Abstract
Loss of synapses or alteration of synaptic activity is associated with cognitive impairment observed in a number of psychiatric and neurological disorders, such as schizophrenia and Alzheimer’s disease. Therefore successful development of in vitro methods that can investigate synaptic function in a high-throughput format could be highly impactful for neuroscience drug discovery. We present here the development, characterisation and validation of a novel high-throughput in vitro model for assessing neuronal function and synaptic transmission in primary rodent neurons. The novelty of our approach resides in the combination of the electrical field stimulation (EFS) with data acquisition in spatially separated areas of an interconnected neuronal network. We integrated our methodology with state of the art drug discovery instrumentation (FLIPR Tetra) and used selective tool compounds to perform a systematic pharmacological validation of the model. We investigated pharmacological modulators targeting pre- and post-synaptic receptors (AMPA, NMDA, GABA-A, mGluR2/3 receptors and Nav, Cav voltage-gated ion channels) and demonstrated the ability of our model to discriminate and measure synaptic transmission in cultured neuronal networks. Application of the model described here as an unbiased phenotypic screening approach will help with our long term goals of discovering novel therapeutic strategies for treating neurological disorders.
Highlights
Synapses are essential structural entities of neuronal tissue involved directly in the fast trans-neuronal transmission of information between different parts of the central and/or peripheral nervous system
This form of triggering neuronal activity which relies on induction of action potentials (APs) following application of an external electrical stimulus resembles the physiological form of neuronal activation
We focused on three different classes of drugs: i) compounds that modulate ion channels involved in neuronal excitability, ii) compounds that modulate post-synaptic receptors (AMPA, NMDA and GABA-A receptors) and iii) compounds that modulate pre-synaptic ion channels and receptors involved in neurotransmitter release, voltage gated calcium channels (VGCCs) and mGluR2/3 receptors
Summary
Synapses are essential structural entities of neuronal tissue involved directly in the fast trans-neuronal transmission of information between different parts of the central and/or peripheral nervous system They are involved in functional and structural plasticity within the complex neuronal circuits found in the brain. Whole cell patch-clamp can be used to record miniature post-synaptic excitatory (mEPSCs) or inhibitory (mIPSCs) currents to study indirectly synaptic function combined or not with synaptic plasticity[9]. Though this method doesn’t represent a direct measure of synaptic transmission it offers valuable insights into both pre- and postsynaptic mechanisms involved in synaptic transmission. We have used selective tool compounds and performed a systematic pharmacological validation of this model by investigating various ion channels and receptors known to be involved in neuronal excitability and synaptic transmission
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