Wrestling model of the repertoire of activity propagation modes in quadruple neural networks.

The spontaneous activity of engineered quadruple cultured neural networks (of four-coupled sub-networks) exhibits a repertoire of different types of mutual synchronization events. Each event corresponds to a specific activity propagation mode (APM) defined by the order of activity propagation between the sub-networks. We statistically characterized the frequency of spontaneous appearance of the different types of APMs. The relative frequencies of the APMs were then examined for their power-law properties. We found that the frequencies of appearance of the leading (most frequent) APMs have close to constant algebraic ratio reminiscent of Zipf's scaling of words. We show that the observations are consistent with a simplified "wrestling" model. This model represents an extension of the "boxing arena" model which was previously proposed to describe the ratio between the two activity modes in two coupled sub-networks. The additional new element in the "wrestling" model presented here is that the firing within each network is modeled by a time interval generator with similar intra-network Lévy distribution. We modeled the different burst-initiation zones' interaction by competition between the stochastic generators with Gaussian inter-network variability. Estimation of the model parameters revealed similarity across different cultures while the inter-burst-interval of the cultures was similar across different APMs as numerical simulation of the model predicts.

Antifreeze protein suppresses spontaneous neural activity and protects neurons from hypothermia/re-warming injury.

Antifreeze proteins (AFP) are associated with protection from freezing. We measured the effect of type I antifreeze protein on spontaneous bursting of mixed neuronal/glial cultures using a multi-electrode array culture system. Antifreeze protein (10mg/ml) reversibly depressed bursting activity without inhibiting mitochondrial oxidative capacity. The effect of antifreeze protein on cold/re-warming injury was investigated in rat hippocampal slice cultures. Compared to bovine serum albumin at a similar concentration, antifreeze protein protected hippocampal neurons from 8h of profound hypothermia at (4 degrees C) followed by re-warming. The protection observed is believed to be associated with the inhibitory effect of antifreeze protein.

The emergence and properties of mutual synchronization in in vitro coupled cortical networks.

We have studied the emergence of mutual synchronization and activity propagation in coupled neural networks from rat cortical cells grown on a micro-electrode array for parallel activity recording of dozens of neurons. The activity of each sub-network by itself is marked by the formation of synchronized bursting events (SBE) - short time windows of rapid neuronal firing. The joint activity of two coupled networks is characterized by the formation of mutual synchronization, i.e. the formation of SBE whose activity starts at one sub-network and then propagates to the other. The sub-networks switch roles in initiating the mutual SBE. However, spontaneous propagation (initiation) asymmetry emerges - one of the sub-networks takes on the role of initiating substantially more mutual SBE than the other, despite the fact that the two are engineered to be similar in size and cell density. Analysis of the interneuron correlations in the SBE also reveals the emergence of activity (function) asymmetry - one sub-network develops a more organized structure of correlations. We also show activity propagation and mutual synchronization in four coupled networks. Using computer simulations, we propose that the function asymmetry reflects asymmetry between the internal connectivity of the two networks, whereas the propagation asymmetry reflects asymmetry in the connectivity between the sub-networks. These results agree with the experimental findings that the initiation and function asymmetry can be separately regulated, which implies that information transfer (activity propagation) and information processing (function) can be regulated separately in coupled neural networks.

Management of synchronized network activity by highly active neurons.

Increasing evidence supports the idea that spontaneous brain activity may have an important functional role. Cultured neuronal networks provide a suitable model system to search for the mechanisms by which neuronal spontaneous activity is maintained and regulated. This activity is marked by synchronized bursting events (SBEs)--short time windows (hundreds of milliseconds) of rapid neuronal firing separated by long quiescent periods (seconds). However, there exists a special subset of rapidly firing neurons whose activity also persists between SBEs. It has been proposed that these highly active (HA) neurons play an important role in the management (i.e. establishment, maintenance and regulation) of the synchronized network activity. Here, we studied the dynamical properties and the functional role of HA neurons in homogeneous and engineered networks, during early network development, upon recovery from chemical inhibition and in response to electrical stimulations. We found that their sequences of inter-spike intervals (ISI) exhibit long time correlations and a unimodal distribution. During the network's development and under intense inhibition, the observed activity follows a transition period during which mostly HA neurons are active. Studying networks with engineered geometry, we found that HA neurons are precursors (the first to fire) of the spontaneous SBEs and are more responsive to electrical stimulations.

Spatio-temporal motifs 'remembered' in neuronal networks following profound hypothermia.

Surgical procedures using hypothermic temperatures have been linked to complications such as seizures, impaired mental development and impaired memory. Although there is some evidence that the profound hypothermia (<12 ( composite function)C) used in these procedures may be contributing to these neurological impairments, skepticism remains because of lack of evidence from experimental studies isolating the effects of hypothermia on neuronal networks. In order to attain a better understanding of profound hypothermia effects on neurons during surgical procedures, we applied cold to a cultured in-vitro neuronal network. The typical pattern of activity of such cultures is in the form of synchronized bursts, in which most of the recorded neurons fire action potentials in a short time period. In most cases, the bursting activity shows one or more repeating precise spatio-temporal patterns (motifs) that are sustained over long periods of time. In this experimental study, neuronal networks grown on microelectrode arrays (MEA) are subjected to profound hypothermia for an hour and the collective dynamics of the network as a whole are assessed. We show, by using a similarity analysis that compares changes in the time delays between neuronal activation at different burst motifs, that neuronal networks survive total inhibition by profound hypothermia and retain their intrinsic synchronized burst motifs even with substantial generalized neuronal degeneration. By applying multiple sessions of cold, we also show a marked monotonic reduction in the rate of burst firing and in the number of spikes of each neuron after each session.

Identifying repeating motifs in the activation of synchronized bursts in cultured neuronal networks.

Cultured neuronal networks cultivated on micro-electrode arrays are a widely used tool for the investigation of network mechanisms, providing structural framework for long-term recordings of network electrical activity, as well as the network reaction to electrical or chemical stimulations. The typical activity pattern of the culture takes the form of synchronized bursting events (SBEs), in which a large fraction of the recorded neurons simultaneously fire trains of action potentials in short bursts of several hundreds of a millisecond. We developed a method that identifies clusters of bursts that share a similar activation motif throughout the culture based on the fact that the culture morphology remains relatively unchanged for an extended time interval and that neurons fire in a recognizable and precise manner during a burst initiation. Our method compares accuracies in time delays that occurred between the activation of spike-trains of different neurons. Three culture architectures were studied and analyzed: a large network of 2 million cells, a smaller network limited in size of 100,000 cells, and a large network divided into 4 clusters. In each of the morphologies we identified cultures that showed more than one activation motif. Clustered networks showed more motifs on average than uniform cultures. The algorithm was able to show high fidelity to artificial noise. We also compare the results of our method with another method based on a correlation measure.

Study of hypothermia on cultured neuronal networks using multi-electrode arrays.

Efficient and safe use of hypothermia during various neuro-medical procedures requires sound understanding of low temperature effects on the neuronal network's activity. In this report, we introduce the use of cultivated dissociated neuronal networks on temperature controlled multi-electrode arrays (MEAs) as a simple methodology for studying the long-term effects of hypothermia. The networks exhibit spontaneous activity in the form of synchronized bursting events (SBEs), followed by long intervals of sporadic firing. Through the use of our correlation method, these SBEs can be clustered into sub-groups of similar spatio-temporal patterns. Application of hypothermia to the network resulted in a reduction in the SBE rate, the spike intensity and an increase in inter-neuronal correlations. Within 2h following the cessation of hypothermia, the cultured network returned to its initial spatio-temporal SBE structure. These results suggest that the network survived cold exposure and demonstrate the feasibility of long-term continuous neural network recording during hypothermic conditions.