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Our previous transcriptome analysis identified 565 genes that are preferentially expressed in the developing brain of Ciona intestinalis larvae. Here, we show by in-situ hybridization that the spatial expression patterns of these brain-specific genes fall into different categories depending on the regions where the gene is expressed. For example, Ci-opsin3 and Ci-Dkk3 are expressed in the entire brain, Ci-tyrosinase and Ci-TYRP1 in the dorsal region, and Ci-synaptotagmin3, Ci-ZF399, and Ci-PTFb in the ventral region. Other genes are specific to the posterior, anterior, central, posterior and ventral, or anterior-ventral region of the brain. This regional expression of genes in the Ciona brain is not always associated with cell lineage, suggesting that complex mechanisms control the regionalized expression of brain-specific genes. [ABSTRACT FROM AUTHOR]