Loss of cholinergic neurons in the nucleus basalis induces neocortical electroencephalographic and passive avoidance deficits.

The present experiments were designed to examine the hypothesis that the degeneration of cholinergic nucleus basalis is related to the cognitive and neurophysiological deficits found in old age. Aged (26 months) rats were impaired both in the acquisition of spatial (water-maze) task and retention of passive avoidance task. During aging, neocortical electroencephalographic fast activity was decreased and high-voltage spindles increased. Loss of choline acetyltransferase-positive neurons correlated with the high-voltage spindle incidence and passive avoidance retention deficit. Unilateral ibotenate nucleus basalis lesioning decreased choline acetyltransferase activity in the cortex and produced a large nonspecific subcortical cell loss in young rats. Ibotenate-lesioned rats were impaired in spatial learning and passive avoidance retention in young rats. Quisqualic acid produced a greater decrease in cortical choline acetyltransferase activity and smaller nonspecific subcortical cell loss than ibotenate lesioning. Spatial learning was not impaired, but passive avoidance performance was disrupted. Slow waves and high-voltage spindles were increased and beta activity decreased on the side of either quisqualate or ibotenate nucleus basalis lesioning. These results demonstrate that age-related neurophysiological and cognitive deficits result partially from the loss of cholinergic neurons in the nucleus basalis and that quisqualic acid nucleus basalis-lesioning in young rats may be used as a pharmacological model of the age-related cholinergic neuron loss.     

Low-threshold calcium electrogenesis in neocortical neurons

In slices of parietal neocortex, evidence was obtained for the existence of a low-threshold Ca2+ conductance in most neurons. This conductance became apparent when resting membrane potential was held below -60 mV by continuously injected, depolarizing current. Under these conditions, brief hyperpolarizing pulses were followed by generation of tetrodotoxin (TTX)-resistant, Mn2+-sensitive, low-threshold spikes. The results suggest that in neocortex, as in many subcortical structures, low-threshold Ca current may be responsible for burst generation in some neurons.     

Cerebral neocortical neurons in the aged rat: spontaneous activity, properties of pyramidal tract neurons and effect of acetylcholine and cholinergic drugs

The properties of cortical cerebral neurons have been studied and compared in 2, 22 and 26 month-old Sprague-Dawley rats, using electrophysiological techniques. The mean spontaneous activity of the neurons in old animals (unidentified as well as pyramidal tract neurons) was not different from that of young adult rats. In contrast the mean latency of the antidromic response of pyramidal tract neurons to pyramidal tract stimulation was significantly longer in 26 month-old animals. No difference was observed in the effects of the excitatory amino acid glutamate applied by iontophoresis. The percentage of cortical neurons excited by the iontophoretic application of acetylcholine was similar in young and old animals. Neither the laminar distribution, nor the individual sensitivity of these neurons to acetylcholine were found to be modified. The pharmacological properties of the acetylcholine-induced excitations were unchanged, exhibiting muscarinic as well as nicotinic properties. These results are consistent with the suggestion that the impairment of the cholinergic system with aging is for a large part presynaptic. They also emphasize the fact that several physiological and pharmacological properties of the cerebral cortical neurons show little change with age in Sprague-Dawley rats.     

Electrophysiological properties of neocortical neurons in vitro

1. Intracellular recordings were obtained from neurons of the guinea pig sensorimotor cortical slice maintained in vitro. Under control recording conditions input resistances, time constants, and spiking characteristics of slice neurons were well within the ranges reported by other investigators for neocortical neurons in situ. However, resting potentials (mean of -75 mV) and spike amplitudes (mean of 93.5 mV) were 10-25 mV greater than has been observed in intact preparations. 2. Current-voltage relationships obtained under current clamp revealed a spectrum of membrane-rectifying properties at potentials that were subthreshold for spike generation. Ionic and pharmacologic analyses suggest that subthreshold membrane behavior is dominated by voltage-sensitive, very slowly inactivating conductances to K+ and Na+. 3. Action potentials were predominantly Na+ dependent under normal conditions but when outward K+ currents were reduced pharmacologically, it was possible, in most cells, to evoke a non-Na+-dependent, tetrodotoxin-(TTX) insensitive spike, which was followed by a prominent depolarizing after-potential. Both of these events were blocked by the Ca2+ current antagonists, Co2+ and Mn2+. 4. A small population of neurons generated intrinsic, all-or-none burst potentials when depolarized with current pulses or by synaptic activation. These cells were located at a narrow range of depths comprising layer IV and the more superficial parts of layer V. 5. Spontaneous excitatory synaptic potentials appeared in all neurons. Spontaneous inhibitory events were visible in only about 10% of the cells, and in those cases apparently reversed polarity at a level slightly positive to resting potential. Stimulation of the surface of the slice at low intensities evoked robust and usually concurrent excitatory and inhibitory synaptic potentials. Unitary inhibitory postsynaptic potentials (IPSPs) reversed at levels positive to rest. Stronger stimulation produced a labile, long-duration, hyperpolarizing IPSP with a reversal potential 15-20 mV negative to the resting level. 6. Neocortical neurons in vitro retain the basic membrane and synaptic properties ascribed to them in situ. However, the array of passive and active membrane behavior observed in the slice suggests that cortical neurons may be differentiated by specific functional properties as well as by their extensive morphological diversity.     

GABA-induced calcium signaling in cultured enteric neurons is reinforced by activation of cholinergic pathways

GABA is an important inhibitory transmitter in the CNS. In the enteric nervous system, however, both excitatory and inhibitory actions have been reported. Here, we investigated the effects of GABA on the intracellular Ca2+ concentration of guinea-pig myenteric neurons (at 35 degrees C) using Fura-2-AM. Neurons were identified by 75 mM K+ depolarization (5 s), which evoked a transient intracellular Ca2+ concentration increase. GABA (10 s) induced a dose dependent (5 nM-1 microM) transient intracellular Ca2+ concentration rise in the majority of neurons (500 nM GABA: 251+/-17 nM, n=232/289). Interestingly, the response to 5 microM GABA (n=18) lasted several minutes and did not fully recover. GABA response amplitudes were significantly (P<0.001) reduced by GABAA and GABAB receptor antagonists (10 microM) bicuculline and phaclofen. The GABAA agonist isoguvacine (10 microM) and GABAB agonist baclofen (10 microM) induced similar responses as 50 nM GABA, while the GABAC agonist cis-4-aminocrotonic acid (CACA) (10 microM) only elicited small responses in a minority of neurons. Removal of extracellular Ca2+ abolished all responses while depletion of intracellular Ca2+ stores by thapsigargin (5 microM) did not alter the responses to 500 nM GABA (n=13), but reduction of Ca2+ influx through voltage-dependent Ca2+ channels did. The nicotinic antagonist hexamethonium (100 microM) also reduced GABA responses by almost 70% suggesting that GABA stimulates cholinergic pathways, while the purinergic receptor blocker pyridoxal-phosphate-6-azophenyl-2',4'-disulfonic acid (PPADS) and the 5-HT3 receptor blocker ondansetron only had minor effects. Conclusion: GABA elicits transient intracellular Ca2+ concentration responses in the majority of myenteric neurons through activation of GABAA and GABAB receptors and much of the response can be attributed to facilitation of ACh release. Thus GABA may act mainly as a modulator that sets the state of excitability of the enteric nerve network. A concentration of 5 microM GABA, although frequently used in pharmacological experiments, seems to cause a detrimental response reminiscent of the neurotoxic effects glutamate has in the CNS.     

Two patterns of firing in human neocortical neurons

Intracellular recordings were made in slices of human neocortex removed for surgical treatment of epilepsy. In response to prolonged suprathreshold current injection, regular spiking neurons (67.5% of sample) responded by repetitive firing throughout the stimulus from all membrane potentials. Bursting neurons (32.5% of sample) responded with a burst of 2–3 spikes which rode upon a voltage-dependent slow depolarization. Bursting behavior was only observed from membrane potentials more negative than −65 mV.     

Electrophysiological characteristics of neurons in neocortical expiant cultures

Summary We examined the electrophysiological and morphological properties of neocortical neurons maintained in expiant cultures prepared from the parietal cortex of newborn Sprague-Dawley rats. After 3–6 weeks in vitro, cultures showed regional differences in cellular density reminiscent of cortical layering, and an abundance of axonal processes. Pyramidal-shaped neurons with spinous dendrites were the dominant elements revealed by Lucifer yellow injections. Intracellular recordings revealed that many electrophysiological properties of neurons in the explants resembled those of neocortical neurons in vivo and in slice preparations. In response to depolarizing current injection, neurons in the expiants showed the same three patterns of repetitive firing described in neocortical slices, as well as a similar array of responses. Spontaneous synaptic potentials were recorded from all neurons and complex PSPs were evoked in response to focal extracellular stimulation. GABAa receptors mediated a significant component of the evoked responses. Fifteen of sixty neurons generated action potentials that arose spontaneously from resting potentials. Neurons in many slices generated large, prolonged depolarizing potentials that reflected coordinated synaptic activity within the expiants. These results underscore the usefulness of the neocortical explant as a valuable model for studying aspects of the behavior of circuits of cortical neurons.     

Fractional differentiation by neocortical pyramidal neurons

Neural systems adapt to changes in stimulus statistics. However, it is not known how stimuli with complex temporal dynamics drive the dynamics of adaptation and the resulting firing rate. For single neurons, it has often been assumed that adaptation has a single time scale. We found that single rat neocortical pyramidal neurons adapt with a time scale that depends on the time scale of changes in stimulus statistics. This multiple time scale adaptation is consistent with fractional order differentiation, such that the neuron's firing rate is a fractional derivative of slowly varying stimulus parameters. Biophysically, even though neuronal fractional differentiation effectively yields adaptation with many time scales, we found that its implementation required only a few properly balanced known adaptive mechanisms. Fractional differentiation provides single neurons with a fundamental and general computation that can contribute to efficient information processing, stimulus anticipation and frequency-independent phase shifts of oscillatory neuronal firing.     

Activity of beta-kainic acid on neocortical neurons in vivo and hippocampal neurons in vitro

The beta-isomer of kainic acid has been reported to have anticonvulsant activity with a profile suggesting amino acid antagonism. The present study, on neurons in the cerebral cortex of anaesthetised rats or in hippocampal slices, shows that beta-kainate does not antagonise any of the major agonists at amino acid receptors: N-methylaspartate, quisqualic acid or alpha-kainic acid. beta-Kainate does have an excitant action of its own which can be partially reduced by 2-amino-5-phosphonovaleric acid but is unaffected by gamma-D-glutamylglycine. It is blocked completely by kynurenic acid.     

Voltage-activated sodium channels amplify inhibition in neocortical pyramidal neurons

Inhibitory postsynaptic potentials (IPSPs) in neocortical pyramidal neurons are increased in duration and amplitude at depolarized membrane potentials. This effect was not due to changes in the time course of the underlying synaptic current. The role of postsynaptic voltage-activated channels was investigated by mimicking the voltage change that occurs during an IPSP with current injections. The peak and integral of these 'simulated' IPSPs increased during depolarization of the membrane potential in a tetrodotoxin-sensitive manner. This amplification presumably occurs as the hyperpolarization associated with IPSPs turns off sodium channels that are tonically active at depolarized membrane potentials. IPSP amplification increased the ability of IPSPs to inhibit action potential firing and promoted IPSP-induced action potential synchronization.     

Regulation of cholinergic phenotype in developing neurons

Specification of neurotransmitter phenotype is critical for neural circuit development and is influenced by intrinsic and extrinsic factors. Recent findings in rat hypothalamus in vitro suggest the role of neurotransmitter glutamate in the regulation of cholinergic phenotype. Here we extended our previous studies on the mechanisms of glutamate-dependent regulation of cholinergic phenotypic properties in hypothalamic neurons. Using immunocytochemistry, electrophysiology, and calcium imaging, we demonstrate that hypothalamic expression of choline acetyltransferase (the cholinergic marker) and responsiveness of neurons to acetylcholine (ACh) receptor agonists increase during chronic administration of an N-methyl-D-aspartate receptor (NMDAR) blocker, MK-801, in developing rats in vivo and genetic and pharmacological inactivation of NMDARs in mouse and rat developing neuronal cultures. In hypothalamic cultures, an inactivation of NMDA receptors also induces ACh-dependent synaptic activity, as do inactivations of PKA, ERK/MAPK, CREB, and NF-kappaB, which are known to be regulated by NMDA receptors. Interestingly, the increase in cholinergic properties in developing neurons that is induced by NMDAR blockade is prevented by the blockade of ACh receptors, suggesting that function of ACh receptor is required for the cholinergic up-regulation. Using dual recording of monosynaptic excitatory postsynaptic currents, we further demonstrate that chronic inactivation of ionotropic glutamate receptors induces the cholinergic phenotype in a subset of glutamatergic neurons. The phenotypic switch is partial as ACh and glutamate are coreleased. The results suggest that developing neurons may not only coexpress multiple transmitter phenotypes, but can also change the phenotypes following changes in signaling in neuronal circuits.     

Dynamics of population rate codes in ensembles of neocortical neurons

Information processing in neocortex can be very fast, indicating that neuronal ensembles faithfully transmit rapidly changing signals to each other. Apart from signal-to-noise issues, population codes are fundamentally constrained by the neuronal dynamics. In particular, the biophysical properties of individual neurons and collective phenomena may substantially limit the speed at which a graded signal can be represented by the activity of an ensemble. These implications of the neuronal dynamics are rarely studied experimentally. Here, we combine theoretical analysis and whole cell recordings to show that encoding signals in the variance of uncorrelated synaptic inputs to a neocortical ensemble enables faithful transmission of graded signals with high temporal resolution. In contrast, the encoding of signals in the mean current is subject to low-pass filtering.