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The electrogenic Na+/HCO3- cotransport modulates resting membrane potential and action potential duration in cat ventricular myocytes.

Perforated whole-cell configuration of patch clamp was used to determine the contribution of the electrogenic Na+/HCO3- cotransport (NBC) on the shape of the action potential in cat ventricular myocytes. Switching from Hepes to HCO3- buffer at constant extracellular pH (pH(o)) hyperpolarized resting membrane potential (RMP) by 2.67 +/- 0.42 mV (n = 9, P < 0.05). The duration of action potential measured at 50% of repolarization time (APD50) was 35.8 +/- 6.8% shorter in the presence of HCO3- than in its absence (n = 9, P < 0.05). The anion blocker SITS prevented and reversed the HCO3- -induced hyperpolarization and shortening of APD. In addition, no HCO3- -induced hyperpolarization and APD shortening was observed in the absence of extracellular Na+. Quasi-steady-state currents were evoked by 8 s duration voltage-clamped ramps ranging from -130 to +30 mV. A novel component of SITS-sensitive current was observed in the presence of HCO3-. The HCO3- -sensitive current reversed at -87 +/- 5 mV (n = 7), a value close to the expected reversal potential of an electrogenic Na+/HCO3- cotransport with a HCO3-:Na+ stoichiometry ratio of 2: 1. The above results allow us to conclude that the cardiac electrogenic Na+/HCO3- cotransport has a relevant influence on RMP and APD of cat ventricular cells.

4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfo↗

Change in the summating potential and action potential during the fluctuation of hearing in Ménière's disease.

Summating potential (SP) and action potential (AP) were monitored by extratympanic electrocochleography (ECoG) during the fluctuation of hearing in 8 patients with Ménière's disease. The relationship of SP and AP parameters to pure-tone hearing threshold level was examined. The correlational analysis revealed the following: AP amplitude decreases with increase in the hearing threshold level at higher frequencies (2-8 kHz), whereas it alters independently of the change in the hearing threshold level at lower frequencies (0.25-1 kHz). -SP amplitude is unchanged irrespective of changes in the hearing threshold level at any frequency. The ratio of -SP to AP amplitude increases with increase in the hearing threshold level at higher frequencies (2-8 kHz), whereas it alters independently of the change in the hearing threshold level at lower frequencies (0.25-1 kHz).

Action Potentials↗

Summating potential and action potential ratio in Meniere's disease before and after treatment.

OBJECTIVE: This study aimed to compare the summating potential and action potential ratio (SP:AP) in patients with Meniere's disease before and after various surgical and medical treatments as an indication of change in endolymphatic hydrops to study the progression of Meniere's disease. STUDY DESIGN: The study design was a retrospective case review. SETTING: The study was conducted at an otology-neurotology referral center. PATIENTS: Eighty-eight ears of 84 patients with Meniere's disease received medical treatment in 18 ears and surgical treatments including endolymphatic shunt in 12 ears, streptomycin perfusion of the lateral semicircular canal in 9 ears, streptomycin perfusion of the middle ear in 33 ears, and dexamethasone perfusion of the middle ear in 16 ears. INTERVENTION: Transtympanic electrocochleography (ECoG) was performed in all patients before treatment and 1-57 months after treatment (mean, 13.5 months) with a 2-year interval in 28 patients. MAIN OUTCOME MEASURE: An enlarged SP:AP ratio (> or = 0.40) was used as the diagnostic criterion for endolymphatic hydrops. RESULTS: Overall, an enlarged SP:AP ratio was found in 75% of ears before treatment and 78% after treatment. The SP:AP ratio, when enlarged, remained enlarged in 91% of ears. A nonenlarged SP:AP ratio before treatment became enlarged after treatment in 41% of ears. In American Academy of Otolaryngology-Head and Neck Surgery stage 1 Meniere's disease (pure-tone threshold average < or = 25 dB), an enlarged SP:AP was found in 58% of ears before treatment and 79% after. Twenty-five (89%) of 28 patients followed for 2 years were free of vestibular symptoms after treatment, and in 22 patients (79%), the SP:AP remained enlarged. The distribution of an enlarged SP:AP ratio was associated with the duration of disease (chi-square = 33.5552, p < 0.01). CONCLUSIONS: The development of endolymphatic hydrops, as indicated by an enlarged SP:AP, is part of the progression of Meniere's disease. The longer the duration of the disease, the more likely the SP:AP ratio will be enlarged. These findings indicate that endolymphatic hydrops as detected by ECoG was not reversed in this study by the treatments used. Despite the absence of definitive spells of vertigo in most patients, endolymphatic hydrops as evidenced by an enlarged SP:AP ratio persisted.

Adolescent↗

Action potentials and membrane currents of isolated single smooth muscle cells of cat and rabbit colon.

Membrane potentials, action potentials and macroscopic currents in enzymatically dispersed, single smooth muscle cells of the circular layer of cat and rabbit colon were investigated. The cells did not exhibit spontaneous depolarizations and repolarizations (slow waves) or spontaneous action potentials. Single action potentials of smooth muscle cells were evoked by depolarizing current pulses of 5 ms to 3 s duration. A repetitive action potential discharge and an increase in the duration of the action potential was observed in cells during long depolarizing current pulses by superfusion with tetraethylammonium (TEA) or 4-aminopyridine (4-AP). Tetrodotoxin (TTX) did not alter the configuration of the action potential. Voltage-clamp experiments revealed two major outward macroscopic currents: a quasi-instantaneous (time-independent) and a time-dependent outward current. Both currents were identified as potassium (K) currents due to their pharmacological sensitivity to K antagonists [TEA, 4-AP and cesium (Cs)] and due to the reversal potential of outward tail currents. Barium selectively blocked the time-independent current. A time-dependent outward K current in colon cells was observed which appeared to be dependent upon entry of calcium ions (Ca2+) through voltage-dependent Ca-channels, since it was blocked by cadmium and low concentrations of nifedipine. The majority of cells did not exhibit transient outward currents. Inward currents were exposed in some of the cells when the K currents were blocked by external TEA and by replacement of K by Cs and TEA in the recording pipette. Inward currents were presumably carried by Ca2+, since they were not altered by TTX, were sensitive to external Ca concentrations and were abolished by the Ca channel antagonist, nifedipine. Carbachol augmented the amplitude of the inward Ca current.

Action Potentials↗

Summating potential and action potential gradients on and in the vicinity of the round window in guinea pig.

The purpose of this study was to determine the spatial gradients of summating potential (SP) amplitudes, action potential (AP) amplitudes, and SP/AP ratios for recording loci on the round window (RW) membrane and in its vicinity. Sixteen guinea pigs were tested by means of free-field click and tone burst (2.0 kHz) stimuli. Seven recording regions were specified: five equal-area regions on the RW membrane and two basal promontory regions. Statistically significant differences were found between the promontory regions and the RW membrane regions for the SP, AP, and SP/AP obtained with click stimuli, and for the SP and AP obtained with tone burst stimuli. The SP/AP ratio for tone burst stimuli did not differ significantly across the seven regions. The RW membrane was found to be isoelectric, but there were marked spatial gradients on the basal promontory. These results are of consequence for the interpretation of transtympanic electrocochleography recordings.

Animals↗

Long-term facilitation of excitatory synaptic transmission in single motor cortical neurones of the cat produced by repetitive pairing of synaptic potentials and action potentials following intracellular stimulation.

The effects of postsynaptic firing activity on excitatory postsynaptic potentials (EPSPs) were studied in the motor cortex of anaesthetized cats. Postsynaptic firing was induced by 1-5 nA cathodal current pulses via the recording intracellular microelectrode, while EPSPs were elicited by thalamic, callosal, pyramidal tract and somatosensory stimuli. In 102 cells, EPSP-spike stimulus pairs were applied with 0.2-1/sec frequency and 10-100 msec interstimulus intervals. In 42 neurones, reversible facilitation of paired EPSPs appeared lasting from 4 to 47 min. The synaptic facilitation in most cases was accompanied by membrane depolarization and an increase in input resistance. The effectiveness of current induced action potentials upon test EPSPs provided evidence for the postsynaptic localization of plastic changes occurring in conditioning experiments.

Action Potentials↗

Lidocaine effects on action potentials of Purkinje fibers from neonatal and adult dogs.

We studied the effects of lidocaine (L), 2 to 40 mg/l, on the cellular electrophysiologic properties of Purkinje fibers (PF) from neonatal (0-10 days) and adult dogs. Microelectrode impalements of neonatal and adult PF were selected so that there were no differences between the two groups in control maximum diastolic potential, action potential amplitude and maximum upstroke velocity of phase 0 depolarization (Vmax). However, control action potential duration was significantly shorter in neonates than in adults (P less than .05). At a superfusate [K+] = 4 mmol/1, the threshold concentration for L effects on action potential amplitude in neonates was 40 mg/1 and in adults, 20 mg/1; on Vmax in neonates threshold was 5 mg/1 and in adults was 2 mg/1; on action potential duration in neonates and adults threshold was 2 mg/1. For all variables studied the effect of L on adult PF action potentials was greater than on neonatal PF. The effects of L on action potential duration also suggest that potassium conductance may be greater in neonatal fibers. When superfusate [K+] was increased to 6 mM the effects of L on adult PF were potentiated more than on neonates. These results provide further experimental evidence of the age-related changes in sensitivity of the specialized conducting system to cardioactive drugs.

Action Potentials↗

Ciguatoxin-induced oscillations in membrane potential and action potential firing in rat parasympathetic neurons.

The actions of ciguatoxins from the Pacific (P-CTX-1) and Caribbean (C-CTX-1) regions were investigated in isolated parasympathetic neurons from rat intracardiac ganglia using patch-clamp recording techniques. Under current-clamp conditions, bath application of P-CTX-1 (1-10 nm) or C-CTX-1 (10-30 nm) caused a gradual depolarization that was accompanied by oscillation of the membrane potential leading to tonic action potential firing. Membrane potential oscillations were observed between -45 and -60 mV and had an amplitude of 10-20 mV and a mean frequency of 10 Hz. Oscillation frequency was temperature-dependent with a Q10 of 2.0. Membrane oscillations were temporarily inhibited by hyperpolarizing current pulses and potentiated by weak depolarizing current pulses. The amplitude of oscillations was reduced upon lowering the external Na+ concentration and inhibited by tetrodotoxin (TTX), tetracaine or Zn2+. Tetraethylammonium, 4-aminopyridine, Cs+, Cd2+, Ba2+, 1,4,4'-diothiocyanato-2,2'-stilbenedisulphonic acid (DIDS) and ouabain had no effect on the CTX-1-induced membrane depolarization and oscillations. Brevetoxin (PbTx-3, 100 nm), in contrast to CTX-1, caused a membrane depolarization that was not associated with oscillation of the membrane potential. Under voltage-clamp conditions, P-CTX-1 inhibited the peak amplitude of the voltage-dependent Na+ current and shifted the activation curve to more negative potentials, but membrane oscillations were not seen in this configuration. These results suggest that ciguatoxins cause oscillation of the membrane potential in mammalian autonomic neurons by modifying the activation and inactivation properties of a population of TTX-sensitive Na+ channels.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

Development of endocochlear potential and compound action potential in the rat.

The present study was designed to investigate the developmental changes of the endocochlear potential and compound action potential simultaneously from rat pups of various ages. Animals were anesthetized with ketamine/xylazine, and the endocochlear potential was measured with a glass microelectrode. At the same time, a wire electrode was placed on the round window to record the click-evoked compound action potential. The endocochlear potential was found to be very low during the first few days of postnatal life. A rapid increase in the value of the endocochlear potential was noted between eleven and thirteen days of age, and adult-like values were recorded by seventeen days of age. Compound action potential responses were recorded at thirteen days of age to high intensity clicks, followed by a progressive improvement of thresholds and reduction of latencies. The development of the endocochlear potential and compound action potential was found to be reciprocally related - as the magnitude of the endocochlear potential increased, the compound action potential threshold declined with increasing age. The development of the endocochlear potential was found to closely approximate the development of enzymatic activity of sodium, potassium-ATPase in the stria vascularis reported by Kuijpers (1974).

Action Potentials↗

The use of a microcomputer to automate measurement of action potential duration for both transmembrane and monophasic action potentials.

Measurement of action potential duration is made more valuable if it can be made simultaneously with other variables, to which it may be related. We have developed a microcomputer-based system which allows measurement of action potential duration, both for transmembrane action potentials and for monophasic action potentials. The system allows simultaneous recording and analysis of action potentials and intraventricular pressures. Both end-diastolic and maximum systolic pressures have been analysed. Action potential duration was assessed at four different levels of the repolarization curve. We have analysed the consistency of measurements made by the computer, and compared them to measurements made manually, using results from six dog experiments. For action potential duration, there was no systematic difference between the manual and the computer methods, but the computer was significantly more consistent. In the case of the pressure measurements, the two methods were approximately the same in their consistency, and again there was no systematic difference. We have demonstrated that potential errors in determination of the average diastolic potential did not significantly affect the results obtained by our method. The variances of action potential duration measurements made at different levels of repolarization were equal. We demonstrated that there was no effect of amplitude on the action potential duration of potentials recorded under steady-state conditions.

Action Potentials↗

Far-field potentials recorded from action potentials and from a tripole in a hemicylindrical volume.

There is growing evidence in support of the hypothesis that far-field potentials are recorded when action potentials encounter discontinuities in the surrounding volume. The present study found further support for this hypothesis using two methods of experimentation. The first method recorded potentials when the action potential from an isolated bullfrog sciatic nerve in a hemicylindrical volume (i) encountered a change in the shape of the surrounding volume, (ii) crossed a boundary between 2 volumes of differing resistivities, (iii) reached a bend in the nerve, or (iv) reached the functional end of the nerve. In the second method, potentials were recorded when an electrical tripole, constructed in a way to produce the electrical equivalent of an action potential, encountered the same discontinuities as well as when it was configured to simulate a curved nerve. These results are consistent with the hypothesis that dipole components of an action potential predominant in far-field recordings.

Action Potentials↗