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Cerebral Cortex Advance Access originally published online on November 21, 2008
Cerebral Cortex 2009 19(8):1723-1737; doi:10.1093/cercor/bhn194
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© The Author 2008. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org

Synaptogenesis in Purified Cortical Subplate Neurons

Claire E. McKellar1 and Carla J. Shatz2

Department of Neurobiology, Harvard Medical School, 220 Longwood Ave., Boston, MA 02110, USA, 1 Current address: HHMI/Janelia Farm Research Campus, 19700 Helix Dr., Ashburn, VA 20147, USA. Email: mckellarc{at}janelia.hhmi.org, 2 Current address: James H. Clark Center, 318 Campus Drive, W 1.1 Room W157, Stanford, CA 94305-5437, USA

Address correspondence to Claire McKellar, HHMI Janelia Farm Research Campus, 19700 Helix Drive, Ashburn, VA 20147, USA. Email: mckellarc{at}janelia.hhmi.org.

An ideal preparation for investigating events during synaptogenesis would be one in which synapses are sparse, but can be induced at will using a rapid, exogenous trigger. We describe a culture system of immunopurified subplate neurons in which synaptogenesis can be triggered, providing the first homogeneous culture of neocortical neurons for the investigation of synapse development. Synapses in immunopurified rat subplate neurons are sparse, and can be induced by a 48-h exposure to feeder layers of neurons and glia, an induction more rapid than any previously reported. Induced synapses are electrophysiologically functional and ultrastructurally normal. Microarray and real-time PCR experiments reveal a new program of gene expression accompanying synaptogenesis. Surprisingly few known synaptic genes are upregulated during the first 24 h of synaptogenesis; Gene Ontology annotation reveals a preferential upregulation of synaptic genes only at a later time. In situ hybridization confirms that some of the genes regulated in cultures are also expressed in the developing cortex. This culture system provides both a means of studying synapse formation in a homogeneous population of cortical neurons, and better synchronization of synaptogenesis, permitting the investigation of neuron-wide events following the triggering of synapse formation.

Key Words: culture • immunopurification • microarray • neocortex • synapse


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