ABSTRACT
In this study, we demonstrate capabilities to selectively manipulate dissociated co-cultures of neurons plated in dual-compartment devices. Synaptic receptor antagonists and tetrodotoxin solutions were used to selectively control and study the network-wide burst propagation and cell firing in cortical-cortical and cortical-thalamic co-culture systems. The results show that in cortical-thalamic dissociated co-cultures, burst events initiate in the cortical region and propagate to the thalamic region and the burst events in thalamic region can be controlled by blocking the synaptic receptors in the cortical region. Whereas, in cortical-cortical co-culture system, one of the region acts as a site of burst initiation and facilitate propagation of bursts in the entire network. Tetrodotoxin, a sodium channel blocker, when applied to either of the regions blocks the firing of neurons in that particular region with significant influence on the firing of neurons in the other region. The results demonstrate selective pharmacological manipulation capabilities of co-cultures in a dual compartment device and helps understand the effects of neuroactive compounds on networks derived from specific CNS tissues and the dynamic interaction between them.
Subject(s)
Cerebral Cortex/cytology , Coculture Techniques/instrumentation , Microfluidic Analytical Techniques , Neurons/drug effects , Thalamus/cytology , Action Potentials/drug effects , Animals , Cell Communication/drug effects , Cerebral Cortex/embryology , Dimethylpolysiloxanes , Equipment Design , Excitatory Amino Acid Antagonists/pharmacology , GABA Antagonists/pharmacology , Genes, Reporter , Green Fluorescent Proteins/analysis , Green Fluorescent Proteins/genetics , Microelectrodes , Nerve Net/physiology , Neurites/ultrastructure , Neurons/cytology , Neurotoxins/pharmacology , Nylons , Primary Cell Culture/methods , Rats , Rats, Wistar , Receptors, Neurotransmitter/drug effects , Receptors, Neurotransmitter/physiology , Sodium Channel Blockers/pharmacology , Tetrodotoxin/pharmacology , Thalamus/embryologyABSTRACT
Co-cultures containing dissociated cortical and thalamic cells may provide a unique model for understanding the pathophysiology in the respective neuronal sub-circuitry. In addition, developing an in vitro dissociated co-culture model offers the possibility of studying the system without influence from other neuronal sub-populations. Here we demonstrate a dual compartment system coupled to microelectrode arrays (MEAs) for co-culturing and recording spontaneous activities from neuronal sub-populations. Propagation of electrical activities between cortical and thalamic regions and their interdependence in connectivity is verified by means of a cross-correlation algorithm. We found that burst events originate in the cortical region and drive the entire cortical-thalamic network bursting behavior while mutually weak thalamic connections play a relevant role in sustaining longer burst events in cortical cells. To support these experimental findings, a neuronal network model was developed and used to investigate the interplay between network dynamics and connectivity in the cortical-thalamic system.
