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First published online 4 May 2005
doi: 10.1242/dev.01833

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Identification of Pax2-regulated genes by expression profiling of the mid-hindbrain organizer region

Maxime Bouchard^1,2,*,, David Grote^1,2,*, Sarah E. Craven³, Qiong Sun¹, Peter Steinlein¹ and Meinrad Busslinger¹

¹ Research Institute of Molecular Pathology, Vienna Biocenter, Dr Bohr-Gasse 7, 1030 Vienna, Austria
² McGill Cancer Centre, Biochemistry Department, McGill University, 3655 Promenade Sir-William-Osler, Montreal, Quebec, H3G 1Y6, Canada
³ Department of Molecular Biology, Genentech, 1 DNA Way, South San Francisco, CA 94080, USA

Author for correspondence (e-mail: maxime.bouchard{at}mcgill.ca)

Accepted 22 March 2005

The paired domain transcription factor Pax2 is required for the formation of the isthmic organizer (IsO) at the midbrain-hindbrain boundary, where it initiates expression of the IsO signal Fgf8. To gain further insight into the role of Pax2 in mid-hindbrain patterning, we searched for novel Pax2-regulated genes by cDNA microarray analysis of FACS-sorted GFP⁺ mid-hindbrain cells from wild-type and Pax2^–/– embryos carrying a Pax2^GFP BAC transgene. Here, we report the identification of five genes that depend on Pax2 function for their expression in the mid-hindbrain boundary region. These genes code for the transcription factors En2 and Brn1 (Pou3f3), the intracellular signaling modifiers Sef and Tapp1, and the non-coding RNA Ncrms. The Brn1 gene was further identified as a direct target of Pax2, as two functional Pax2-binding sites in the promoter and in an upstream regulatory element of Brn1 were essential for lacZ transgene expression at the mid-hindbrain boundary. Moreover, ectopic expression of a dominant-negative Brn1 protein in chick embryos implicated Brn1 in Fgf8 gene regulation. Together, these data defined novel functions of Pax2 in the establishment of distinct transcriptional programs and in the control of intracellular signaling during mid-hindbrain development.

Key words: Mid-hindbrain development, Pax2-regulated genes, Sef, Tapp1, Ncrms, En2, Brn1, Fgf8 regulation, Mouse

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