Network recruitment to coherent oscillations in a hippocampal computer model

Coherent neural oscillations represent transient synchronization of local neuronal populations in both normal and pathological brain activity. These oscillations occur at or above gamma frequencies (>30 Hz) and often are propagated to neighboring tissue under circumstances that are both normal and abnormal, such as gamma binding or seizures. The mechanisms that generate and propagate these oscillations are poorly understood. In the present study we demonstrate, via a detailed computational model, a mechanism whereby physiological noise and coupling initiate oscillations and then recruit neighboring tissue, in a manner well described by a combination of stochastic resonance and coherence resonance. We develop a novel statistical method to quantify recruitment using several measures of network synchrony. This measurement demonstrates that oscillations spread via preexisting network connections such as interneuronal connections, recurrent synapses, and gap junctions, provided that neighboring cells also receive sufficient inputs in the form of random synaptic noise. "Epileptic" high-frequency oscillations (HFOs), produced by pathologies such as increased synaptic activity and recurrent connections, were superior at recruiting neighboring tissue. "Normal" HFOs, associated with fast firing of inhibitory cells and sparse pyramidal cell firing, tended to suppress surrounding cells and showed very limited ability to recruit. These findings point to synaptic noise and physiological coupling as important targets for understanding the generation and propagation of both normal and pathological HFOs, suggesting potential new diagnostic and therapeutic approaches to human disorders such as epilepsy.     

Fast network oscillations induced by potassium transients in the rat hippocampus in vitro

Brief pressure ejection of solutions containing potassium, caesium or rubidium ions into stratum radiatum of the CA1 or CA3 regions of the hippocampal slice evoked a fast network oscillation. The activity evoked lasted ∼3–25 s with the predominant frequency component being in the gamma frequency range (30–80 Hz), although beta frequency (15–30 Hz) and ultrafast (> 80 Hz) components could also be seen. The gamma frequency component of the oscillation remained constant, even when large changes in power occurred, and was synchronous across the CA1 region. Measurements with potassium ion-sensitive electrodes revealed that the network oscillation was accompanied by increases (0.5 to 2.0 m m ) in the extracellular potassium concentration [K⁺]_o. Bath application of the N -methyl- d -aspartate (NMDA) receptor antagonists d (–)-2-amino-5-phosphonopentanoic acid ( d -AP5; 50 μM) had no significant effect but the α-amino-3-hydroxy-5-methyl-4-isooxazolepropionic acid (AMPA)/kainate receptor antagonist 2,3,-dioxo-6-nitro-1,2,3,4-tetrahydrobenzo[f]quinoxaline-7-sulphonamide disodium (NBQX; 20 μM) caused a significant reduction (86.7 ± 4.5 %) in the power in the gamma frequency range. Residual rhythmic activity, presumably arising in the interneuronal network, was blocked by the GABA_A receptor antagonist bicuculline. The putative gap junction blocker octanol caused a decrease in the power of the gamma frequency component of 75.5 ± 5.6 %, while carbenoxolone produced a reduction of only 14 ± 42 %. These experiments demonstrate that a modest increase in exogenous [K⁺]_o in the hippocampus in vitro is sufficient to evoke a fast network oscillation, which is an emergent property of the synaptically and electrically interconnected neuronal network.     

Region-specific modulation of electrically induced synchronous oscillations in the rat hippocampus and cerebral cortex

Strong tetanization induces synchronous membrane potential oscillations (seizure-like afterdischarge) in mature pyramidal cells of the hippocampal CA1 region. To investigate whether local networks in other brain regions can generate such an afterdischarge independently, we studied the inducibility of afterdischarge in individual 'isolated slices' of the rat hippocampal CA1 and CA3 regions, dentate gyrus (DG), entorhinal cortex (EC), and temporal cortex (TC) using intracellular and extracellular recordings. The strong tetanization constantly induced afterdischarges in the CA1 and CA3 pyramidal cells as well as in the EC and TC superficial principal cells. However, parameters of the afterdischarges, such as the frequency and duration of afterdischarges, varied among the regions. A mixture of N-methyl-D-aspartate (NMDA) and non-NMDA receptor antagonists or a GABA(A) receptor antagonist completely blocked the afterdischarges. Local GABA application during the afterdischarge elicited depolarization, rather than hyperpolarization. Moreover, reversal potentials of the afterdischarge were around -40 mV. In contrast, the tetanization resulted in occasional afterdischarge-like activities in DG slices, which were blocked by the non-NMDA or GABA(A) receptor antagonist. These findings suggest that the afterdischarges mediated through the excitatory GABAergic and glutamatergic transmissions might be common to, but be modulated differently by individual local networks in the hippocampus and cortex.     

Generation of slow network oscillations in the developing rat hippocampus after blockade of glutamate uptake

Cell-surface glutamate transporters are essential for the proper function of early cortical networks because their dysfunction induces seizures in the newborn rat in vivo. We have now analyzed the consequences of their inhibition by DL-TBOA on the activity of the developing CA1 rat hippocampal network in vitro. DL-TBOA generated a pattern of recurrent depolarization with an onset and decay of several seconds' duration in interneurons and pyramidal cells. These slow network oscillations (SNOs) were mostly mediated by gamma-aminobutyric acid (GABA) in pyramidal cells and by GABA and N-methyl-D-aspartate (NMDA) receptors in interneurons. However, in both cell types SNOs were blocked by NMDA receptor antagonists, suggesting that their generation requires a glutamatergic drive. Moreover, in interneurons, SNOs were still generated after the blockade of NMDA-mediated synaptic currents with MK-801, suggesting that SNOs are expressed by the activation of extrasynaptic NMDA receptors. Long-lasting bath application of glutamate or NMDA failed to induce SNOs, indicating that they are generated by periodic but not sustained activation of NMDA receptors. In addition, SNOs were observed in interneurons recorded in slices with or without the strata pyramidale and oriens, suggesting that the glutamatergic drive may originate from the radiatum and pyramidale strata. We propose that in the absence of an efficient transport of glutamate, the transmitter diffuses in the extracellular space to activate extrasynaptic NMDA receptors preferentially present on interneurons that in turn activate other interneurons and pyramidal cells. This periodic neuronal coactivation may contribute to the generation of seizures when glutamate transport dysfunction is present.     

Treatment of the damaged rat hippocampus with a locally applied electric field

Summary Previous studies have indicated that axons may be directed to regenerate toward the cathodal source of a locally applied D.C. electric field. In the present studies, similar galvanotropic effects were tested after partial denervation of the rat hippocampus. Following unilateral fimbrial lesions, 1.5 A of direct current was applied locally to the tissue for 4 weeks. In the experimental group, the distal (temporal) portion of the hippocampus was located near the cathodal electrode, while the proximal (nasal) portion was located near the anode. In two control groups, either the current was reversed, or there was not current delivered to the tissue. Spontaneous alternation behavior 4 weeks post-operatively in the cathode distal rats provided results comparable to those in unoperated (normal) animals, whereas the control animals exhibited statistically significantly greater deficits in memory. The ability to learn the location of a submerged platform in a water tank was evident in unoperated and cathode distal animals, but not in either of the control groups. Hippocampal acetylcholinesterase activity in cathode distal animals was lower than in unoperated animals, but statistically significantly greater than in the other control animals. Results are consistent with the contention that locally applied weak direct current can modulate long term sequellae of hippocampal injury.     

Nonlinear cell response to strong electric fields

The response of living cells to externally applied electric fields is of widespread interest. In particular, the intensification of electric fields across cell membranes is believed to be responsible, through membrane rupture and reversible membrane breakdown processes, for certain types of tissue damage in electrical trauma cases which cannot be attributed to Joule heating. Large elongated cells such as skeletal muscle fibres are particularly vulnerable to such damage. Previous theoretical studies of field intensification across cell membranes in such cells have assumed the membrane current to be linear in the applied field (Ohmic membrane conductivity) and were limited to sinusoidal applied fields. In this paper, we investigate a simple model of a long cylindrical cell, corresponding to nerve or skeletal muscle cells. Employing the electroquasistatic approximation, a system of coupled first-order differential equations for the membrane electric field is derived which incorporates arbitrary time dependence in the external field and nonlinear membrane response (non-Ohmic conductivity). The behaviour of this model is investigated for a variety of applied fields in both the linear and highly nonlinear regimes. We find that peak membrane fields predicted by the nonlinear model are approximately twice as intense, for low-frequency electrical trauma conditions, as those of the linear theory.     

Effects of uniform extracellular DC electric fields on excitability in rat hippocampal slices in vitro

The effects of uniform steady state (DC) extracellular electric fields on neuronal excitability were characterized in rat hippocampal slices using field, intracellular and voltage-sensitive dye recordings. Small electric fields (<|40| mV mm⁻¹), applied parallel to the somato-dendritic axis, induced polarization of CA1 pyramidal cells; the relationship between applied field and induced polarization was linear (0.12 ± 0.05 mV per mV mm⁻¹ average sensitivity at the soma). The peak amplitude and time constant (15–70 ms) of membrane polarization varied along the axis of neurons with the maximal polarization observed at the tips of basal and apical dendrites. The polarization was biphasic in the mid-apical dendrites; there was a time-dependent shift in the polarity reversal site. DC fields altered the thresholds of action potentials evoked by orthodromic stimulation, and shifted their initiation site along the apical dendrites. Large electric fields could trigger neuronal firing and epileptiform activity, and induce long-term (>1 s) changes in neuronal excitability. Electric fields perpendicular to the apical–dendritic axis did not induce somatic polarization, but did modulate orthodromic responses, indicating an effect on afferents. These results demonstrate that DC fields can modulate neuronal excitability in a time-dependent manner, with no clear threshold, as a result of interactions between neuronal compartments, the non-linear properties of the cell membrane, and effects on afferents.     

Age-dependent reduction of gamma oscillations in the mouse hippocampus in vitro

Normal brain ageing is associated with a decline in hippocampal memory functions. Neuronal oscillations in the gamma frequency band have been implicated in various cognitive tasks. In this study we test the effect of normal brain ageing on gamma oscillations in the mouse hippocampus in vitro. gamma Oscillations were evoked by either 10 microM carbachol or 100 nM kainate in ventral hippocampus slices from young (>5 month) and aged (>22 month) C57Bl/J6 mice. In slices from young mice carbachol-induced gamma oscillations were more regular and more coherent than those induced by kainate. Compared with young, the power in the 20-80 Hz frequency range in area CA3 of slices from aged mice was reduced to 14% for kainate-induced oscillations and to 7% for carbachol-induced oscillations, whereas waveform, dominant frequency and coherence of the oscillation were unchanged. Local network properties were assessed by paired-pulse stimulation of Schaffer collateral/commissural fibers. The excitatory synaptic response in stratum radiatum of CA3 was reduced, in correlation with the antidromic population spike, but functional inhibition in CA3 and CA1 was unaffected. Changes in local network properties could not explain the reduced gamma oscillation strength. Since oscillations driven by two different pathways are similarly affected with age, an age-dependent effect on tonic depolarizing drive of principal cells is unlikely to explain the current results. Other mechanisms, including a change with age in the use-dependent modulation of synaptic strength, should account for the impaired gamma oscillations in the aged hippocampus that may contribute to age-dependent memory impairment.     

Fetal exposure to low frequency electric and magnetic fields

To investigate the interaction of low frequency electric and magnetic fields with pregnant women and in particular with the fetus, an anatomical voxel model of an 89 kg woman at week 30 of pregnancy was developed. Intracorporal electric current density distributions due to exposure to homogeneous 50 Hz electric and magnetic fields were calculated and results were compared with basic restrictions recommended by ICNIRP guidelines. It could be shown that the basic restriction is met within the central nervous system (CNS) of the mother at exposure to reference level of either electric or magnetic fields. However, within the fetus the basic restriction is considerably exceeded. Revision of reference levels might be necessary.     

Parvalbumin-deficiency facilitates repetitive IPSCs and gamma oscillations in the hippocampus

In the hippocampus, the calcium-binding protein parvalbumin (PV) is expressed in interneurons that innervate perisomatic regions. PV in GABAergic synaptic terminals was proposed to limit repetitive GABA release by buffering of "residual calcium." We assessed the role of presynaptic PV in Ca(2+)-dependent GABA release in the hippocampus of PV-deficient (PV-/-) mice and wild-type (PV+/+) littermates. Pharmacologically isolated inhibitory postsynaptic currents (IPSCs) were evoked by low-intensity stimulation of the stratum pyramidale and recorded from voltage-clamped CA1 pyramidal neurons. The amplitude and decay time constant of single IPSCs were similar for both genotypes. Under our experimental conditions of reduced release probability and minimal presynaptic suppression, paired-pulse facilitation of IPSCs occurred at intervals from 2 to 50 ms, irrespective of the presence of PV. The facilitation of IPSCs induced by trains of 10 stimuli at frequencies >20 Hz was enhanced in cells from PV-/- mice, the largest difference between PV-/- and PV+/+ animals (220%) being observed at 33 Hz. The effect of IPSC facilitation at sustained gamma frequencies was assessed on kainate-induced rhythmic IPSC-paced neuronal oscillations at gamma frequencies, recorded with dual field potential recordings in area CA3. The maximum power of the oscillation was 138 microV(2) at 36 Hz in slices from PV+/+ mice and was trebled in slices from PV-/- mice. PV deficiency caused a similar increase in gamma power under conditions used to study IPSC facilitation and can be explained by an increased facilitation of GABA release at sustained high frequencies. The dominant frequency and coherence were not affected by PV deficiency. These observations suggest that PV deficiency, due to an increased short-term facilitation of GABA release, enhances inhibition by high-frequency burst-firing PV-expressing interneurons and may affect the higher cognitive functions associated with gamma oscillations.     

Acetylcholine potentiates responses to N-methyl-D-aspartate in the rat hippocampus

The effect of acetylcholine (ACh) on intracellular responses to ionophoretic application of N-methyl-D-aspartate (NMDA) was examined in rat hippocampal slice. Recordings were obtained from CA1 neurons under current- and voltage-clamp conditions. Drugs were applied topically by ionophoretic and microdrop techniques. ACh produced an atropine-sensitive potentiation of responses to NMDA. The effect of ACh on NMDA receptor-mediated responses was independent of changes in voltage or potassium conductances caused by ACh. ACh also potentiated responses to L-glutamate but not to kainate or quisqualate. This effect was blocked by DL-2-amino-5-phosphonovalerate (2-APV), an NMDA receptor antagonist. We conclude that ACh, acting on muscarinic receptors, potentiates selectively, the NMDA subclass of L-glutamate receptor.     

Influence of extremely low frequency magnetic fields on Ca signaling and NMDA receptor functions in rat hippocampus

Extremely low frequency (ELF < 300 Hz) electromagnetic fields affect several neuronal activities including memory. Because ELF magnetic fields cause altered Ca²⁺ homeostasis in neural tissues, we examined their influence on Ca²⁺ signaling enzymes in hippocampus and related them with NMDA receptor functions. Hippocampal regions were obtained from brains of 21-day-old rats that were exposed for 90 days to 50 Hz magnetic fields at 50 and 100 μT intensities. In comparison to controls, ELF exposure caused increased intracellular Ca²⁺ levels concomitant with increased activities of Ca²⁺-dependent protein kinase C (PKC), cAMP-dependent protein kinase and calcineurin as well as decreased activity of Ca²⁺–calmodulin-dependent protein kinase in hippocampal regions. Simultaneous ligand-binding studies revealed decreased binding to N-methyl-d-aspartic acid (NMDA) receptors. The combined results suggest that perturbed neuronal functions caused by ELF exposure may involve altered Ca²⁺ signaling events contributing to aberrant NMDA receptor activities.     

NMDAR antagonist action in thalamus imposes delta oscillations on the hippocampus

Work on schizophrenia demonstrates the involvement of the hippocampus in the disease and points specifically to hyperactivity of CA1. Many symptoms of schizophrenia can be mimicked by N-methyl-d-aspartate receptor (NMDAR) antagonist; notably, delta frequency oscillations in the awake state are enhanced in schizophrenia, an abnormality that can be mimicked by NMDAR antagonist action in the thalamus. Given that CA1 receives input from the nucleus reuniens of the thalamus, we sought to determine whether an NMDAR antagonist in the thalamus can affect hippocampal processes. We found that a systemic NMDAR antagonist (ketamine; 50 mg/kg) increased the firing rate of cells in the reuniens and CA1 in awake rats. Furthermore, ketamine increased the power of delta oscillations in both structures. The thalamic origin of the change in hippocampal properties was demonstrated in three ways: 1) oscillations in the two structures were coherent; 2) the hippocampal changes induced by systematic ketamine were reduced by thalamic injection of muscimol; and 3) the hippocampal changes could be induced by local injection of ketamine into the thalamus. Lower doses of ketamine (20 mg/kg) did not evoke delta oscillations but did increase hippocampal gamma power, an effect not dependent on the thalamus. There are thus at least two mechanisms for ketamine action on the hippocampus: a low-dose mechanism that affects gamma through a nonthalamic mechanism and a high-dose mechanism that increases CA1 activity and delta oscillations as a result of input from the thalamus. Both mechanisms may be important in producing symptoms of schizophrenia.     

Monopole patch antenna for in vivo exposure to nanosecond pulsed electric fields

To explore the promising therapeutic applications of short nanosecond electric pulses, in vitro and in vivo experiments are highly required. In this paper, an exposure system based on monopole patch antenna is reported to perform in vivo experiments on newborn mice with both monopolar and bipolar nanosecond signals. Analytical design and numerical simulations of the antenna in air were carried out as well as experimental characterizations in term of scattering parameter (S ₁₁) and spatial electric field distribution. Numerical dosimetry of the setup with four newborn mice properly placed in proximity of the antenna patch was carried out, exploiting a matching technique to decrease the reflections due to dielectric discontinuities (i.e., from air to mouse tissues). Such technique consists in the use of a matching dielectric box with dielectric permittivity similar to those of the mice. The average computed electric field inside single mice was homogeneous (better than 68 %) with an efficiency higher than 20 V m^?1 V^?1 for the four exposed mice. These results demonstrate the possibility of a multiple (four) exposure of small animals to short nanosecond pulses (both monopolar and bipolar) in a controlled and efficient way.     

Micronuclei formation in somatic cells of mice exposed to 50-Hz electric fields

Male Swiss mice are tested under uniform 50-Hz electric field intensities of 100, 170, 220, and 290 kV/m. These values on the basis of equal induced current density are equivalent to the case of a human exposed to field intensities of nearly 8, 14, 18, and 24 kV/m, respectively. The latter values may be found beneath or in the vicinity of extra-high-voltage power lines whose voltages range from 220 to 765 kV. The cytogenetic effect of extremely low-frequency (ELF) electric fields, as expressed by micronuclei formation, is assessed. Mice are exposed for 24 hr, and samples are taken 48, 72, and 96 hr from the beginning of exposure. Sham-exposed mice served as controls. The number of micronucleated polychromatic erythrocytes (PCE) in exposed animals are significantly higher than those of the control. The increase in micronucleated PCE was significantly dose dependent at all times. Samples taken 96 hr after exposure show a decrease in percentages of micronucleated PCE, which may be taken as an indication of recovery.     

Network and intrinsic contributions to carbachol-induced oscillations in the rat subiculum

Low-frequency network oscillations occur in several areas of the limbic system where they contribute to synaptic plasticity and mnemonic functions that are in turn modulated by cholinergic mechanisms. Here we used slices of the rat subiculum (a limbic area involved in cognitive functions) to establish how network and single neuron (intrinsic) membrane mechanisms participate to the rhythmic oscillations elicited by the cholinergic agent carbachol (CCh, 50-100 microM). We have found that CCh-induced network oscillations (intraoscillatory frequency = 5-16 Hz) are abolished by an antagonist of non-N-methyl-D-aspartate (NMDA) glutamatergic receptors (n = 6 slices) but persist during blockade of GABA receptors (n = 16). In addition, during application of glutamate and GABA receptor antagonists, single subicular cells generate burst oscillations at 2.1-6.8 Hz when depolarized with steady current injection. These intrinsic burst oscillations disappear during application of a Ca(2+) channel blocker (n = 6 cells), intracellular Ca(2+) chelation (n = 6), or replacement of extracellular Na(+) (n = 4) but persist in recordings made with electrodes containing a blocker of voltage-gated Na(+) channels (n = 7). These procedures cause similar effects on CCh-induced depolarizing plateau potentials that are contributed by a Ca(2+)-activated nonselective cationic conductance (I(CAN)). Network and intrinsic oscillations along with depolarizing plateau potentials were abolished by the muscarinic receptor antagonist atropine. In conclusion, our findings demonstrate that low-frequency oscillations in the rat subiculum rely on the muscarinic receptor-dependent activation of an intrinsic oscillatory mechanism that is presumably contributed by I(CAN) and are integrated within the network via non-NMDA receptor-mediated transmission.     

Hippocampal sharp wave-ripples linked to slow oscillations in rat slow-wave sleep

Slow oscillations originating in the prefrontal neocortex during slow-wave sleep (SWS) group neuronal network activity and thereby presumably support the consolidation of memories. Here, we investigated whether the grouping influence of slow oscillations extends to hippocampal sharp wave-ripple (SPW) activity thought to underlie memory replay processes during SWS. The prefrontal surface EEG and multiunit activity (MUA), along with hippocampal local field potentials (LFP) from CA1, were recorded in rats during sleep. Average spindle and ripple activity and event correlation histograms of SPWs were calculated, time-locked to half-waves of slow oscillations. Results confirm decreased prefrontal MUA and spindle activity during EEG slow oscillation negativity and increases in this activity during subsequent positivity. A remarkably close temporal link was revealed between slow oscillations and hippocampal activity, with ripple activity and SPWs being also distinctly decreased during negative half-waves and increased during slow oscillation positivity. Fine-grained analyses of temporal dynamics revealed for the slow oscillation a phase delay of approximately 90 ms with reference to up and down states of prefrontal MUA, and of only approximately 60 ms with reference to changes in SPWs, indicating that up and down states in prefrontal MUA precede corresponding changes in hippocampal SPWs by approximately 30 ms. Results support the notion that the depolarizing surface-positive phase of the slow oscillation and the associated up state of prefrontal excitation promotes hippocampal SPWs via efferent pathways. The preceding disfacilitation of hippocampal events temporally coupled to the negative slow oscillation half-wave appears to serve a synchronizing role in this neocorticohippocampal interplay.