Identification and characterization of regulators of neuronal differentiation during planarian regeneration

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Planarians are freshwater flatworms that have the remarkable talent to regrow lost body parts, even the head. This phenomenon is due to a high number of neoblasts, adult stem cells that can differentiate into any planarian cell type at the wound. Even though planarians are therefore an emerging model for stem cell biology, little is known about which signals specify neoblast fate and trigger their differentiation. The recent availability of a sequenced genome in combination with RNAi enables us now to study planarian gene function on a largescale basis. We plan to use RNAi in planarians to identify regulators and organizers of the regenerating nervous system. As cell-to-cell communication depends on secreted factors, we will generate an RNAi library against all secreted proteins encoded in the planarian genome. By systematic knockdown of these genes and subsequent analysis of the regenerating nervous system with neuron-specific antibodies, we hope to identify novel regulators of neuronal differentiation and growth. Novel proteins will be analyzed in secondary assays for their molecular functions, expression patterns, interaction partners, and their conservation and function in higher organisms such as frogs and Drosophila.