Andrea Bibbig

A Possible Role for Gap Junctions in Generation of Very Fast EEG Oscillations Preceding the Onset of, and Perhaps Initiating, Seizures

Summary:  Purpose: We propose an experimentally and clinically testable hypothesis, concerning the origin of very fast (>∼70 Hz) EEG oscillations that sometimes precede the onset of focal seizures. These oscillations are important, as they may play a causal role in the initiation of seizures.

A model of gamma-frequency network oscillations induced in the rat CA3 region by carbachol in vitro

Carbachol (> 20 μm) and kainate (100 nm) induce, in the in vitro CA3 region, synchronized neuronal population oscillations at ≈ 40 Hz having distinctive features: (i) the oscillations persist for hours; (ii) interneurons in kainate fire at 5–20 Hz and their firing is tightly locked to field potential maxima (recorded in s. radiatum); (iii) in contrast, pyramidal cells, in both carbachol and kainate, fire at frequencies as low as 2 Hz, and their firing is less tightly locked to field potentials; (iv) the oscillations require GABAA receptors, AMPA receptors and gap junctions. Using a network of 3072 pyramidal cells and 384 interneurons (each multicompartmental and containing a segment of unmyelinated axon), we employed computer simulations to examine conditions under which network oscillations might occur with the experimentally determined properties. We found that such network oscillations could be generated, robustly, when gap junctions were located between pyramidal cell axons, as suggested to occur based on studies of spontaneous high‐frequency (> 100 Hz) network oscillations in the in vitro hippocampus. In the model, pyramidal cell somatic firing was not essential for the oscillations. Critical components of the model are (i) the plexus of pyramidal cell axons, randomly and sparsely interconnected by gap junctions; (ii) glutamate synapses onto interneurons; (iii) synaptic inhibition between interneurons and onto pyramidal cell axons and somata; (iv) a sufficiently high rate of spontaneous action potentials generated in pyramidal cell axons. This model explains the dependence of network oscillations on GABAA and AMPA receptors, as well as on gap junctions. Besides the existence of axon–axon gap junctions, the model predicts that many of the pyramidal cell action potentials, during sustained gamma oscillations, are initiated in axons.

A beta2-frequency (20-30 Hz) oscillation in nonsynaptic networks of somatosensory cortex.

Beta2 frequency (20–30 Hz) oscillations appear over somatosensory and motor cortices in vivo during motor preparation and can be coherent with muscle electrical activity. We describe a beta2 frequency oscillation occurring in vitro in networks of layer V pyramidal cells, the cells of origin of the corticospinal tract. This beta2 oscillation depends on gap junctional coupling, but it survives a cut through layer 4 and, hence, does not depend on apical dendritic electrogenesis. It also survives a blockade of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors or a blockade of GABAA receptors that is sufficient to suppress gamma (30–70 Hz) oscillations in superficial cortical layers. The oscillation period is determined by the M type of K+ current.

Axonal Gap Junctions Between Principal Neurons: A Novel Source of Network Oscillations, and Perhaps Epileptogenesis

We hypothesized in 1998 that gap junctions might be located between the axons of principal hippocampal neurons, based on the shape of spikelets (fast prepotentials), occurring during gap junction-mediated very fast (to approximately 200 Hz) network oscillations in vitro. More recent electrophysiological, pharmacological and dye-coupling data indicate that axonal gap junctions exist; so far, they appear to be located about 100 microm from the soma, in CA1 pyramidal neurons. Computer modeling and theory predict that axonal gap junctions can lead to very fast network oscillations under three conditions: a) there are spontaneous axonal action potentials; b) the number of gap junctions in the network is neither too low (not less than to approximately 1.5 per cell on average), nor too high (not more than to approximately 3 per cell on average); c) action potentials can cross from axon to axon via gap junctions. Simulated oscillations resemble biological ones, but condition (c) remains to be demonstrated directly. Axonal network oscillations can, in turn, induce oscillatory activity in larger neuronal networks, by a variety of mechanisms. Axonal networks appear to underlie in vivo ripples (to approximately 200 Hz field potential oscillations superimposed on physiological sharp waves), to drive gamma (30-70 Hz) oscillations that appear in the presence of carbachol, and to initiate certain types of ictal discharge. If axonal gap junctions are important for seizure initiation in humans, there could be practical consequences for antiepileptic therapy: at least one gap junction-blocking compound, carbenoxolone, is already in clinical use (for treatment of ulcer disease), and it crosses the blood-brain barrier.

GABA-enhanced collective behavior in neuronal axons underlies persistent gamma-frequency oscillations

Gamma (30–80 Hz) oscillations occur in mammalian electroencephalogram in a manner that indicates cognitive relevance. In vitro models of gamma oscillations demonstrate two forms of oscillation: one occurring transiently and driven by discrete afferent input and the second occurring persistently in response to activation of excitatory metabotropic receptors. The mechanism underlying persistent gamma oscillations has been suggested to involve gap-junctional communication between axons of principal neurons, but the precise relationship between this neuronal activity and the gamma oscillation has remained elusive. Here we demonstrate that gamma oscillations coexist with high-frequency oscillations (>90 Hz). High-frequency oscillations can be generated in the axonal plexus even when it is physically isolated from pyramidal cell bodies. They were enhanced in networks by nonsomatic γ-aminobutyric acid type A (GABAA) receptor activation, were modulated by perisomatic GABAA receptor-mediated synaptic input to principal cells, and provided the phasic input to interneurons required to generate persistent gamma-frequency oscillations. The data suggest that high-frequency oscillations occurred as a consequence of random activity within the axonal plexus. Interneurons provide a mechanism by which this random activity is both amplified and organized into a coherent network rhythm.

Contrasting roles of axonal (pyramidal cell) and dendritic (interneuron) electrical coupling in the generation of neuronal network oscillations

Electrical coupling between pyramidal cell axons, and between interneuron dendrites, have both been described in the hippocampus. What are the functional roles of the two types of coupling? Interneuron gap junctions enhance synchrony of γ oscillations (25–70 Hz) in isolated interneuron networks and also in networks containing both interneurons and principal cells, as shown in mice with a knockout of the neuronal (primarily interneuronal) connexin36. We have recently shown that pharmacological gap junction blockade abolishes kainate-induced γ oscillations in connexin36 knockout mice; without such gap junction blockade, γ oscillations do occur in the knockout mice, albeit at reduced power compared with wild-type mice. As interneuronal dendritic electrical coupling is almost absent in the knockout mice, these pharmacological data indicate a role of axonal electrical coupling in generating the γ oscillations. We construct a network model of an experimental γ oscillation, known to be regulated by both types of electrical coupling. In our model, axonal electrical coupling is required for the γ oscillation to occur at all; interneuron dendritic gap junctions exert a modulatory effect.

Genetically altered AMPA-type glutamate receptor kinetics in interneurons disrupt long-range synchrony of gamma oscillation.

Gamma oscillations synchronized between distant neuronal populations may be critical for binding together brain regions devoted to common processing tasks. Network modeling predicts that such synchrony depends in part on the fast time course of excitatory postsynaptic potentials (EPSPs) in interneurons, and that even moderate slowing of this time course will disrupt synchrony. We generated mice with slowed interneuron EPSPs by gene targeting, in which the gene encoding the 67-kDa form of glutamic acid decarboxylase (GAD67) was altered to drive expression of the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) glutamate receptor subunit GluR-B. GluR-B is a determinant of the relatively slow EPSPs in excitatory neurons and is normally expressed at low levels in γ-aminobutyric acid (GABA)ergic interneurons, but at high levels in the GAD-GluR-B mice. In both wild-type and GAD-GluR-B mice, tetanic stimuli evoked gamma oscillations that were indistinguishable in local field potential recordings. Remarkably, however, oscillation synchrony between spatially separated sites was severely disrupted in the mutant, in association with changes in interneuron firing patterns. The congruence between mouse and model suggests that the rapid time course of AMPA receptor-mediated EPSPs in interneurons might serve to allow gamma oscillations to synchronize over distance.

Persistent gamma oscillations in superficial layers of rat auditory neocortex: experiment and model.

Persistent in vitro gamma oscillations, induced by bath application of carbachol and kainate (amongst other drugs), were discovered by Eberhard Buhl and collaborators in 1998. The oscillations are robust, in that they can continue for hours; but the oscillations are also intricate in their mechanisms: they depend upon phasic synaptic excitation and inhibition, upon electrical coupling between interneurones and between pyramidal neurones, and – at least in neocortex – they depend upon complex intrinsic properties of some of the neurones.

Gap Junctions, Fast Oscillations and the Initiation of Seizures

In this chapter, we shall review evidence that gap junctions can contribute to epileptogenesis in the hippocampus and cortex—but not just any gap junctions. Rather, we shall argue for a role for a newly described sort of gap junction, located between the proximal axons of principal neurons. Such axon-axon gap junctions promote epileptogenesis not so much by enhancing synchrony, as by providing pathways for the direct spread of action potentials between neurons. A by-product of such spread is the ability of axonally-coupled neurons to generate oscillations at very high frequencies (>~70 Hz). It is of note that seizure activity, both in vivo and in vitro, has been observed to begin with very high-frequency oscillations. If such oscillations can be shown to initiate the seizure discharge, and not just be an epiphenomenon, then targeting gap junction conductances may prove useful as an anticonvulsant strategy.

Oscillation-modifying drugs modify cognition in Alzheimer's mouse models

factor in the quality of actual radiation delivery. The capability of generating an entire volumetric MV-CBCT data set in a single-gantry rotation, allows 3D visualization of the tumor prior to the delivery of treatment and correlation with reference plan CT data. This permits corrections of shifts beyond an acceptable limit Methods: Prior to treatment, 2D and/or CBCT on ARTISTE (M/s Siemens) was acquired and setup errors with reference to X, Y, Z were corrected online in 20 patients of breast, head & neck (H&N) and prostate. A second CBCTwas acquired after the correction process and coordinates for daily set-up and images were obtained. Results: A total number of 211 CBCT or 2D images were performed in 20 patients. The sites included breast (n1⁄410), H&N (n1⁄46) and prostate (n1⁄44). Images were evaluated for 95, 58 and 58 fractions respectively. The shifts observed in X, Y and Z axes are summarized below: In addition, rotational errors were observed in 7% (15/211 images). These include breast (2%), H&N (1%) and prostate (4%), which were also corrected by IGRT Conclusions: Despite immobilization devices, shifts beyond the acceptable limits of 2mm were observed during online CBCT or 2D imaging with IGRT in breast (79.9%), H&N (49.2%) and prostate (96.6%). IGRT permits detection and online corrections of these shifts which would have been otherwise gone unnoticed leading to dosimetric errors during radiation therapy.