As an approach toward understanding the molecular mechanisms of neuronal differentiation, we utilized DNA microarrays to elucidate global patterns of gene expression during pontocerebellar development. Through this analysis, we identified groups of genes specific to neuronal precursor cells, associated with axon outgrowth, and regulated in response to contact with synaptic target cells. In the cerebellum, we identified a phase of granule cell differentiation that is independent of interactions with other cerebellar cell types. Analysis of pontine gene expression revealed that distinct programs of gene expression, correlated with axon outgrowth and synapse formation, can be decoupled and are likely influenced by different cells in the cerebellar target environment. Our approach provides insight into the genetic programs underlying the differentiation of specific cell types in the pontocerebellar projection system.
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We are grateful to J. DeRisi and K. Burtis for use of their microarrayers and for help with microarray technology. S. Choksi, A. Finn, D. Lin, P. Luu, V. Peng, L. Simirenko, D. Speca, M. Szpara, and T. Alioto contributed to the microarray fabrication. We thank K. Vranizan for help with data analysis and K. Vranizan and E. Isacoff for comments on the manuscript. P.S. thanks B. Chih for help with data analysis and performing in situ hybridizations. This work was supported by grants to J.N., T.A.S., and T.P.S. from the National Institutes of Health and by funds from the Department of Molecular Cell Biology and Helen Wills Neuroscience Institute, University of California at Berkeley. P.S. is supported by awards from the Alfred P. Sloan Foundation and the Searle Scholars Program. E.D. was supported by a fellowship from the Helen Hay Whitney Foundation and a training grant from the National Institutes of Health.
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