Abstract
Numerous risk genes have recently been implicated in susceptibility to autism and schizophrenia. Translating such genetic findings into disease-relevant neurobiological mechanisms is challenging due to the lack of throughput assays that can be used to assess their functions on an appropriate scale. To address this issue, we explored the feasibility of using a micro-electrode array (MEA) as a potentially scalable assay to identify the electrical network phenotypes associated with risk genes. We first characterized local and global network firing in cortical neurons with MEAs, and then developed methods to analyze the alternation between the network active period (NAP) and the network inactive period (NIP), each of which lasts tens of seconds. We then evaluated the electric phenotypes of neurons derived from Shank3 knockout (KO) mice. Cortical neurons cultured on MEAs displayed a rich repertoire of spontaneous firing, and Shank3 deletion led to reduced firing activity. Enhancing excitation with CX546 rescued the deficit in the spike rate in the Shank3 KO network. In addition, the Shank3 KO network produced a shorter NIP, and this altered network firing pattern was normalized by clonazepam, a positive modulator of the GABAA receptor. MEA recordings revealed electric phenotypes that displayed altered excitation and inhibition in the network lacking Shank3. Thus, our study highlights MEAs as an experimental framework for measuring multiple robust neurobiological end points in dynamic networks and as an assay system that could be used to identify electric phenotypes in cultured neuronal networks and to analyze additional risk genes identified in psychiatric genetics.
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Acknowledgements
We thank Dr Stefano Vicini from Georgetown University for helpful discussions. We thank Caroline Maguire, Gregory Foos and Triana Dalia for technical support. We thank the members of the Feng Lab and the Pan Group for helpful discussions. This work was supported by National Institute of Health (R21MH099448 to JQP and R01MH097104 to GF), and by the Stanley Center for Psychiatric Research at the Broad Institute of MIT and Harvard, a grant from the Fidelity Bioscience Research Initiative, and the Poitras Center for Affective Disorders Research at the McGovern Institute for Brain Research at MIT.
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Lu, C., Chen, Q., Zhou, T. et al. Micro-electrode array recordings reveal reductions in both excitation and inhibition in cultured cortical neuron networks lacking Shank3. Mol Psychiatry 21, 159–168 (2016). https://doi.org/10.1038/mp.2015.173
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DOI: https://doi.org/10.1038/mp.2015.173
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