Submitted on June 15, 2004
Accepted on March 24, 2005
Gene expression profiling of cerebellar development with high-throughput functional analysis
Sakae Saito1, Kimi Honma2, Hiroko Kita-Matsuo1, Takahiro Ochiya3, and Kikuya Kato4*
1 Taisho Laboratory of Functional Genomics, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0101, Japan
2 Koken Bioscience Institute, Tokyo 115-0051, Japan
3 National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan
4 Research Institute, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-2 Nakamichi, Higashinari-ku, Osaka 537-8511, Japan
* To whom correspondence should be addressed. E-mail: katou-ki{at}mc.pref.osaka.jp.
We measured the expression levels of 450 genes during mouse postnatal cerebellar development by quantitative PCR using RNA purified from layers of the cerebellar cortex. Principal component analysis of the data matrix demonstrated that the first and second components correspond to general levels of gene expression and gene
expression patterns, respectively. We introduced 288 of the 450 genes into PC12 cells using a high-throughput transfection assay based on atelocollagen, and determined the ability of each gene to promote neurite outgrowth or cell proliferation. Five genes induced neurite outgrowth and seven genes enhanced proliferation. Evaluation of the
functional data and gene expression patterns showed that none of these genes exhibited elevated expression at maturation, suggesting that genes characteristic of mature
neurons are not likely to participate in neuronal development. These results demonstrate that functional data can facilitate interpretation of expression profiles and identification of new molecules that participate in biological processes.
