神経細胞のマルチコンパートメントモデルにおける細胞外抵抗分布の役割 : 細胞外記録および細胞外電気刺激のための統一形式化

Tohoku University川島隆

Local Field Potential Modeling Predicts Dense Activation in Cerebellar Granule Cells Clusters under LTP and LTD Control

Local field-potentials (LFPs) are generated by neuronal ensembles and contain information about the activity of single neurons. Here, the LFPs of the cerebellar granular layer and their changes during long-term synaptic plasticity (LTP and LTD) were recorded in response to punctate facial stimulation in the rat in vivo. The LFP comprised a trigeminal (T) and a cortical (C) wave. T and C, which derived from independent granule cell clusters, co-varied during LTP and LTD. To extract information about the underlying cellular activities, the LFP was reconstructed using a repetitive convolution (ReConv) of the extracellular potential generated by a detailed multicompartmental model of the granule cell. The mossy fiber input patterns were determined using a Blind Source Separation (BSS) algorithm. The major component of the LFP was generated by the granule cell spike Na+ current, which caused a powerful sink in the axon initial segment with the source located in the soma and dendrites. Reproducing the LFP changes observed during LTP and LTD required modifications in both release probability and intrinsic excitability at the mossy fiber-granule cells relay. Synaptic plasticity and Golgi cell feed-forward inhibition proved critical for controlling the percentage of active granule cells, which was 11% in standard conditions but ranged from 3% during LTD to 21% during LTP and raised over 50% when inhibition was reduced. The emerging picture is that of independent (but neighboring) trigeminal and cortical channels, in which synaptic plasticity and feed-forward inhibition effectively regulate the number of discharging granule cells and emitted spikes generating “dense” activity clusters in the cerebellar granular layer

A biophysical observation model for field potentials of networks of leaky integrate-and-fire neurons

We present a biophysical approach for the coupling of neural network activity as resulting from proper dipole currents of cortical pyramidal neurons to the electric field in extracellular fluid. Starting from a reduced threecompartment model of a single pyramidal neuron, we derive an observation model for dendritic dipole currents in extracellular space and thereby for the dendritic field potential that contributes to the local field potential of a neural population. This work aligns and satisfies the widespread dipole assumption that is motivated by the "open-field" configuration of the dendritic field potential around cortical pyramidal cells. Our reduced three-compartment scheme allows to derive networks of leaky integrate-and-fire models, which facilitates comparison with existing neural network and observation models. In particular, by means of numerical simulations we compare our approach with an ad hoc model by Mazzoni et al. [Mazzoni, A., S. Panzeri, N. K. Logothetis, and N. Brunel (2008). Encoding of naturalistic stimuli by local field potential spectra in networks of excitatory and inhibitory neurons. PLoS Computational Biology 4 (12), e1000239], and conclude that our biophysically motivated approach yields substantial improvement.Comment: 31 pages, 4 figure

Disentangling astroglial physiology with a realistic cell model in silico

Electrically non-excitable astroglia take up neurotransmitters, buffer extracellular K+ and generate Ca2+ signals that release molecular regulators of neural circuitry. The underlying machinery remains enigmatic, mainly because the sponge-like astrocyte morphology has been difficult to access experimentally or explore theoretically. Here, we systematically incorporate multi-scale, tri-dimensional astroglial architecture into a realistic multi-compartmental cell model, which we constrain by empirical tests and integrate into the NEURON computational biophysical environment. This approach is implemented as a flexible astrocyte-model builder ASTRO. As a proof-of-concept, we explore an in silico astrocyte to evaluate basic cell physiology features inaccessible experimentally. Our simulations suggest that currents generated by glutamate transporters or K+ channels have negligible distant effects on membrane voltage and that individual astrocytes can successfully handle extracellular K+ hotspots. We show how intracellular Ca2+ buffers affect Ca2+ waves and why the classical Ca2+ sparks-and-puffs mechanism is theoretically compatible with common readouts of astroglial Ca2+ imaging

Spike-Feature Based Estimation of Electrode Position in Extracellular Neural Recordings

Detecting and sorting spikes in extracellular neural recordings are common procedures in assessing the activity of individual neurons. In chronic recordings, passive electrode movements introduce changes in the shape of detected spike waveforms, and may thus lead to problems with identification and tracking of spikes recorded at separate instances in time, which is an important step in long-term monitoring of individual neurons. Information about electrode movements after implantation is crucial to the evaluation of mechanical stability of different electrode designs. In this paper, we present a preliminary study of the relationship between electrode movements and the resulting movements of spike-features in feature space. We show that there is a characteristic relationship between the two movements and that this relationship can be modeled as a linear transformation between two coordinate systems. Finally, we show how the relationship can be used for estimating electrode positions based on measured spike waveforms without any prior knowledge about the type of neuron by introducing a learning procedure during electrode insertion

Frequency dependence of signal power and spatial reach of the local field potential

The first recording of electrical potential from brain activity was reported already in 1875, but still the interpretation of the signal is debated. To take full advantage of the new generation of microelectrodes with hundreds or even thousands of electrode contacts, an accurate quantitative link between what is measured and the underlying neural circuit activity is needed. Here we address the question of how the observed frequency dependence of recorded local field potentials (LFPs) should be interpreted. By use of a well-established biophysical modeling scheme, combined with detailed reconstructed neuronal morphologies, we find that correlations in the synaptic inputs onto a population of pyramidal cells may significantly boost the low-frequency components of the generated LFP. We further find that these low-frequency components may be less `local' than the high-frequency LFP components in the sense that (1) the size of signal-generation region of the LFP recorded at an electrode is larger and (2) that the LFP generated by a synaptically activated population spreads further outside the population edge due to volume conduction

High-resolution three-dimensional extracellular recording of neuronal activity with microfabricated electrode arrays

Microelectrode array recordings of neuronal activity present significant opportunities for studying the brain with single-cell and spike-time precision. However, challenges in device manufacturing constrain dense multisite recordings to two spatial dimensions, whereas access to the three-dimensional (3D) structure of many brain regions appears to remain a challenge. To overcome this limitation, we present two novel recording modalities of silicon-based devices aimed at establishing 3D functionality. First, we fabricated a dual-side electrode array by patterning recording sites on both the front and back of an implantable microstructure. We found that the majority of single-unit spikes could not be simultaneously detected from both sides, suggesting that in addition to providing higher spatial resolution measurements than that of single-side devices, dual-side arrays also lead to increased recording yield. Second, we obtained recordings along three principal directions with a multilayer array and demonstrated 3D spike source localization within the enclosed measurement space. The large-scale integration of such dual-side and multilayer arrays is expected to provide massively parallel recording capabilities in the brain

Investigation Of The Spatiotemporal Dynamics Of Camp And Pka Signaling And The Role Of Hcn4 Subunits In Anxiety-Related Behavior And Memory

In the hippocampus, long-term memory and synaptic plasticity occur through a series of coordinated intracellular signaling cascades that strengthen and stabilize subsets of synaptic connections while leaving thousands of others unaltered. Therefore, understanding how molecular signals are accurately transmitted is critical to understanding how hippocampal neurons store information. Molecules like cAMP and protein kinase A are critical components of memory and plasticity, but it is unclear how these diffusible signals are dynamically regulated to achieve the spatial and temporal specificity that underlies pathway-specific plasticity. Hyperpolarization-activated and cyclic nucleotide-gated (HCN) channels are ion channels that are modulated by cAMP and are known to regulate the spatial and temporal dynamics of excitatory postsynaptic potentials. HCN1 and HCN2 subunits have been implicated in memory, plasticity and anxiety-related behaviors, but the role for HCN4 subunits remains untested. In Chapter 1, I review the role of cAMP signaling in hippocampal synaptic plasticity and memory consolidation with emphasis on the molecular mechanisms regulating cAMP, PKA and HCN channels. In Chapter 2, I combine live two-photon imaging of genetically-encoded fluorescent FRET sensors and computational modeling to investigate the molecular mechanisms regulating the spatiotemporal dynamics of cAMP and PKA activity in hippocampal neurons during stimulation of β-adrenergic receptors. Results suggest that the ratio between adenylyl cyclase and phosphodiesterase-4 scales with neuronal compartment size to maintain basal cAMP levels and produce rapid-onset, high-amplitude cAMP transients in small compartments. Conversely, imaging experiments show that PKA activity is greater in large neuronal compartments and modeling suggests that compartmental differences in PKA activity depend on the concentration of protein phosphatase and not on the concentration of PKA substrates or PKA holoenzyme. In Chapter 3, I use recombinant adeno-associated viruses and shRNA-mediated silencing of HCN4 subunits to examine their role in anxiety, memory, and contextual fear extinction. Results from a battery of behavioral assays suggest that reduction of HCN4 subunits increases anxiety-related behavior, but does not affect object-location memory or contextual fear conditioning. Together, my thesis work provides novel insight into the molecular mechanism regulating the spatiotemporal dynamics of cAMP/PKA signaling and provides suggests a role for HCN4 subunits in anxiety-related behavior

Dual-Compartment Neurofluidic System for Electrophysiological Measurements in Physically Segregated and Functionally Connected Neuronal Cell Culture

We developed a dual-compartment neurofluidic system with inter-connecting microchannels to connect neurons from their respective compartments, placed on a planar microelectrode arrays. The design and development of the compartmented microfluidic device for neuronal cell culture, protocol for sustaining long-term cultures, and neurite growth through microchannels in such a closed compartment device are presented. Using electrophysiological measurements of spontaneous network activity in the compartments and selective pharmacological manipulation of cells in one compartment, the biological origin of network activity and the fluidic isolation between the compartments are demonstrated. The connectivity between neuronal populations via the microchannels and the crossing-over of neurites are verified using transfection experiments and immunofluorescence staining. In addition to the neurite cross-over to the adjacent compartment, functional connectivity between cells in both the compartments is verified using cross-correlation (CC) based techniques. Bidirectional signal propagation between the compartments is demonstrated using functional connectivity maps. CC analysis and connectivity maps demonstrate that the two neuronal populations are not only functionally connected within each compartment but also with each other and a well connected functional network was formed between the compartments despite the physical barrier introduced by the microchannels