Matthew I. Banks

Interactions between Distinct GABAA Circuits in Hippocampus

Synchronous activity among synaptically connected interneurons is thought to organize temporal patterns such as gamma and theta rhythms in cortical circuits. Interactions between distinct interneuron circuits may underlie more complex patterns, such as nested rhythms. Here, we demonstrate such an interaction between two groups of CA1 interneurons, GABA(A,slow) and GABA(A,fast) cells, that may contribute to theta and gamma rhythms, respectively. Stratum lacunosum-moleculare (SL-M) stimuli that activate GABA(A,slow) inhibitory postsynaptic currents (IPSCs) in pyramidal cells simultaneously depress the rate and amplitude of spontaneous GABA(A,fast) IPSCs for several hundred milliseconds. This suppression has a similar pharmacological profile to GABA(A,slow) IPSCs, and SL-M stimuli elicit GABA(A,slow) IPSCs in interneurons. We conclude that GABA(A,slow) cells inhibit both pyramidal cells and GABA(A,fast) interneurons and postulate that this interaction contributes to nested theta/gamma rhythms in hippocampus.

Responses of Cochlear Nucleus Cells and Projections of their Axons

The advent of intracellular recording using horseradish peroxidase-filled glass electrodes offered a new and exciting approach to the in vivo cochlear nucleus (CN) preparation. Sharp glass microelectrodes, filled with a standard salt solution containing positively charged HRP molecules, made it feasible to record and characterize supra- and subthreshold intracellular responses of individual neurons to auditory stimuli, inject and subsequently recover the same HRP-labeled cell and ask the following basic questions; 1) Do morphologically defined cell types in the cochlear nucleus respond in a certain way to simple auditory stimuli at both the sub- and suprathreshold level? 2) Are the synaptic inputs of different cell types different, in terms of location, type and concentration, and are they arranged in ways that might help to explain the cell type’s unique responsiveness? 3) What is the projection pattern of individual axons originating from a given cell type, what are the shapes of vesicles within the terminals of these axons and can we make educated guesses about their influence on other cell populations based on this information?

Thalamocortical projections to rat auditory cortex from the ventral and dorsal divisions of the medial geniculate nucleus

The ventral and dorsal medial geniculate (MGV and MGD) constitute the major auditory thalamic subdivisions providing thalamocortical inputs to layer IV and lower layer III of auditory cortex. No quantitative evaluation of this projection is available. Using biotinylated dextran amine (BDA)/biocytin injections, we describe the cortical projection patterns of MGV and MGD cells. In primary auditory cortex the bulk of MGV axon terminals are in layer IV/lower layer III with minor projections to supragranular layers and intermediate levels in infragranular layers. MGD axons project to cortical regions designated posterodorsal (PD) and ventral (VA) showing laminar terminal distributions that are quantitatively similar to the MGV‐to‐primary cortex terminal distribution. At the electron microscopic level MGV and MGD terminals are non‐γ‐aminobutyric acid (GABA)ergic with MGD terminals in PD and VA slightly but significantly larger than MGV terminals in primary cortex. MGV/MGD terminals synapse primarily onto non‐GABAergic spines/dendrites. A small number synapse on GABAergic structures, contacting large dendrites or cell bodies primarily in the major thalamocortical recipient layers. For MGV projections to primary cortex or MGD projections to PD or VA, the non‐GABAergic postsynaptic structures at each site were the same size regardless of whether they were in supragranular, granular, or infragranular layers. However, the population of MGD terminal‐recipient structures in VA were significantly larger than the MGD terminal‐recipient structures in PD or the MGV terminal‐recipient structures in primary cortex. Thus, if terminal and postsynaptic structure size indicate strength of excitation then MGD to VA inputs are strongest, MGD to PD intermediate, and MGV to primary cortex the weakest. J. Comp. Neurol., 2012.

Muscarinic blockade weakens interaction of gamma with theta rhythms in mouse hippocampus.

θ (4–12 Hz) and γ (40–90) oscillations are prominent rhythms in the mammalian brain. A striking feature of these rhythms, possibly vital to memory encoding, is their specific coordination in a manner that has been termed ‘nesting’, i.e. the preferred occurrence of bouts of γ activity during specific phases of θ. Both rhythms are shaped by the neuromodulator acetylcholine, but it is unknown to what degree their coordination is influenced by cholinergic neuromodulation. Here, we investigated the effects of a blockade of muscarinic acetylcholine receptors by atropine on θ and γ oscillations, and their interaction, in mouse hippocampus in vivo. Multi‐site recordings from area CA1 of freely moving mice showed that under control conditions γ activity was amplitude‐modulated at θ frequencies. This coordination of θ and γ oscillations, as assessed by cross‐correlation of θ with the γ envelope, was prominent in basal and apical dendritic laminae but not in intermediate laminae. It was stronger during active exploration than during awake immobility. Atropine (50 mg/kg intraperitoneal) altered several aspects of the individual and nested rhythms. It rendered θ activity irregular, decreased θ oscillation frequency and reduced γ power. Atropine also reduced the amplitude‐modulation of γ oscillations at θ frequencies, in part by perturbing the coordination of the rhythms on a short time scale. Thus, our findings demonstrate that phase locking of the amplitude of γ oscillations to θ in hippocampal area CA1 is partially governed by neuronal elements harbouring muscarinic receptors.

Bistable Network Behavior of Layer I Interneurons in Auditory Cortex

GABAergic interneurons in many areas of the neocortex are mutually connected via chemical and electrical synapses. Previous computational studies have explored how these coupling parameters influence the firing patterns of interneuronal networks. These models have predicted that the stable states of such interneuronal networks will be either synchrony (near zero phase lag) or antisynchrony (phase lag near one-half of the interspike interval), depending on network connectivity and firing rates. In certain parameter regimens, the network can be bistable, settling into either stable state depending on the initial conditions. Here, we investigated how connectivity parameters influence spike patterns in paired recordings from layer I interneurons in brain slices from juvenile mice. Observed properties of chemical and electrical synapses were used to simulate connections between uncoupled cells via dynamic clamp. In uncoupled pairs, action potentials induced by constant depolarizing currents had randomly distributed phase differences between the two cells. When coupled with simulated chemical (inhibitory) synapses, however, these pairs exhibited a bimodal firing pattern, tending to fire either in synchrony or in antisynchrony. Combining electrical with chemical synapses, prolonging τDecay of inhibitory connections, or increasing the firing rate of the network all resulted in enhanced stability of the synchronous state. Thus, electrical and inhibitory synaptic coupling constrain the relative timing of spikes in a two-cell network to, at most, two stable states, the stability and precision of which depend on the exact parameters of coupling.

Influence of GABAA receptor γ2 splice variants on receptor kinetics and isoflurane modulation

Background:γ-Aminobutyric acid type A (GABAA) receptors, the major inhibitory receptors in the brain, are important targets of many drugs, including general anesthetics. These compounds exert multiple effects on GABAA receptors, including direct activation, prolongation of deactivation kinetics, and reduction of inhibitory postsynaptic current amplitudes. However, the degree to which these actions occur differs for different agents and synapses, possibly because of subunit-specific effects on postsynaptic receptors. In contrast to benzodiazepines and intravenous anesthetics, there is little information available about the subunit dependency of actions of volatile anesthetics. Therefore, the authors studied in detail the effects of isoflurane on recombinant GABAA receptors composed of several different subunit combinations. Methods:Human embryonic kidney 293 cells were transiently transfected with rat complementary DNAs of α1β2, α1β2γ2L, α1β2γ2S, α5β3, or α5β3γ2S subunits. Using rapid application and whole cell patch clamp techniques, cells were exposed to 10- and 2,000-ms pulses of γ-aminobutyric acid (1 mm) in the presence or absence of isoflurane (0.25, 0.5, 1.0 mm). Anesthetic effects on decay kinetics, peak amplitude, net charge transfer and rise time were measured. Statistical significance was assessed using the Student t test or one-way analysis of variance followed by the Tukey post hoc test. Results:Under control conditions, incorporation of a γ2 subunit conferred faster deactivation kinetics and reduced desensitization. Isoflurane slowed deactivation, enhanced desensitization, and reduced peak current amplitude in αβ receptors. Coexpression with a γ2 subunit caused these effects of isoflurane to be substantially reduced or abolished. Although the two γ2 splice variants imparted qualitatively similar macroscopic kinetic properties, there were significant quantitative differences between effects of isoflurane on deactivation and peak current amplitude in γ2S- versus γ2L-containing receptors. The net charge transfer resulting from brief pulses of γ-aminobutyric acid was decreased by isoflurane in αβ but increased in αβγ receptors. Conclusions:The results indicate that subunit composition does substantially influence modulation of GABAA receptors by isoflurane. Specifically, the presence of a γ2 subunit and the identity of its splice variant are important factors in determining physiologic and pharmacologic properties. These results may have functional implications in understanding how anesthetic effects on specific types of GABAA receptors in the brain contribute to changes in brain function and behavior.

Preferential effect of isoflurane on top-down vs. bottom-up pathways in sensory cortex

The mechanism of loss of consciousness (LOC) under anesthesia is unknown. Because consciousness depends on activity in the cortico-thalamic network, anesthetic actions on this network are likely critical for LOC. Competing theories stress the importance of anesthetic actions on bottom-up “core” thalamo-cortical (TC) vs. top-down cortico-cortical (CC) and matrix TC connections. We tested these models using laminar recordings in rat auditory cortex in vivo and murine brain slices. We selectively activated bottom-up vs. top-down afferent pathways using sensory stimuli in vivo and electrical stimulation in brain slices, and compared effects of isoflurane on responses evoked via the two pathways. Auditory stimuli in vivo and core TC afferent stimulation in brain slices evoked short latency current sinks in middle layers, consistent with activation of core TC afferents. By contrast, visual stimuli in vivo and stimulation of CC and matrix TC afferents in brain slices evoked responses mainly in superficial and deep layers, consistent with projection patterns of top-down afferents that carry visual information to auditory cortex. Responses to auditory stimuli in vivo and core TC afferents in brain slices were significantly less affected by isoflurane compared to responses triggered by visual stimuli in vivo and CC/matrix TC afferents in slices. At a just-hypnotic dose in vivo, auditory responses were enhanced by isoflurane, whereas visual responses were dramatically reduced. At a comparable concentration in slices, isoflurane suppressed both core TC and CC/matrix TC responses, but the effect on the latter responses was far greater than on core TC responses, indicating that at least part of the differential effects observed in vivo were due to local actions of isoflurane in auditory cortex. These data support a model in which disruption of top-down connectivity contributes to anesthesia-induced LOC, and have implications for understanding the neural basis of consciousness.

Descending Projections from Extrastriate Visual Cortex Modulate Responses of Cells in Primary Auditory Cortex

Primary sensory cortical responses are modulated by the presence or expectation of related sensory information in other modalities, but the sources of multimodal information and the cellular locus of this integration are unclear. We investigated the modulation of neural responses in the murine primary auditory cortical area Au1 by extrastriate visual cortex (V2). Projections from V2 to Au1 terminated in a classical descending/modulatory pattern, with highest density in layers 1, 2, 5, and 6. In brain slices, whole-cell recordings revealed long latency responses to stimulation in V2L that could modulate responses to subsequent white matter (WM) stimuli at latencies of 5-20 ms. Calcium responses imaged in Au1 cell populations showed that preceding WM with V2L stimulation modulated WM responses, with both summation and suppression observed. Modulation of WM responses was most evident for near-threshold WM stimuli. These data indicate that corticocortical projections from V2 contribute to multimodal integration in primary auditory cortex.

Modulation of γ-Aminobutyric Acid Type A Receptor-mediated Spontaneous Inhibitory Postsynaptic Currents in Auditory Cortex by Midazolam and Isoflurane

Background: Anesthetic agents that target &ggr;-aminobutyric acid type A (GABAA) receptors modulate cortical auditory evoked responses in vivo, but the cellular targets involved are unidentified. Also, for agents with multiple protein targets, the relative contribution of modulation of GABAA receptors to effects on cortical physiology is unclear. The authors compared effects of the GABAA receptor–specific drug midazolam with the volatile anesthetic isoflurane on spontaneous inhibitory postsynaptic currents (sIPSCs) in pyramidal cells of auditory cortex. Methods: Whole cell recordings were obtained in murine brain slices at 34°C. GABAA sIPSCs were isolated by blocking ionotropic glutamate receptors. Effects of midazolam and isoflurane on time course, amplitude, and frequency of sIPSCs were measured. Results: The authors detected no effect of midazolam at 0.01 &mgr;m on sIPSCs, whereas midazolam at 0.1 and 1 &mgr;m prolonged the decay of sIPSCs by approximately 25 and 70%, respectively. Isoflurane at 0.1, 0.25, and 0.5 mm prolonged sIPSCs by approximately 45, 150, and 240%, respectively. No drug-specific effects were observed on rise time or frequency of sIPSCs. Isoflurane at 0.5 mm caused a significant decrease in sIPSC amplitude. Conclusions: The dose dependence of isoflurane effects on GABAA sIPSCs in pyramidal cells is consistent with effects on auditory evoked response in vivo. By contrast, comparable effects of midazolam on GABAA sIPSCs arise at concentrations exceeding those currently thought to be achieved in vivo, suggesting that the cellular targets of midazolam reside elsewhere in the thalamocortical circuit or that the concentration of midazolam reached in the brain is higher than currently believed.

Spiking in auditory cortex following thalamic stimulation is dominated by cortical network activity.

The state of the sensory cortical network can have a profound impact on neural responses and perception. In rodent auditory cortex, sensory responses are reported to occur in the context of network events, similar to brief UP states, that produce “packets” of spikes and are associated with synchronized synaptic input (Bathellier et al., 2012; Hromadka et al., 2013; Luczak et al., 2013). However, traditional models based on data from visual and somatosensory cortex predict that ascending sensory thalamocortical (TC) pathways sequentially activate cells in layers 4 (L4), L2/3, and L5. The relationship between these two spatio-temporal activity patterns is unclear. Here, we used calcium imaging and electrophysiological recordings in murine auditory TC brain slices to investigate the laminar response pattern to stimulation of TC afferents. We show that although monosynaptically driven spiking in response to TC afferents occurs, the vast majority of spikes fired following TC stimulation occurs during brief UP states and outside the context of the L4>L2/3>L5 activation sequence. Specifically, monosynaptic subthreshold TC responses with similar latencies were observed throughout layers 2–6, presumably via synapses onto dendritic processes located in L3 and L4. However, monosynaptic spiking was rare, and occurred primarily in L4 and L5 non-pyramidal cells. By contrast, during brief, TC-induced UP states, spiking was dense and occurred primarily in pyramidal cells. These network events always involved infragranular layers, whereas involvement of supragranular layers was variable. During UP states, spike latencies were comparable between infragranular and supragranular cells. These data are consistent with a model in which activation of auditory cortex, especially supragranular layers, depends on internally generated network events that represent a non-linear amplification process, are initiated by infragranular cells and tightly regulated by feed-forward inhibitory cells.