied in isolation and as conditioning stimuli for one of the sensory modalities. A constant background glutamate current was frequently used in order to shorten observation periods and to enhance inhibition of firing. Even though the predominant effect of cortisol on spontaneous firing was to reduce the firing rate, the effect of the hormone on evoked responses of all kinds was to reduce the size of the response, independent of sign, and even to reverse the nature of the response, from control facilitation to inhibition or from control inhibition to facilitation. Evoked responses in many units whose spontaneous activity was not affected by cortisol were altered by hormone. Of particular interest are a group of cells which are normally silent and respond to afferent stimulation with inhibition. The responses in these cells are reduced or changed to excitation in the inhibition. The responses in these cells are reduced or changed to excitation in the presence of cortisol and they may represent local inhibitory interneurons. The findings are related to the negative feedback of adrenocorticosteroids on CRF and ACTH release.

Animals↗
PubMed1981

Internal cesium ions block various K conductances in spinal motoneurons.

Conventional intracellular recording with low resistance electrodes was used to examine the effects of iontophoretic injections of Cs+ ions (30-200 nA for 30-500 s) into spinal motoneurons of cats anesthetized with pentobarbital and paralyzed with gallamine. The most striking effects of internal Cs+ were a great prolongation of the falling phase of action potentials, a large reduction in the amplitude of their afterhyperpolarizations, and a considerable increase in the size of delayed depolarizations. A reduction of resting membrane conductance (up to half of control values) and a small increase in membrane potential usually were evident. Although the rate of rise and amplitude of spikes sometimes were increased, the above effects on membrane properties usually were accompanied by block of antidromic invasion or synaptic spike generation, and inactivation of directly evoked spikes. Recovery of spike genesis was very rapid but the prolongation of spikes and other effects of Cs+ lasted 4-35 min, depending on the amount of Cs+ application. Larger injections of Cs+ resulted in greater depolarizations of up to 13 mV. It is concluded that internal Cs+ ions block voltage-dependent K+ conductance of spike repolarization, the Ca2+-activated K+ conductance responsible for the afterhyperpolarization, and some of the K+ conductance responsible for the resting potential. It is suggested that the enhanced delayed depolarization may result from a Cs+-blockade of an early outward K+ current which would unmask an inward current of Ca2+ ions.

Action Potentials↗
PubMed1980

Post-synaptic conductance increase associated with presynaptic inhibition in cat lumbar motoneurones.

1. Motoneurones were examined in which low-intensity p.b.s.t conditioning volleys caused a 5% or greater decrease of gastrocnemius monosynaptic e.p.s.p.s without evidence of long-lasting i.p.s.p.s on superimposed single sweeps. 2. Short constant current pulses were injected into these cells and in twenty-two of twenty-three cases the voltage decay was faster when preceded by the same p.b.s.t. conditioning stimuli which caused a decrease in the Ia e.p.s.p. 3. Comparing these decays to short pulse decays generated in a simple analogue neurone model suggested that after conditioning stimuli a tonic conductance increase had occurred which was located electrotonically remote from the soma in some cases or more diffusely in other cases. 4. Long-lasting i.p.s.p.s were brought out by averaging the baseline following conditioning stimuli in ten of fifteen cases, also suggesting a post-synaptic conductance increase. 5. Averaging the voltage response to long saturating constant current pulses showed a decreased motoneurone input resistance in three of eight cases. 6. The semilogarithmic decay of four of eleven conditioned e.p.s.p.s was more rapid than controls. 7. Although short pulse voltage decay analysis revealed consistent evidence for increased post-synaptic conductance following conditioning stimuli, it was not possible to decide if the location and extent of this conductance increase were sufficient to rule out presynaptic inhibition.

Animals↗
PubMed1980

The reversal potential as a diagnostic tool in transmitter identification.

The three-ion test still appears to be the best electrophysiological tool available to define the molecular mechanisms involved in activation of a given ionophore. New insights into synaptic mechanisms require the investigator to be alert to possible exceptional cases. In general, the treatment of the exceptional case is validly accomplished once the diagnosis is made. It is important to determine the Nernstian behavior of the reversal potential and to identify the ions that participate in the synaptic process, as well as to determine whether the conductances for these ions increase or decrease. Noise analysis, although measuring properties of the synaptic ionophore, seems the most promising physiological tool for identification of the transmitter recognition moiety of the receptor, since the open time appears to be a function of binding. It remains to develop stringent methodology for analyzing channel open time during the physiological event.

Animals↗
PubMed1979

Stoichiometry of GABA-receptor interactions: GABA modulates the glycine-receptor interaction allosterically in a vertebrate neuron.

1. Measurement of steady-state conductance changes provides a reliable method for determination of transmitter-receptor stoichiometry in general and for GABA and its receptor interactions in particular. Log-log plotting of steady-state conductance changes as a function of decreasing transmitter concentration gives the molecularity of the interaction as a limiting slope. 2. Suitable measurements of GABA action in locust muscle show a molecularity of 3 with strong positive cooperativity. One molecule of picrotoxin is sufficient to block this reaction. Kinetic studies reveal the presence of occult desensitization. Thermodynamic studies reveal strong negative heats of interaction compatible with conformational changes in a multi-subunit receptor. 3. Measurements in the lateral dendrite of the goldfish Mauthner cell reveal that glycine has a more powerful action than GABA. Both interactions, however, utilize 4 molecules of amino acid with strong positive cooperativity to activate its receptors. The receptors are apparently distinct and there appears to be a higher glycine-receptor density. 4. In addition to its action on its own receptor, GABA allosterically modulates the glycine-receptor interaction in the Mauthner cell by lowering the energy barrier for the binding of the first glycine molecule, thereby increasing the affinity of glycine for its receptor.

Allosteric Regulation↗
PubMed1979

Intracellular divalent cations and neuronal excitability.

Itracellular injections of Mg into cat spinal motoneurones have a depolarizing action, associated with a fall in input conductance, and depression of the postspike hyperpolarizing after-potential (a.h.p.) as well as its underlying conductance increase. There is also an increase in excitability, sometimes leading to outright discharge, and a change in the current-firing relation: the normal primary range is largely abolished and the firing appears to have the characteristics of the normal secondary range. Intracellular effects of Mg are thus mainly opposite to those of Ca, possibly owing to competition at sites where Ca activates K channels. Intracellular injections of Mn also tend to depress the a.h.p. but have relatively little effect on resting potential and conductance, or action potentials. Co also depresses the a.h.p. but has a more pronounced depolarizing action, and produces particularly strong depression of action potentials. By contrast intracellular Sr tends to raise the membrane conductance and has a mild hyperpolarizing effect. During the injection of Sr, a.h.p's are depressed but this is followed by a rebound of increased a.h.p. amplitude and conductance. Unlike the other divalent cations tested, Sr strongly depressed excitatory postsynaptic potentials. In most respects Sr appears to behave like Ca.

Action Potentials↗