Biomedical subjects

E F Barrett

Publications and source records attributed to E F Barrett.

26 records · Page 2Linked to original sources

PubMed1980

Voltage-sensitive outward currents in cat motoneurones.

1. The soma membrane of cat motoneurones was voltage-clamped in vivo using intracellular current and voltage electrodes whose tips were separated by at least 5 micrometer. 2. Depolarization activates two separate, non-interacting K conductance systems whose rates of activation and decay differ by a factor of about 10. These conductances have a similar reversal potential, in the range of -6 to -21 mV (these and all subsequent voltages are expressed relative to the resting potential). Both conductances show linear 'instantaneous' current-voltage relationships. The steady-state magnitudes of both conductances increase with increasing depolarization. Neither conductance inactivates substantially during prolonged depolarizations. 3. The faster K conductance is similar to that described for squid axons and frog node. Activation begins at about +30 mV and is more than 90% complete within 5 msec of a depolarizing voltage step to +50 mV. Activation kinetics appear to be nonlinear. This fast K conductance contributes to the fast falling phase of the action potential. Following repolarization, this conductance decays with a time constant of 2-4 msec. 4. The slower K conductance activates during depolarizations of 10 mV or greater. The activation and decay of this conductance can be described by first-order exponential functions with time constants ranging from 20 to 50 msec. The slow K conductance underlies the prolonged hyperpolarization that follows motoneurone action potentials. Evidence from other studies suggests that this slow K conductance is regulated by intracellular Ca ions. 5. In addition to the two K conductance systems activated by depolarization, motoneurones exhibit another distinct conductance system that is activated by hyperpolarization. This third system has a reversal potential near the resting potential. Activation of this conductance during a hyperpolarizing voltage step can be fitted by a single exponential function with a time constant of 50-60 msec over the range -20 to -50 mV. This hyperpolarization-activated conductance accounts for some aspects of the anomalous rectification reported in cat motoneurones. 6. When the clamp circuit was turned off and the motoneurones were stimulated to discharge repetitively by depolarizing current steps, the apparent soma threshold voltage increased as the applied current (and discharge frequency) increased. 7. The basic features of the motoneurone action potential were reconstructed by simulations based on voltage clamp measurements of the voltage dependent conductance systems and previous measurements of passive membrane properties. These simulations assumed that the kinetics of the fast Na and K conductance systems in motoneurones can be described by equations of the same form as the Hodgkin-Huxley equations. These action potential reconstructions indicated that a major portion of the delayed depolarization following the action potential is attributable to capacitative currents from the dendrites...

Action Potentials↗

PubMed1980

Calcium dependence of evoked transmitter release at very low quantal contents at the frog neuromuscular junction.

1. The relationship between the rate of evoked transmitter release and the extra-cellular concentration of Ca ions, [Ca2+]o, was studied at surface neuromuscular junctions of the frog cutaneous pectoris muscle. The average quantal content of the end-plate potential was reduced to low levels by reducing [Ca2+]o and adding 2 mM-Mn2+, 45 MM-Co2+ or 10 mM-Mg2+. 2. When the motor nerve was stimulated at a low frequency (0.5--2 Hz) in 2 mM-Mn2+ or 4 mM-Co2+, the average quantal content of evoked release was proportional to the fourth power of [Ca2+]o down to the lowest measurable quantal contents, around 2--4 quanta per 1000 stimuli. Combined with previous studies, this result indicates that evoked transmitter release has a steep, nonlinear dependence on [Ca2+]0 over our orders of magnitude of evoked release. 3. Calculations predict that if evoked and spontaneous release have the same fourth power dependence on intracellular [Ca2+], then the curve relating evoked release and [Ca2+]o should become much less steep as the evoked release rate approaches the spontaneous release rate. Our observation that the relationship between evoked release and [Ca2+]o remains fourth power down to very low release rates suggests that most spontaneous quantal release does not have the same dependence on intracellular [Ca2+], or does not use the same intracellular Ca2 pool, as evoked release. 4. In 2--10 mM-Mg2+, the lowest average quantal contents were markedly higher than the fourth power prediction. This discrepancy may occur either because Mg2+ somehow elevates intracellular [Ca2+], or because Mg2+ is itself a weak activator of transmitter release. 5. Even at very low rates of evoked release, increasing the stimulus frequency to 5--50 Hz caused a progressive increase in both evoked release and the rate of 'background' quantal release during the interstimulus interval. The frequency-dependent enhancement of both evoked and background release was more pronounced in solutions containing 10 mM-Mg2+ than in solutions containing 2 mM-Mn2+.

Animals↗

PubMed1978

Excitation-contraction coupling in skeletal muscle: blockade by high extracellular concentrations of calcium buffers.

High concentrations (80 to 90 millimolar) of the calcium buffers EGTA and citrate (less than 10(-7) molar free calcium ion) reversibly block excitation-contraction coupling in intact frog skeletal fibers, but do not block caffeine-induced contractures. Solutions containing the same free calcium concentration but lower concentrations of calcium buffer (1 millimolar) do not block excitation-contraction coupling. These results suggest that excitation-contraction coupling requires the presence of calcium in a "protected" extracellular compartment, probably the transverse tubular network, and that calcium is actively transported into this compartment from the muscle cell cytoplasm.

Action Potentials↗

PubMed1978

Temperature-sensitive aspects of evoked and spontaneous transmitter release at the frog neuromuscular junction.

1. The temperature dependence of presynaptic processes involved in neuromuscular transmission was studied by rapidly increasing the temperature of cooled frog neuromuscular junctions by 4--10 degrees C using pulses from a neodymium laser. The temperature elevation was complete within 0.5 msec, and decayed back to control levels with a time constant of about 7--8 sec. 2. Temperature jumps completed before nerve stimulation increased the quantal content and decreased the latency of the end-plate potential (e.p.p.). The Q10 for e.p.p. quantal content in low [Ca2+] Ringer averaged about 3.9 over the range 1--18 degrees C. 3. Temperature jumps occurring during the synaptic delay (the interval between the presynaptic action potential and the onset of the e.p.p.) also increased the quantal content and decreased the latency of the e.p.p. These effects diminished as the onset of the temperature jump was moved closer to the expected onset of the e.p.p. Temperature jumps applied after the onset of the e.p.p. immediately accelerated the time course of the e.p.p. but did not significantly alter quantal content. These results demonstrate that the magnitude and timing of evoked release are influenced by temperature-sensitive processes that operate both during and shortly after the presynaptic nerve action potential, but are largely complete before the onset of release. 4. Temperature jumps were applied at various times during the interval between two nerve stimuli. The amplitude of the second e.p.p. decreased as the temperature jump was moved earlier in the interstimulus interval, suggesting that the rise in temperature following the first nerve stimulus accelerates the decay of facilitation. When the temperature jump was moved from 10 msec after to 10 msec before the onset of the first e.p.p., the amplitude of the second e.p.p. either decreased or showed no change. The fact that the second e.p.p. did not increase suggests that the temperature-sensitive processes that increase the quantal content of the conditioning e.p.p. do not greatly increase the facilitation following that e.p.p. 5. Temperature jumps immediately accelerated the time course of spontaneous miniature end-plate potentials (m.e.p.p.s) and increased their frequency. Experiments using slow temperature changes revealed that the Q10 for m.e.p.p. frequency in normal Ringer is about 10 over the range 10--20 degrees C. M.e.p.p. frequency was much less sensitive to temperature changes below about 10 degrees C. When the nerve terminal was depolarized by 20 mM-K+ in the presence of Ca2+, the Q10 for the rate of spontaneous release over the range 10--20 degrees C decreased to about 4, similar to the Q10 for e.p.p. quantal content. In the absence of extracellular Ca2+ the Q10 for m.e.p.p. frequency in 20 mM-K+ remained near 10. 6. The marked difference in Q10S for spontaneous transmitter release under different experimental conditions suggests that not all transmitter release uses identical mechanisms...

Action Potentials↗

PubMed1976

Separation of two voltage-sensitive potassium currents, and demonstration of a tetrodotoxin-resistant calcium current in frog motoneurones.

1. Depolarization-induced voltage and conductance changes were studied in frog montoneurones in isolated, perfused spinal cord slices. Two types of afterhyperpolarization are observed following action potentials in normal Ringer, a fast afterhyperpolarization lastin 5-10 msec and a slow afterhyperpolarization lasting 60-200 msec. Both afterhyperpolarizations are mediated by an increased K+ conductance. 2. The slow afterhyperpolarization and conductance increase underlying it are selectively and reversibly inhibited by perfusion with solutions containing low [Ca2+] (less than or equal to 0-2 nM) or the Ca2+ antagonists Mn2+ (1mM) or Co2+ (5 mM), and are enhanced by perfusion with high [Ca2+]. 3. Addition of 2-5 mM tetraethylammonium ion (TEA+) to the perfusing solution prolongs the falling phase of the action potential and abolishes the fast afterhyperpolarization, but does not inhibit the slow afterhyperolarization. 4. When the voltage-dependent Na+ current is blocked by perfusion with TTX (10-5 M), intracellularly applied depolarizing current steps evoke fast and slow hyperpolarizations with kinetics and pharmacological sensitivities similar to those of the fast and slow afterhyperpolarizations, respectively. The fast hyperpolarization is maximally activated by brief, intense depolarizations, the slow hyperpolarization by prolonged, less intense depolarizations. 5. These pharmacological and kinetic data demonstrate that in frog motoneurones the repolarization-fast afterhyperpolarization sequence and the slow afterhyperpolarization are produced by different K+ conductance systems. The fast K+ conductance activates rapidly on depolarization, decays rapidly on repolarization, and is TEA+ sensitive, while the slow K+ conducatance activates and decays more slowly and is Ca2+-dependent. 6. Motoneurones perfused with TEA+ and TEA often show a slow, regenerative depolarizing response to applied depolarizing currents. These regenerative depolarizations are probably produced by an influx of Ca2+, because they persist in isotonic CaCl2 and are blocked by Mn2+ or low [Ca2+]. The Ca2+-dependence of the slow afterhyperpolarization and the increase in slow afterhyperpolarization magnitude observed following the slow Ca2+ potentials suggest that a depolarization-evoked Ca2+ influx activates the K+ conductance underlying the slow afterhyperpolarization. 7. Motoneurones in which the slow Ca2+ and K+ conductance systems have been enhanced by high [Ca2+] or blocked by Mn2+ show altered discharge patterns in response to intracellularly applied depolarizing current steps. Perfusion with twice normal [Ca2+] (4 mM) causes montoneurones to discharge more slowly at all current intensities, and reduces the slope of the 'steady-state' frequency-current relationship. Mn2+-perfused motoneurones exhibit fairly normal high-frequency discharge at the onset of the current step, but unlike normal motoneurones, do not discharge at frequencies below 60/sec...

Action Potentials↗

PubMed1974

A note on the interaction of spontaneous and evoked release at the frog neuromuscular junction.

1. The interaction between spontaneous miniature end-plate potentials and evoked end-plate potentials was investigated at the frog neuromuscular junction using focal extracellular recording techniques.2. End-plate potentials evoked immediately after a spontaneous miniature potential were facilitated by up to 20%. The percentage facilitation was negatively correlated with the average quantal content of the end-plate potential.

Animals↗

PubMed1972

Quantal independence and uniformity of presynaptic release kinetics at the frog neuromuscular junction.

1. Amplitude and latency fluctuations of the end-plate potential at the frog neuromuscular junction were studied simultaneously at low temperatures, using intracellular or focal extracellular recording techniques and average quantal contents between 0.5 and 3.2. At the release rates studied, the evoked release of one quantum has in most cases no significant effect on the probability of subsequent quantal release to the same stimulus, confirming the mutual independence of quantal releases in this preparation.3. An equation derived from Poisson's law was applied to a histogram of the latencies of the first quantum released on each of a series of trials, to predict the average quantal content of end-plate responses originating at various times after nerve stimulation. The shape of the predicted time distribution of quantal contents usually agreed closely with that of the experimentally observed time distribution of end-plate response amplitudes. This agreement demonstrates that both the amplitude and the latency fluctuations of the end-plate response result from one presynaptic stochastic process that is uniform in magnitude and time course after each stimulus.4. Analysis of extracellular records from synaptic regions with a history of extensive activity often suggested the existence of depressive interaction among quantal releases, perhaps caused by depletion of the supply of releasable quanta.

Animals↗

PubMed1972

The kinetics of transmitter release at the frog neuromuscular junction.

1. Fluctuations in the latency of focally recorded end-plate currents were analysed to determine the time course of the probabilistic presynaptic process underlying quantal release evoked after single nerve stimuli at the frog neuromuscular junction.2. The early falling phase of the presynaptic probability function can be fitted by a single exponential over two orders of magnitude of quantal release rate. The time constant of the early falling phase is about 0.5 msec at 11 degrees C, and increases with decreasing temperature with a Q(10) of at least 4 over the range 1-12 degrees C.3. After this early exponential fall, quantal release probability returns to control levels with a much slower time course.4. Conditioning nerve stimuli increase the magnitude and slightly prolong the early time course of release evoked by a test stimulus. When facilitation is calculated for matched time intervals following the conditioning and testing stimuli, it is found that the magnitude of the small, late residual tail of release is facilitated by a greater percentage than the magnitude of larger, early portions of release.5. These results are discussed in terms of the hypothesis (Katz & Miledi, 1968) that evoked release and facilitation are mediated by a common presynaptic factor which activates release in a non-linear manner.

Adaptation, Physiological↗