Biomedical subjects

R W Greene

Publications and source records attributed to R W Greene.

At least 55 records · Page 3Linked to original sources

PubMed1991

Muscarinic agonists activate an inwardly rectifying potassium conductance in medial pontine reticular formation neurons of the rat in vitro.

Intracellular recordings were obtained from neurons in pontine reticular formation slices of the rat to characterize a cholinergic-gated increase in conductance. The conductance increase was associated with a hyperpolarization of the membrane potential and with an outward current under voltage-clamp conditions. Current-voltage relations and potassium substitution experiments indicated mediation by a change in permeability, primarily to potassium. This potassium conductance exhibited inward rectification at membrane potentials negative to resting potential, a novel finding for cholinergic actions in CNS neurons. Further characterization of this inwardly rectifying potassium conductance revealed marked sensitivity to low concentrations of barium. Cholinergically evoked currents were relatively unaffected by the presence of extracellular cesium. Cholinergic effects persisted in TTX. The outward currents elicited by carbachol or methacholine were blocked only by high concentrations of pirenzepine, a selective antagonist of the M1 muscarinic receptor. The interaction between these agents is quantitatively consistent with cholinergic action at postsynaptic muscarinic receptors of the non-M1 subtype.

Animals↗

PubMed1990

The predictive validity of teacher nominations: a five-year followup of at-risk youth.

The purpose of this study was to examine the predictive validity of a teacher nomination procedure for identifying at-risk children. Two hundred and twenty-five children were nominated by their teachers as well-adjusted (n = 75), socially withdrawn (n = 76), or socially aggressive (n = 74) during the fourth grade. Five years later, 198 of these children (88%) were located and their adjustment evaluated. Significant differences among the nominated children were found on a variety of measures, including academic grades, sociometric status, and social behavior. In addition, differences in school dropout and delinquent offenses were noted. Results are discussed in terms of the validity of teacher nomination procedures and their utility in identifying at-risk youth.

Adolescent↗

PubMed1990

Effects of histamine on dentate granule cells in vitro.

Hippocampal slices from rat brain were exposed to histamine and related substances in a perfusion chamber. Granule cells of the dentate gyrus were studied with conventional extra- and intracellular recording and a single electrode voltage clamp. Histamine caused, through activation of H(2)-receptors, a small depolarization, an increase in the number of synaptic and action potentials, a block of the long lasting (but not the early) component of spike afterhyperpolarizations and a reduction of the accommodation of action potential firing. These effects were mimicked by forskolin (suggests activation of adenylate cyclase). In voltage clamp, histamine blocked a long lasting calcium-dependent outward tail current without any reduction of inward current. Thus histamine selectively blocks the late calcium-dependent potassium current in dentate granule cells which receive histaminergic input from the posterior hypothalamus. Histamine also reduces the field excitatory postsynaptic potential evoked by perforant path stimulation. These actions allow for a powerful modulation of excitatory signals and an effective regulation of hippocampal excitability.

Action Potentials↗

PubMed1990

Two transient outward currents in histamine neurones of the rat hypothalamus in vitro.

1. The transient outward current exhibited by the histamine neurones of the tuberomammillary nucleus was studied using the single-electrode voltage clamp technique in an in vitro rat hypothalamic slice preparation. 2. The transient outward current exhibited steady-state inactivation at the resting potential. Inactivation was removed by priming hyperpolarization with a V1/2 of -85 +/- 1.2 mV, while the V1/2 for activation was -60.3 +/- 2.1 mV. 3. The decay of the transient outward current was best fitted by two exponentials with time constants of 104 +/- 36 and 568 +/- 128 ms. These two components were provisionally termed IA,f and IA,s for the fast and slowly decaying currents, respectively. 4. Removal of inactivation was time dependent; inactivation was fully removed by hyperpolarizing pulses to -110 mV of 200 ms or greater duration. Removal of inactivation of IA,f was rapid, becoming complete with hyperpolarizing pre-pulses of 50 ms or greater, while removal of inactivation of IA,s was not complete until hyperpolarizing pre-pulses were 200 ms in duration. 5. The fast decaying current IA,f was selectively blocked by 1 mM-4-aminopyridine. Tetraethylammonium chloride (10 mM) had no effect on either IA,f or IA,s. 6. The inactivation curves for IA,s, determined both by using the values obtained from the amplitude of the computed slower exponential function as well as that of the current remaining in 1 mM 4-aminopyridine, were negative to those of IA,f. Similarly derived activation curves for IA,s were positive to those of IA,f. 7. Superfusion with a nominal 0 Ca2+ medium containing 10 mM-Mg2+ did not reduce the maximal transient outward current. 8. The reversal potential of IA,s with 2.5 mM-K+ in the medium was -95 +/- 3 mV; the reversal potential of IA,f was at least 15 mV negative to that of IA,s. 9. It is concluded that histaminergic tuberomammillary neurones possess at least two transient outward currents which can be distinguished on the basis of their rates of decay, 4-aminopyridine sensitivity, voltage dependence and reversal potentials.

4-Aminopyridine↗

PubMed1989

Cholinergic activation of medial pontine reticular formation neurons in vitro.

Direct microinjection of cholinergic compounds into pontine reticular formation furnishes an excellent phenomenological model of the rapid eye movement phase of sleep (REM), but the mechanisms underlying this effect and whether they mimic the cellular events of natural REM remain unknown. Data presented here from intracellular recordings in vitro in the rat demonstrate that two-thirds of medial pontine reticular formation neurons respond to application of 0.5-1.0 microM carbachol with a depolarization characterized by a decreased conductance and a linear I/V curve. The resultant mimicry of REM cellular events by carbachol extends to membrane potential depolarization, increased cellular excitability, enhancement of PSPs from reticular stimulation, and the absence of a burst discharge pattern. The presence of these effects with tetrodotoxin and their blockade by atropine imply a direct, muscarinic cholinergic mediation. Other neurons tested responded with either a biphasic hyperpolarization-depolarization or a hyperpolarization. The hyperpolarization was associated with an increased conductance which exhibited pronounced inward rectification, an effect novel for cholinergic agonists in vertebrate CNS but described in heart cells.

Action Potentials↗

PubMed1989

Cholinergic activation of medial pontine reticular formation neurons in vitro.

In vivo microinjections of cholinergic compounds into the medial pontine reticular formation have produced some or depending on the injection site, all of the phenomena of REM, thus providing the only adequate pharmacological model of this behavioral state. The necessary anatomical substrate, a cholinergic projection to the mPRF was recently demonstrated, however the direct effect of cholinergic agonists on mPRF neurons is unknown. We have examined the effects of carbachol on mPRF neurons recorded in vitro from brainstem slices of Sprague-Dawley rats (8-10 days old). Three kinds of response to the application of carbachol (0.5-1 microM) were observed (n = 15) as follows: a depolarizing response (67%), a hyperpolarizing response (20%) and a biphasic response consisting of a hyperpolarizing response followed by a depolarizing response (13%). Under voltage clamp control, the depolarizing response was observed as an inward current resulting from a decrease in conductance which was constant over the membrane potential range of -100 to -50 mV. Reversal potential was negative to -80 mV. An increase in the excitability of neurons (as measured by responses to identical intracellularly applied depolarizing current pulses) during the depolarizing responses was due to the increase in steady state inward current. When intracellular DC current of equal amplitude but opposite polarity was applied, no increase in excitability was observed. This response was always blocked by the addition of atropine (0.5-1 microM) to the perfusate. The hyperpolarizing response was observed as an increase in outward current due to an increase in conductance with marked voltage sensitivity (over the range of -100 to -50 mV) characteristic of the anomalous rectifier. Preliminary data indicated that the hyperpolarizing response was more sensitive to pirenzepine (complete blockade at 1.0 microM) than the depolarizing response (complete blockade at 2 microM) but neither response was affected by pirenzepine concentrations of 200 nM or less. Cholinergic effects on evoked depolarizing PSPs were examined on neurons with depolarizing (n = 3) and biphasic (n = 1) responses and in all cases, the PSPs were enhanced. This enhancement was blocked by atropine. In conclusion, it is suggested that activation of two different muscarinic receptors (neither of which is the M1 receptor) on mPRF neurons results in two different responses, a decrease in a voltage-insensitive potassium conductance and an increase in the anomalous rectifier.

Acetylcholine↗

PubMed1989

The brain histamine system in vitro.

The electrophysiological properties of identified tuberomammillary histamine neurones were investigated in explant and slice preparations. The effects of histamine were studied on target neurones, mainly in the hippocampal slice. The results describe an important modulatory role of this diffusely projecting system.

Action Potentials↗

PubMed1989

Repetitive firing properties of medial pontine reticular formation neurones of the rat recorded in vitro.

1. Intracellularly recorded neurones in nucleus reticularis pontis caudalis of the medial pontine reticular formation (mPRF) in the in vitro slice preparation were analysed for repetitive firing properties in response to intracellularly applied constant-current pulses. 2. Three neuronal classes were defined by this procedure: (1) non-burst neurones, which had only a non-burst firing pattern; (2) low-threshold burst neurones, which had either a low-threshold burst pattern or a non-burst pattern; (3) high-threshold burst neurones, which had either a high-threshold burst pattern or a non-burst pattern. 3. Histological characterization of electrophysiologically identified mPRF neurones with carboxyfluorescein showed no definite morphological difference between the first two classes. There was a trend for low-threshold burst neurones to have larger somata. 4. The low-threshold burst was generated by a slow calcium-dependent low-threshold spike, revealed in the presence of tetrodotoxin. The size of the low-threshold spike and thus the number of fast action potentials in the low-threshold burst was controlled by at least five factors including: activation; inactivation; amplitude of low-threshold conductance available to be activated; delayed outward conductance; and early transient outward conductance. 5. The non-burst pattern examined in both non-burst and low-threshold burst neurones appeared to be controlled primarily by one or more calcium-dependent potassium conductances sensitive to the removal of calcium and tetraethyl-ammonium. In the presence of tetrodotoxin (TTX), the addition of antagonists to calcium-dependent potassium current revealed a slow high-threshold calcium spike which was distinguished from the low-threshold spike by its threshold, lack of inactivation (at potentials negative to -40 mV) and insensitivity to Mg2+. A long-duration after-hyperpolarization (greater than 0.5 s) was not observed in any of these cells. 6. An early transient outward rectification sensitive to 4-aminopyridine and probably mediated by A-current was apparent in low-threshold burst and non-burst neurones and affected both the low-threshold burst and non-burst firing patterns. 7. Alteration of resting membrane potential, such as occurs in vivo during the depolarization associated with desynchronized sleep, may inactivate the low-threshold spike and the transient outward conductance responsible for the distinctive responses observed from more hyperpolarized membrane potentials and produce a more homogeneous non-burst response pattern. Membrane potential effects on intrinsic conductances thus may furnish an important mechanism for changes in mPRF neuronal responsivene

Action Potentials↗

PubMed1989

Characterization of inhibition mediated by adenosine in the hippocampus of the rat in vitro.

1. Intracellular recordings with single-electrode voltage clamp were employed to study the mechanism of adenosine-elicited inhibition of CA1 neurones of the rat in vitro. 2. Adenosine elicits a steady-state outward current in association with an increase in conductance. The driving force varied with external potassium concentration as predicted by the Nernst equation for a change primarily in potassium permeability. 3. Adenosine current was blocked by high concentrations of 4-aminopyridine or barium. In the majority of neurones this current was voltage insensitive. In the remainder, the current was inwardly rectifying. The rectification was blocked by tetraethylammonium. 4. When the adenosine-elicited potassium current was blocked, slow inward currents, normally carried by calcium, were unaffected by adenosine. We conclude that this adenosine inhibition is mediated by an increase in a voltage- and calcium-insensitive potassium conductance in CA1 neurones.

4-Aminopyridine↗

PubMed1988

Endogenous adenosine inhibits hippocampal CA1 neurones: further evidence from extra- and intracellular recording.

Extracellular and intracellular recordings from CA1 pyramidal neurones of rats in vitro were used to study the effects of endogenous and exogenously applied adenosine. The adenosine receptor antagonist, caffeine, enhanced the intracellular recorded e.p.s.p.-i.p.s.p. sequence evoked by stimulation of the stratum radiatum which is antagonized by exogenous adenosine. The late, potassium dependent i.p.s.p. was not antagonized. The adenosine uptake inhibitor, nitrobenzylthioinosine (NBTI), mimicked the effects of exogenously applied adenosine. The effects of NBTI and of exogenously applied adenosine were antagonized by caffeine in the same manner. Exposure to adenosine deaminase enhanced the evoked field e.p.s.p. During this enhancement caffeines effects were significantly reduced. In low calcium high magnesium medium which abolishes synaptic activity, adenosine deaminase increased, NBTI decreased cell firing. We conclude that endogenous adenosine, release by a calcium independent mechanism, can exert an inhibitory tone on CA1 neurones in vitro. This is consistent with a role for adenosine as a mediator of negative feedback between the metabolic state and electrophysiological activity of nervous tissue.

Adenosine↗

PubMed1987

Action and location of neuropeptide tyrosine (Y) on hippocampal neurons of the rat in slice preparations.

The action of bath applied NPY (1-1,000 nM) was investigated on hippocampal slices of the rat with extra- and intracellular recording. Neuropeptide Y (NPY) at 10-1,000 nM caused a concentration-dependent, long-lasting reduction of excitatory postsynaptic potentials (EPSPs) in the hippocampal subfield CA1 and the area dentata, and an even stronger reduction of population spikes. Paired pulse experiments with low intensity, stimulation-evoked PSPs showed a marked increase in facilitation in the presence of NPY, indicating a presynaptic action. Spontaneous burst firing of CA1 pyramidal cells in low calcium, high magnesium medium was reduced, indicating a partially postsynaptic inhibitory action of NPY on their dendrites. Intracellular recording from CA1 somata during NPY administration revealed a reduction of the amplitudes of excitatory-inhibitory postsynaptic potential (EPSP-IPSP) sequences in the absence of changes in membrane potential and conductance. Accommodation of firing during long depolarizing pulses and afterhyperpolarizations were unchanged. The innervation pattern of NPY immunoreactive fibers in the same regions was studied in slices adjacent to the ones used for electrophysiology by using antisera against NPY and light and electron microscopy. There is a dense innervation of CA1 by NPY-immunoreactive axons and terminals, particularly in the stratum moleculare. NPY-immunoreactive neurons are present in the stratum oriens and pyramidale. The NPY labeled axons of the stratum moleculare participate in numerous synaptic contacts with the smaller dendritic elements in this layer, many of which belong to pyramidal neurons. These observations provide evidence for a dendritic NPY-immunoreactive innervation of CA1 neurons, which is in keeping with the electrophysiological effects of NPY on pyramidal neurons.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1986

A low threshold calcium spike mediates firing pattern alterations in pontine reticular neurons.

Intracellular electrical recordings in an in vitro slice preparation of the brainstem medial pontine reticular formation, a region thought to be important in mediation of desynchronized sleep phenomena, demonstrate a population of neurons that have a calcium-dependent, low threshold spike. This low threshold spike was inactivated at relatively depolarized membrane potential levels and, when this spike was deinactivated, it induced a burst of action potentials. The membrane potential dependence of the spike may underlie changes in action potential firing patterns associated with behavioral state change because the baseline membrane potential in neurons of the medial pontine reticular population depolarizes during passage from waking and slow wave sleep to desynchronized sleep, which is characterized by the absence of burst firing.

Action Potentials↗

PubMed1986

Effects of histamine on hippocampal pyramidal cells of the rat in vitro.

The actions of bath applied histamine on CA1 pyramidal cells were investigated in hippocampal slices of the rat. Histamine caused a) a slight depolarization but no significant change in resting membrane conductance; b) an abbreviation of long afterhyperpolarizations after single action potentials, bursts of action potentials or TTX resistant spikes; c) a loss of accommodation of firing. In the presence of TEA or barium, histamine prolonged and increased the size and number of the slow TTX resistant spikes. A depolarizing plateau which follows such spikes was also increased by histamine, but the population spike was increased. The frequency of spontaneous chloride dependent potentials, which reflect interneurone firing, was also increased. These effects considerably outlasted histamine application and were mimicked by the H2-agonist impromidine but not the H1-agonist thiazolethylamine, and blocked by the H2-antagonists cimetidine and metiamide but not the H1-antagonists mepyramine or the beta-antagonist propranolol. It is concluded that histamine, by activating H2-receptors, antagonizes a calcium mediated potassium conductance in hippocampal pyramidal cells without affecting calcium current. By this mechanism histaminergic afferent fibres could effectively regulate cortical responsiveness by selectively potentiating large excitatory inputs of target neurones.

Animals↗

PubMed1986

A simple in vitro method to study the trigeminal ganglion.

This report describes a simple in vitro method to harvest and study the electrophysiological properties of the trigeminal ganglion of the rat. In suction electrode recordings from the proximal nerve end, two distinct peaks are identified in the compound action potential evoked by electrical stimulation of the distal nerve. gamma-Aminobutyric acid (GABA), added to the perfusion fluid, caused a positive shift in the DC potential (depolarization) and a partially selective decrease in the amplitude of the slow-conduction peak. Reversible antagonism of the GABA effect by picrotoxin and bicuculline suggests that both responses are receptor mediated. There was no response to carbamazepine or L-baclofen, suggesting that ganglionic polarization does not play a major role in the action of these drugs in trigeminal neuralgia.

Action Potentials↗

PubMed1985

Stereoselectivity of L-baclofen in hippocampal slices of the rat.

Extra- and intracellular recording from hippocampal slices of the rat revealed the following effects when baclofen (BF) (0.1-10 microM) was added to the perfusion fluid: a block of synaptic potentials evoked by stimulation of stratum radiatum; a direct hyperpolarization and a conductance increase (for potassium ions) of CA1 pyramidal cells. All this activity was found in the L- none in the D-enantiomer. D-BF did not antagonize the action of L-BF.

Animals↗

PubMed1985

Long-term potentiation and 4-aminopyridine.

Long-term potentiation (LTP) of excitatory postsynaptic potentials (epsp's) was investigated with extracellular field potential recording in hippocampal slices from rats. In the presence of 100 microM 4-aminopyridine (4-AP) the probability of eliciting LTP was unchanged or increased; the extent of potentiation was not significantly different from normal. During LTP saturation, 4-AP further enhanced the epsp. These data are inconsistent with an involvement of A-current reduction in LTP.

4-Aminopyridine↗

PubMed1985

Effects of caffeine on hippocampal pyramidal cells in vitro.

The effects of caffeine on the electrophysiological properties of CA1 pyramidal neurones were investigated in the rat hippocampal slice preparation in vitro. A concentration-dependent increase in both the extracellularly recorded excitatory postsynaptic potential (e.p.s.p.) and the population spike resulting from stimulation of the stratum radiatum could be evoked by caffeine with a threshold concentration of 10 microM. Intracellular recordings demonstrate a caffeine-evoked decrease in resting membrane potential, an increase in input resistance, a reduction of the long afterhyperpolarization (a.h.p.) and a decrease in accommodation. The interaction between caffeine and adenosine was investigated on the extracellularly recorded e.p.s.p. The maximal response evoked by caffeine was increased in the presence of adenosine and the adenosine concentration-response curve was shifted to the right in a parallel fashion in the presence of caffeine. It is suggested that the effects of caffeine on hippocampal neurones may be mediated by a decrease of one or more potassium conductance(s), and that adenosine and caffeine may compete for the same electrophysiologically active receptor site on these cells.

Adenosine↗

PubMed1985

Adenosine actions on CA1 pyramidal neurones in rat hippocampal slices.

Intracellular recordings with a bridge amplifier of CA1 pyramidal neurones in vitro were employed to study the mechanisms of action of exogenously applied adenosine in the hippocampal slice preparation of the rat. Adenosine enhanced the calcium-dependent, long-duration after-hyperpolarization (a.h.p.) at least in part by a reduction in the rate of decay of the a.h.p. Both the reduced rate of decay and that of the control can be described with a single exponential. Antagonism of the calcium-dependent potassium current (and as a result, the a.h.p.) by bath application of CdCl2 or intracellular injection of EGTA (ethyleneglycolbis-(beta-aminoethyl ether)N,N'-tetraacetic acid) did not reduce the adenosine-evoked hyperpolarization or decrease in input resistance. Similarly, TEA (tetraethylammonium), which antagonizes both the voltage- and calcium-sensitive, delayed, outward rectification, had no effect on the adenosine-evoked changes in resting membrane properties. Adenosine did not affect the early, transient, outward rectification. During exposure to 4-aminopyridine (4-AP) in concentrations sufficient to antagonize this early rectification, the changes in resting membrane properties evoked by adenosine were unaffected. We conclude that the enhancement of the a.h.p. and accommodation by adenosine may be mediated by a change in the regulation of intracellular calcium. However, the mechanism responsible for the hyperpolarization and decrease in input resistance evoked by adenosine is both calcium and voltage insensitive. Thus, it appears distinct from that mediating the enhancement of the a.h.p. and accommodation.

Action Potentials↗