Electrogenesis of muscular fibers in the rat masseter muscle proper

It was established by the method of intracellular lead of membrane rest potentials (MRP) and action potentials (AP) that fibers with high MRP and little overshot are mainly located in superficial layers of the rat masseter muscle proper whereas those with low MRP and high overshot principally in deep layers. Excitability of the cytoplasmic membrane of muscular fibers of both types proved to be related in electrical properties (critical level of depolarization, current threshold). It is suggested that the rat masseter muscle contains a great number of rapid phasic fibers in superficial layers and slow ones in deep layers.     

The action of ultrasound on the contraction strength and action potential of the papillary muscle of the rat heart

In experiments on isolated rat papillary muscles the effects of therapeutic doses of ultrasound (US) (intensity, less than 2 W/cm2) with frequency of 0.88 MHz on contraction force and action potential (AP) were studied. 12 muscles (from 14) responded to 3-min exposition of the US with a rise both in contraction force and in resting tension. Sensitivity to US and a value of inotropic effect changed significantly between the preparation, and the threshold intensities of US varied from 0.3 to 2 W/cm2. In 3 experiments the inotropic effect of US was more than 100%, but in others it was about 50%. Two preparations were not sensitive to the US. The positive inotropic effect of US was accompanied by membrane depolarization (up to 20 mV) and by prolongation of AP duration measured at 10% of its amplitude (APD10). The correlation between the increase in contraction force and APD10 was demonstrated. Some preparations responded to US with high depolarization (up to 50 mV) and were inexcitable. The US induced an increase in temperature less than 1 degree C, therefore all the effects of US could not be explained as a result of temperature rise.     

The effects of the pathogenic influence of botulin toxin on different types of spinal motor neurons]

The motor neuron cells of the lumbosacral region were investigated in the spinal cord of cat with the local botulin paralysis. Development of this paralysis was followed by reduction of the membrane potential, of the amplitude of antidromic AP, mono- and polysynaptic EPSP, a fall of the input resistance and by an increase in the level of the critical depolarization of the somatic membrane of the phasic motor neurons of the damaged segments in the spinal cord. Excitation of the tonic motor neurons was not greatly altered in the dynamics of local botulism.     

Effect of a new beta-blocker (CM 6805) on the action potential of guinea pig atrial fiber

CM6804 effect has been studied about some parameters of the intracellular action potential (AP) of guinea pig autorythmic auricle. Auricle was preserved alive under Tyrode oxygenated solution at 37 degrees C. AP is measured by a fluctuating intracellular glass microelectrode. At a concentration of 5 . 10(-5) M, CM doesn't alter the resting membrane potential, it causes a small overshoot reduction, it decreases the maximum depolarization rate and the heart rate, it increases the action potential duration. Overshoot and maximum depolarization rate decrease prove that CM modifies the membrane permeability probably by a diminution of the sodium rapid inward current. CM action is similar to others aryl-oxy-propyl propanolamine like propranolol.     

A mathematical model of a myelinated nerve fiber for analysis of impulse conduction mechanisms during the relative refractory period

It was shown by means of a mathematical model of a myelinated nerve fiber (Frankenhaeuser — Huxley) that an increase in threshold and decrease in the amplitude of the action potential (AP) during the relative refractory period are due mainly to sodium inactivation. The contribution of increased potassium permeability to these changes is small, for the chief component of the outgoing ionic current in the node of Ranvier is not the potassium current, but the leak current. Given the ratio between these currents the increase in threshold and graduation of the action potential in the node membrane are less marked than in the membrane of the squid giant axon. At the beginning of the relative refractory period the AP evoked by strong stimulation is conducted only to the next node. Later in the refractory period impulses are conducted incrementally, and the threshold for the spreading impulse is higher than the threshold for spike excitation in the stimulated node. Delay in impulse conduction between refractory nodes leads to the formation of a retrograde depolarization wave. The reasons for differences in the mechanisms of impulse conduction along unmyelinated and myelinated refractory fibers are discussed.Vishnevskii Institute of Surgery, Academy of Medical Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 4, No. 2, pp. 201–207, March–April, 1972.     

ATP induces guinea pig gallbladder smooth muscle excitability via the P2Y4 receptor and COX-1 activity

The purpose of this study was to elucidate the mechanisms by which ATP increases guinea pig gallbladder smooth muscle (GBSM) excitability. We evaluated changes in membrane potential and action potential (AP) frequency in GBSM by use of intracellular recording. Application of ATP (100 microM) caused membrane depolarization and a significant increase in AP frequency that were not sensitive to block by tetrodotoxin (0.5 microM). The nonselective P2 antagonist, suramin (100 microM), blocked the excitatory response, resulting in decreased AP frequency in the presence of ATP. The excitatory response to ATP was not altered by pyridoxal-phosphate-6-azophenyl-2,4-disulfonic acid (30 microM), a nonselective P2X antagonist. UTP also caused membrane depolarization and increased AP frequency, with a similar dose-response relationship as ATP. RT-PCR demonstrated that the P2Y(4), but not P2Y(2), receptor subtype is expressed in guinea pig gallbladder muscularis. ATP induced excitation was blocked by indomethacin (10 microM) and the cyclooxygenase (COX)-1 inhibitor SC-560 (300 nM), but not the COX-2 inhibitor nimesulide (500 nM). These data suggest that ATP stimulates P2Y(4) receptors within the gallbladder muscularis and, in turn, stimulate prostanoid production via COX-1 leading to increased excitability of GBSM.     

Effects of Serotonin on Primary Afferents in the Frog Spinal Cord

We studied, on isolated preparations of the frog spinal cord, the effects of serotonin in different concentrations on the amplitude-temporal parameters of action potentials (AP) in primary afferent fibers, on the potentials reflecting depolarization of primary afferents (DPA), and on the properties of the membrane of these fibers. It was demonstrated that in a part of the dorsal root afferent fibers serotonin caused a drop in the AP amplitude (by 15-20%) and an increase in the AP duration (by 8-13%). Serotonin also significantly (by 70-90%) decreased the amplitude of DPA induced by stimulation of a neighboring dorsal root and noticeably reduced the input membrane resistance of afferent fibers. Serotonin-induced modulation of the AP parameters in the afferents and suppression of DPA under the influence of this amine are postulated as possible factors involved in the central control of afferentation.     

Mechanoelectrical feedback in the healthy heart and in the heart with pathologies

The article discusses the issues of possible connection between mechanical phenomena in myocardium and the electrical processes. Not only cardiomyocytes, but also cardiac fibroplastic are considered as substratum for the mechanisms of mechano-electrical feedbacks. Cardiomyocytes and fibroplastic of healthy animals demonstrate the mechano-electrical feedbacks, which essentially mean that stretching of the cardiac tissue within the physiological limits to 2 mN changes the electrophysiological cell processes. Close to 90% repolarization potential of cardiomyocytes action the mechano-induced depolarization develops; over the background of depolarization, when it reaches the threshold values, extra potentials of action are generated. In fibroplastic, membrane mechano-induced hyperpolarization develops; as result of cellular interaction it may develop hyperpolarization of pacemaking cells of the right auricle and slow the cardiac rythm down. In case of a pathology, for instance, infarct of the left heart ventricle modification of electric cell activity was detected at quite low values of tissue stretching up to 0.2. mN. Mechano-induced depolarization of cardiomyocytes of animals affected by infarct develops at 50% level of repolarization phase of action potential, or at 90% of repolarization phase. In the former case development of mechano-induced depolarization coincides with the period of absolute cell refractering. Extra action potential develops immediately after the refractering phase when the mechano-induced depolarization shifts the membrane potential towards threshold values. In the latter case the mechano-induced depolarization transforms into extra action potential. With further stretching fibrillation develops. In fibroplastic the values of mechano-induced membrane hyperpolarization grow with greater scope of infarct damage. Magnitude of mechano-induced hyperpolarization of auricle fibroplastic taken from the animals with infarcts shows dependence on the period of remodelling if stretching is tissue is a standard parameter. With prolongation of the remodelling period the value of mechano-induced fibroplastic hyperpolarization diminishes. The problem of developing the combinations eliminating mechano-induced cardiac arrhythmia is raised.     

Vagal and sympathetic effects on the pacemaker fibers in the sinus venosus of the heart

1. Action potentials from sinus venosus and auricle fibers of spontaneously beating frog hearts have been recorded with intracellular electrodes. 2. Sinus fibers show a slow depolarization, the pacemaker potential, during diastole. The amplitude of this potential varies in different parts of the sinus. In some fibers the membrane potential falls by 11 to 15 mv. during diastole and the transition to the upstroke of the action potential is comparatively gradual. In other regions the depolarization develops more slowly and the action potential takes off more abruptly from a higher membrane potential. It is proposed that the fibers showing the largest fall in membrane potential during diastole are the pacemaker fibers of the heart, and that the rest of the preparation is excited by conduction. In auricle fibers the membrane potential is constant during diastole. 3. The maximum diastolic membrane potential and the overshoot of the action potential vary inversely with the amplitude of the pacemaker potential. The highest values were measured in auricle fibers. 4. Stimulation of vagi suppresses the pacemaker potentials. While the heart is arrested the membrane potential of the sinus fibers rises to a level above the maximum diastolic value reached in previous beats. In 28 experiments vagal stimulation increased the membrane potential from an average maximal diastolic value of 55 mv. to a "resting" level of 65.4 mv. The biggest vagal polarization was 23 mv. 5. In contrast to the sinus fibers vagal inhibition does not change the diastolic membrane potential of frog auricle fibers. 6. Vagal stimulation greatly accelerates the repolarization of the action potential and reduces its amplitude. These changes were seen both in the sinus and in auricle fibers stimulated by direct shocks during vagal arrest. 7. The conduction velocity in the sinus venosus of the tortoise is reduced by vagal stimulation. Block of conduction often occurs. 8. In the frog sinus venosus sympathetic stimulation increases the rate of rise of the pacemaker potential, accelerating the beat. The threshold remains unchanged. The rate of rise of the upstroke and the amplitude of the overshoot are increased. 9. The analogies between the vagal inhibition of the heart and the nervous inhibition of other preparations are discussed.     

Microelectrophysiological analysis of the fiber components of the rat digastric muscle

Membrane potential at rest (MP), action potential (AP), critical level of depolarization (CLD) and latent period (LP) of different muscle fibers were studied in two bellies of digastric muscle. Even and chaotic distribution of different muscle fibers was observed in the anterior and posterior belly, respectively. It is believed that electrophysiological data correspond to the results of histological analysis of muscle fibers in digastric muscle.     

The action of the neurotoxins 5,6-dihydroxytryptamine and p-chlorophenylalanine on the electrical activity parameters of the command neurons during long-term sensitization and learning in the snail]

The influence of 5,6-dihydroxytryptamine (5,6-DHT), which selectively destroyed serotonin terminals, and p-chlorphenylalanine, which inhibited serotonin synthesis, was studied on the long-term sensitization (LTS) in a snail. The membrane mechanisms were analyzed by measuring electrical characteristics of command neurons of defensive behavior LPa3, RPa3, LPa2, and RPa2. Snails injected with saline served as an active control. It was shown that the injected drugs inhibited the LTS in certain concentrations. A significant increase was observed in the membrane potential and the threshold of the action potential generation in the command neurons after 5,6-DHT injection in the doses of 20 and 30 mg/kg (in comparison with the active control). Sensitization of snails injected with saline solution led to the LTS and decrease in the membrane and threshold potentials of the command neurons. After the LTS, changes in membrane and threshold potentials in snails injected with 5,6-DHT were negligible in comparison with those injected with 5,6-DHT but without the LTS. Neither the LTS nor subsequent learning resulted in a further decrease in membrane and threshold potentials. Thus, the neurotoxin injection led to an increase in excitability of command neurons and their depolarization, and the LDS did not elicit further excitability increase. Since the shifts of the threshold and membrane potentials were the same, it was concluded that the increase in membrane excitability was induced by the depolarizing shift of the membrane potential.     

Ion mechanisms of the mechanoelectrical feedback in myocardial cells

This article is dedicated to the mechanism of mechano-electric feedback in heart. The evidence is briefly discussed on organ, tissue, cell and in details on cell membrane levels in case of application of one of applied mechanical stimulus to cardiomyocytes. Stretch of the hole heart or its tissue fragment causes quick starting repolarization of action potentials (AP)/monophasic action potentials (MAP), shift of AP/MAP plato to higher negative zone, appearance of peaks of stretch-induced depolarization (SID) on final phase of AP/MAP repolarization level, which are overgrowing into extra AP/extra MAP. Mechanical events (changes in length and force of contractions) change electrical processes by means of direct influence on cell membrane via stretch activated channels (SAC). Cardiomyocytes, isolated from animal atrium and animal and human ventricular are responsible for the stretch of depolarized membrane, prolongation of AP and appearance of extra AP (extra systoles). Analysis of experiments, conducted following the patch clamp method in whole cell configuration, shows that the main cause of that mechanical events is SAC current--ISAC. During negative potential ISAC is determined by incoming into the cell sodium ions and is negative. Negative ISAC is changing final phase of AP/MAP repolarization and causes SID, which is overgrowing into extra AP (extra systoles), in case that SID exceeds threshold. Fast AP repolarization and AP plato shift into higher negative zone is related to positive ISAC (living potassium ions through SAC), activation of IK and reduction of ISAC. Activation of ISAC and arrhythmia induction require lower mechanical stimulus for hypertrophied cardiomyocytes, in comparisment to healthy ones. Hypertrophy of cardiomyocytes can lead to expression of SAC therefore increasing channel density and ISAC maximum amplitude. In this article is discussing data, acquired by means of direct measurement of conduction of single SAC on the background of mechanical stimulation of the cardiomyocytes membrane. It contains characteristics of the estimated SACs. It is shown that blocking of conduction of ions through SAC prevents mechanically induced arrhythmia in healthy and hypertrophied cardiomyocytes, which transforms the problem of mechano-electric feedback in heart from purely fundamental into clinical one.     

Postnatal development of electrophysiological properties of pacemaker cells in the rabbit

Using glass microelectrodes, the authors measured basic electrophysiological parameters of the true pacemaker cells of the rabbit in an attempt to elucidate the postnatal drop in the frequency of action potentials produced by the sinoatrial node. The average rate of slow diastolic depolarization falls markedly between birth and adulthood, while the duration of the action potentials of the pacemaker cells become prolonged. These changes explain age-determined chronotropic development: the lower rate of slow diastolic depolarization in adult animals causes later attainment of the threshold; the longer action potential also contributes to prolongation of the cycle of membrane voltage changes in the pacemaker cells. The direct role of the postnatal increase in the maximum diastolic potential value is small, owing to a concomitant increase in the threshold potential value.     

The influence of glutamic and aminoacetic acids on the excitability of the liverwort Conocephalum conicum

Intracellular microelectrode measurements revealed that a resting potential (RP), an action potential (AP) and a calcium component of AP (named voltage transient, VT) can be influenced by glutamic acid (Glu) and aminoacetic acid (glycine, Gly) in the liverwort Conocephalum conicum. In the continuous presence of 5mM Glu or 5mM Gly, the RP hyperpolarized constantly and the plants became desensitized to the excitatory amino acids (Glu or Gly). Under such circumstances, the amplitudes of APs evoked by stimuli other than Glu or Gly grew, as did their calcium components (VTs). The sudden application of 1-15 mM Glu or Gly to a thallus not yet desensitized resulted in an excitation, i.e. a single AP or AP series. Aspartate (Asp) could not substitute for Glu in any way. Simultaneous action of both amino acids acted synergically to trigger APs. The same phenomenon was observed when glycine solution was enriched with N-methyl-D-aspartic acid (NMDA). Gly-induced APs were totally hindered by 1mM D-amino-5-phosphonopentanoic acid (AP5)--an inhibitor of ionotropic glutamate receptors of the NMDA kind. Glu-induced APs could be totally suppressed by 1mM AP5 as well as by 1mM 6,7-dinitroquinoxaline-2,3-dione (DNQX)--an inhibitor of AMPA/KA receptors. DNQX also completely blocked the calcium component of Glu-evoked APs. After DNQX treatment, the only response to Glu was a membrane potential hyperpolarization (like the Glu response in a desensitized plant). It was concluded that the Glu-induced depolarization and hyperpolarization are separate phenomena. The stimulatory effects of both Glu and Gly on liverwort excitability may be the consequences of an activation of a variety of ionotropic Glu receptor subtypes.     

Neuromuscular transmission in an insect visceral muscle

The electrical properties of the muscles of locust oviduct have been examined using intracellular recordings. The muscle cells are both dye and electrically coupled. They possess a wide array of spontaneous electrical activity ranging from slow oscillations of membrane potential to action potentials. In addition to possessing spontaneous electrical activity, certain regions of the oviduct are under motor control. The amplitude of evoked excitatory junction potentials (EJPs) increased step wise revealing innervation from a maximum of three motor units. These EJPs underwent summation and facilitation, and reached a critical threshold at which point the membrane revealed an active response. Bath applied glutamate, aspartate, proctolin, and octopamine were tested for their ability to alter resting potential and EJPs. L-glutamate (1.6 X 10(-5) M and above) produced a dose-dependent depolarization of membrane potential accompanied by a reduction in amplitude of EJPs. Although L-aspartate resulted in similar effects, the concentrations required were higher than those for glutamate. Proctolin (6.3 X 10(-11) M-6.0 X 10(-9) M) resulted in a dose-dependent depolarization but had little or no effect on amplitude of EJPs. Application of D, L-octopamine (3.2 X 10(-5) M-1.7 X 10(-4) M) induced a small hyperpolarization and a reduction in amplitude of EJP. It is suggested that contractions of locust oviduct appear to be regulated by a combination of a classical neurotransmitter such as glutamate, along with the neuromodulators octopamine and proctolin.     

Altered threshold of the mitochondrial permeability transition pore in Ullrich congenital muscular dystrophy

We have studied the effects of rotenone in myoblasts from healthy donors and from patients with Ullrich congenital muscular dystrophy (UCMD), a severe muscle disease due to mutations in the genes encoding the extracellular matrix protein collagen VI. Addition of rotenone to normal myoblasts caused a very limited mitochondrial depolarization because the membrane potential was maintained by the F1FO synthase, as indicated by full depolarization following the subsequent addition of oligomycin. In UCMD myoblasts rotenone instead caused complete mitochondrial depolarization, which was followed by faster ATP depletion than in healthy myoblasts. Mitochondrial depolarization could be prevented by treatment with cyclosporin A and intracellular Ca(2+) chelators, while it was worsened by depleting Ca(2+) stores with thapsigargin. Thus, in UCMD myoblasts rotenone-induced depolarization is due to opening of the permeability transition pore rather than to inhibition of electron flux as such. These findings indicate that in UCMD myoblasts the threshold for pore opening is very close to the resting membrane potential, so that even a small depolarization causes permeability transition pore opening and precipitates ATP depletion.     

Effect of high pH on the plasma membrane potential and conductance in Elodea densa

In leaves of Elodea densa the membrane potential measured in light equals the equilibrium potential of H+ on the morphological upper plasma membrane. The apoplastic pH on the upper side of the leaf is as high as 10.5-11.0, which indicates that alkaline pH induces an increased H+ permeability of the plasmalemma. To study this hypothesis in more detail we investigated the changes in membrane potential and conductance in response to alterations in the external pH from 7 (= control) to 9 or 11 under both light and dark conditions. Departing from the control pH 7 condition, in light and in dark the application of pH 9 resulted in a depolarization of the membrane potential to the Nernst potential of H+. In the light but not in the dark, this depolarization was followed by a repolarization to about -160 mV. The change to pH 9 induced, in light as well as in dark, an increase in membrane conductance. The application of pH 11, which caused a momentary hyper- or depolarization depending on the value at the time pH 11 was applied, brought the membrane potential to around -160 mV. The membrane conductance also increased, in comparison to its value at pH 7, as a result of the application of pH 11, irrespective of the light conditions.     

Effects of the membrane potential level on serotonin-induced contraction of the pulmonary artery smooth muscle in rabbits]

The effects of changes in membrane potential level on the electrical and contractile responses induced by serotonin (10(-6) mol/l) were investigated in muscle strips from rabbit main pulmonary artery using sucrose-gap technique. In spite of the fact that serotonin-induced depolarization did not exceed the threshold level for development of contraction, it was followed by a strong tonic contraction. Nearly a half of this contraction could be relaxed by an electrotonic hyperpolarization of the membrane. A week preliminary depolarization of the muscle cells resulted in an increase while a strong depolarization--in dramatic decrease of serotonin-induced contraction. Nifedipine effectively blocked potassium-induced, but not serotonin induced contraction. We suggest that in addition to voltage-operated and receptor operated Ca channels in vascular smooth muscle cell membrane there is a separate class of nifedipine-insensitive Ca channels operated by both serotonin receptor and membrane potential.     

Effects of Polarization of the Receptor Membrane and of the First Ranvier Node in a Sense Organ

It has previously been shown that the site of production of the generator potential in Pacinian corpuscles is the receptor membrane of the non-myelinated ending, and the site of initiation of the nerve impulse, the adjacent (first) Ranvier node. Effects of membrane polarization of these sites were studied in the present work. Nerve ending and first Ranvier node were isolated by dissection, electric activity was recorded from, and polarizing currents were passed through them. All observations were done at steady levels of polarization, seconds after onset of current flow. The following results were obtained: The amount of charge transferred through the excited receptor membrane is a function of the electrical gradients across the membrane. The generator potential in response to equal mechanical stimuli increases with resting potential of the receptor membrane. The refractory state of the generator potential is not affected by polarization. The electrical threshold for impulse firing at the first Ranvier node (measured by the minimal amplitude of generator potential which elicits a nodal impulse) is nearly minimal at normal resting potential of the node. Both, hyperpolarization and depolarization lead to a rise in nodal threshold. For any level of polarization of nodal and receptor membrane, the threshold for production of impulses by adequate (mechanical) stimulation appears determined by the generator potential-stimulus strength relation and by the electrical threshold of the node.     

Effects of changes in membrane potential on the cyclosporin-induced inhibition of T-cell proliferation.

Cyclosporin A (CsA) exerts its major immunosuppressive effect by inhibition of T-lymphocyte proliferation. The precise mechanism and target of its action has not yet been completely identified. CsA is also known to induce a rapid membrane depolarization in T lymphocytes. We have tested the role of CsA-dependent depolarization in the inhibition of T-cell proliferation by the drug. In these studies, induced membrane depolarization (in the presence of gramicidin or by replacing the Na+ content of the medium with K+) or hyperpolarization (in the presence of valinomycin) had no influence on the induction of T-cell competence by phorbol dibutyrate/ionomycin or by submitogenic concentrations of PHA, a target for CsA immunosuppression. However, regardless of the state of membrane potential during the induction of T-cell competence, the inhibition by CsA was the same as seen in normally polarized cells. We conclude that the depolarization induced by CsA is not a critical element in its inhibitory effect on T-cell proliferation.