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E Gruen

Publications and source records attributed to E Gruen.

28 records · Page 2Linked to original sources

An aminopyridine-sensitive, early outward current recorded in vivo in neurons of the precruciate cortex of cats using single-electrode voltage-clamp techniques.

Studies were performed in cortical neurons to determine if voltage- and time-dependent membrane currents could be recognized and characterized in the dynamic, in vivo state. Intracellular measurements made in neurons of the precruciate cortex of awake cats with single-electrode voltage-clamp (SEVC) techniques disclosed an early outward current to depolarizing command steps in 124 of 137 cells studied. The voltage-dependent properties of the early outward current closely resembled those of A-currents studied in vitro in vertebrate and invertebrate neurons. The current was activated rapidly at onset latencies of less than two ms, fell to flat plateau levels within 60-120 ms during sustained depolarization, and was reduced or eliminated in 22 of 23 cells following intracellular administration of 3- or 4-aminopyridine. The magnitude of outward current in response to depolarizing commands was increased by preceding steady hyperpolarization and reduced by preceding steady depolarization. (The steady potentials were of 9.8 s duration and +/- 40 mV apart from the holding potentials.) Since return to the holding potentials occurred 80 ms before the onset of the command steps, the changes in membrane properties that were induced lasted beyond cessation of the steady polarizing stimuli themselves. Spiking did not prevent recognition of the early outward current as judged from its appearance before and after intracellular application of QX-314 to reduce spike activity. Apart from fast inward currents associated with spike potentials, the early outward current was the most conspicuous and characteristic membrane current noted in these recordings. An additional current component that was noted but not characterized in these studies was a slow, depolarization-induced inward current that could be reduced by intracellular injection of QX-314.

Aminopyridines↗

Acetylcholine reduces net outward currents measured in vivo with single electrode voltage clamp techniques in neurons of the motor cortex of cats.

Effects of acetylcholine (ACh) on membrane currents of cells of the motor cortex were measured directly, in vivo, in awake cats using single electrode voltage clamp (SEVC) techniques. Extracellular applications (90-95 nA) of 2 M ACh for periods of 30 s or less produced significant decreases in net outward currents elicited by depolarizing commands whereas applications of saline did not. Reductions of net outward currents were also obtained after intracellular pressure injections of cyclic guanosine monophosphate (GMP)-dependent protein kinase (cGPK) mixed with 10 microM cyclic GMP.

Acetylcholine↗

Intracellular injection of cGMP-dependent protein kinase results in increased input resistance in neurons of the mammalian motor cortex.

Purified, cyclic GMP-dependent protein kinase (cGPK) was pressure-injected into neurons of the precruciate cortex of awake cats. Input resistances increased within seconds after injection and remained elevated for 2 min or longer. The increases were larger when cGPK was injected in a mixture with 10 microM cGMP than when injected alone. Injections of heat-inactivated cGPK, with or without 10 microM cGMP, failed to produce increases in input resistance. The present results indicate that injection of activated cGPK into neurons of the mammalian motor cortex can mimic actions of extracellularly applied acetylcholine and intracellularly applied cGMP, the latter in 100-fold higher concentrations than those used here, in neurons of the same cortical areas.

Action Potentials↗

Responses of morphologically identified cortical neurons to intracellularly injected cyclic GMP.

Cyclic nucleotides are thought to act as second messengers of neurotransmission inside central neurons, and cyclic guanosine monophosphate (cGMP) has been postulated to act as a messenger for muscarinic, cholinergic transmission. Nonetheless, the action of cGMP has not yet been established in identified cortical neurons. We injected cGMP and horseradish peroxidase (HRP) intracellularly in neurons of the motor cortex of awake cats. Fifty-four percent of injected cells responded to cGMP and HRP with an increase in input resistance within 30 s after injection. None of a control group of cells injected with HRP without cGMP so responded. In cells receiving intracellular depolarizing current sufficient to produce repeated spike discharge at the time of injection, the increase in input resistance after cGMP persisted for as long as the cells could be held. There was no significant increase in firing rate after injection of cGMP. Cells responding to cGMP with an increased input resistance were identified as pyramidal cells of layer V. One inverted pyramidal cell of layer VI also showed an increase in input resistance in response to cGMP. Previous studies have suggested that muscarinic cholinergic agents produce an increased input resistance (thought to reflect a decreased potassium conductance) underlying an increased rate of discharge in neocortical neurons. Our results favor a dual action of muscarinic cholinergic transmission in mammalian cortical neurons--the increase in input resistance in layer V pyramidal cells mediated by cGMP, and the increase in rate of discharge mediated by other means.

Animals↗

Responses of morphologically identified cortical neurons to intracellularly injected cyclic AMP.

Intracellular injections of cyclic adenosine monophosphate (cAMP) and horseradish peroxidase (HRP) were made in neurons of the motor cortex of awake cats. Eighty-six percent of injected cells responded to cAMP and HRP with a rapid decrease in input resistance. The decreases in input resistance occurred immediately after injection and began to return toward baseline 2 to 3 min later. The decreases were significantly greater than the small decreases in input resistance normally seen in uninjected cells held for 2 min or more after penetration and exceeded comparably small decreases in input resistance seen after control injections of 5'-AMP plus HRP. Pyramidal cells of layer V were identified as responding to cAMP with a decreased input resistance. A spiny stellate cell of layer III and a pyramidal cell of layer VI were also identified that showed similar responses. Increased rates of discharge were also observed after penetration with electrodes containing cAMP, but significant changes in input resistance were not found in association with the increased rates of discharge. After pressure injection of cAMP, the rates of discharge decreased toward more normative values. Our findings indicate that cAMP has an effect on cortical neurons similar to that found in some types of invertebrate (molluscan) neurons and dissimilar to the effect of cyclic guanosine monophosphate.

Animals↗

Two different mechanisms control inhibition of spike discharge in neurons of cat motor cortex after stimulation of the pyramidal tract.

Intracellular recordings were made from neurons of the motor cortex of awake cats while the pyramidal tract (PT) was stimulated at the level of the facial nucleus. In some neurons IPSPs of 35-120 ms peak latency were recorded that diminished in size or reversed with hyperpolarizing current. During these IPSPs a decrease in input resistance reflective of a conductance increase was measured. More often, however, PT stimulation produced IPSPs with comparable latencies that increased in size with hyperpolarizing current. These IPSPs diminished with depolarizing current, and in some instances they appeared to reverse with strong depolarization. During these IPSPs an increased input resistance reflective of a decreased conductance was measured. The results indicate that two different mechanisms control rapid inhibition of spike discharge in neurons of the motor cortex after PT stimulation.

Animals↗

Intradendritic recordings from neurons of motor cortex of cats.

The properties of neuronal spike potentials ranging from 20 to 60 mV in size were studied in stable recordings made from the motor cortex of unanesthetized, unparalyzed cats. Histologic evidence that injection of horseradish peroxidase (HRP) was confined to single neurons and appropriately high levels of potassium measured with K+ ion-sensitive electrodes indicated that these recordings were obtained from single, intracellularly penetrated neurons. Pressure injection of small volumes of 4% HRP in 1 M KC1, intracellularly, was characteristically associated with transient increases in spike size and decreases in input resistance. Recoveries of HRP-filled dendritic processes without filling of somata were obtained from penetrations giving spikes smaller than the recorded resting potential (spike-undershoot recordings). Recordings with dendritic recoveries had higher input resistances and showed greater increases in spike size during pressure injection than did recordings with both somata and dendrites recovered. The activity in response to a weak click elicited in cells with spike potentials between 20 and 40 mV was as great or greater than that elicited in cells with larger spike potentials or in extracellularly recorded units. We conclude that many stable spike-undershoot recordings of neurons of the motor cortex of awake cats reflect penetrations of dendritic processes as opposed to injury because they show a) a normal functional response to weak auditory stimuli delivered over periods of many minutes, b) a reversible increase in spike size during pressure injection of small volumes of HRP in KC1 consistent with penetration of a cable remote from the site of spike generation, and c) the frequent recovery of HRP-marked dendrites without somata.

Animals↗

Characterization of electrophysiological properties of intracellularly recorded neurons in the neocortex of awake cats: a comparison of the response to injected current in spike overshoot and undershoot neurons.

Intracellular recordings were obtained from 212 neurons of the coronal pericruciate cortex of 7 awake, untrained cats. Glass microelectrodes, filled with K+ citrate alone or K+ citrate with either cyclic GMP or 5'-GMP were used for recording and for injecting steady depolarizing and hyperpolarizing currents intracellularly. The effects of rectangular linearly rising (ramp) current pulses were also studied. Results were compared in spike overshoot* versus undershoot recordings. Spike overshoot recordings had action potentials (APs) larger than associated baseline shifts on penetration; undershoot recordings had APs smaller than associated baseline shifts on penetration. Undershoot recordings are more commonly encountered in mammalian neocortex than are overshoot recordings. (1) Except for sizes and slopes of APs and other effects consistent with the penetration of passive dendritic cables remote from regions of active spike initiation or propagation, no differences in response to current injection or in other electrophysiological properties were found between overshoot and undershoot recordings. (2) Injection of depolarizing currents produced de-reases in the amplitudes of APs, decreased rates of rise and fall of APs and increased frequencies of AP discharge. Injection of hyperpolarizing current produced slowing or cessation of AP discharge with little or only slight increases in AP amplitude when the resting potential was greater than 47 mV. (3) An effectively linear relationship was found between changes in AP size and the magnitude of weak injection depolarizing currents. This relationship provides a basis for measuring changes in cortical neuronal input resistance by the differential spike height method. (4) Most neurons showed little or no accommodative response to the injection of linearly rising, depolarizing currents. Simple or ceiling threshold-latency curves rather than minimal gradient curves were obtained in 83% of the cells in which ramp currents were injected. (5) Modal values of resting potentials between 47 and 53 mV, without increased rates of spontaneous discharge, indicate that most cells have a critical firing threshold near that reported for somatodendritic (SD) rather than initial segment (IS) generated spikes. The evidence suggests that undershoot recordings primarily reflect penetrations of passive dendritic regions rather than functional modification of neurocellular properties as a consequence of impalement.

Animals↗

Effects of acetylcholine and cyclic GMP on input resistance of cortical neurons in awake cats.

The effects on input resistance (Rm) of extracellular iontophoresis of acetylcholine (ACh) and of intracellular iontophoresis of cyclic GMP (cGMP) were studied in neurons of the coronal-pericruciate cortex of awake cats. Control studies were also conducted including iontophoresis of saline extracellularly and 5'-GMP intracellularly. (1) Substantial increases in Rm occurred in approximately half the neurons given ACh or cGMP. (2) In the absence of associated repetitive spike discharge induced by intracellular injection of depolarizing current pulses during iontophoretic applications, the increases in Rm were transient occurring in less than 30 sec and lasting 4--5 min. (3) With associated current-induced spike discharge, the increases in Rm persisted for as long as the neurons could be held -- up to 1.5 h maximally. (4) Rm was not increased if saline was substituted for ACh, if 5'-GMP was substituted for cGMP, or if the neuron was only discharged repeatedly. (5) The magnitude and time course of both transient and persistent increases in Rm were comparable between cells given ACh or cGMP and whether action potentials and resting potentials were greater than or equal to 40 mV (average 47 mV)* or were less.

Acetylcholine↗