The midbrain-hindbrain boundary (MHB) is a long-lasting organizing center in the vertebrate neural tube that is both necessary and sufficient for the ordered development of midbrain and anterior hindbrain (midbrain-hindbrain domain, MH). The MHB also coincides with a pool of progenitor cells that contributes neurons to the entire MH. Here we show that the organizing activity and progenitor state of the MHB are co-regulated by a single microRNA, miR-9, during late embryonic development in zebrafish. Endogenous miR-9 expression, initiated at late stages, selectively spares the MHB. Gain- and loss-of-function studies, in silico predictions and sensor assays in vivo demonstrate that miR-9 targets several components of the Fgf signaling pathway, thereby delimiting the organizing activity of the MHB. In addition, miR-9 promotes progression of neurogenesis in the MH, defining the MHB progenitor pool. Together, these findings highlight a previously unknown mechanism by which a single microRNA fine-tunes late MHB coherence via its co-regulation of patterning activities and neurogenesis.
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We are grateful to members of the L.B.-C. laboratory for discussions and to M. Götz, W. Norton and M. Wassef for their insightful ideas and critical reading of the manuscript. This work was funded by a junior group grant from the Volkswagen Association, the EU 6th framework integrated project ZF-Models (contract No. LSHC-CT-2003-503466), the Life Science Association (No. GSF 2005/01), a special research grant from the Institut du Cerveau et de la Moelle épinière, the Excellence Center for Protein Science, Munich, and the Helmholtz 'Impuls und Vernetzungsfond'.
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Leucht, C., Stigloher, C., Wizenmann, A. et al. MicroRNA-9 directs late organizer activity of the midbrain-hindbrain boundary. Nat Neurosci 11, 641–648 (2008) doi:10.1038/nn.2115
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