Synaptic signal streams generated by ex vivo neuronal networks contain non-random, complex patterns.

Cultured embryonic neurons develop functional networks that transmit synaptic signals over multiple sequentially connected neurons as revealed by multi-electrode arrays (MEAs) embedded within the culture dish. Signal streams of ex vivo networks contain spikes and bursts of varying amplitude and duration. Despite the random interactions inherent in dissociated cultures, neurons are capable of establishing functional ex vivo networks that transmit signals among synaptically connected neurons, undergo developmental maturation, and respond to exogenous stimulation by alterations in signal patterns.

Acceleration of myofiber formation in culture by a digitized synaptic signal.

Developing myofibers require chemical and electrical stimulation to induce functional muscle tissue. Tissue engineering protocols utilize either or both of these to initiate differentiation ex vivo. Current methodologies typically deliver multi-volt electrical signals, which may be hazardous to developing tissues.

Critical role for inhibitory neurons in modulation of synaptic signaling in ex vivo neuronal networks.

A number of laboratories have modeled aspects of synaptic plasticity using neuronal networks established on micro-electrode arrays. Such studies demonstrate that external stimulation can increase or hasten maturation of network signaling as evidenced an increase in complex bursts. Herein, we demonstrate that repetitive stimulation with a recorded synaptic signal was capable of increasing overall signaling, including the percentage of bursts, over a 5-day period, but that this increase was completely prevented by the presence of the GABAergic antagonist bicuculline.

Transient epileptiform signaling during neuronal network development: regulation by external stimulation and bimodal GABAergic activity.

A predominance of excitatory activity, with protracted appearance of inhibitory activity, accompanies cortical neuronal development. It is unclear whether or not inhibitory neuronal activity is solicited exclusively by excitatory neurons or whether the transient excitatory activity displayed by developing GABAergic neurons contributes to an excitatory threshold that fosters their conversion to inhibitory activity. We addressed this possibility by culturing murine embryonic neurons on multi-electrode arrays.

Accelerated establishment of mature signaling patterns following stimulation of developing neuronal networks: "learning" versus "plasticity".

Neuronal networks established on micro-electrode arrays provide useful models for synaptic plasticity. Whether or not this represents a facet of learning is debated since ex vivo networks are deprived of organismal interaction with the environment. We compared developmental signaling of such networks with and without stimulation with a prerecorded synaptic signal from another mature culture as a model of sensory input.

Stimulation with a low-amplitude, digitized synaptic signal to invoke robust activity within neuronal networks on multielectrode arrays.

Multielectrode arrays (MEAs) are used for analysis of neuronal activity. Here we report two variations on commonly accepted techniques that increase the precision of extracellular electrical stimulation: (i) the use of a low-amplitude recorded spontaneous synaptic signal as a stimulus waveform and (ii) the use of a specific electrode within the array adjacent to the stimulus electrode as a hard-grounded stimulus signal return path. Both modifications remained compatible with manipulation of neuronal networks.