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First published online 7 January 2004
doi: 10.1242/dev.00948

Development 131, 613-627 (2004)
Published by The Company of Biologists 2004
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Prep1.1 has essential genetic functions in hindbrain development and cranial neural crest cell differentiation

Gianluca Deflorian1, Natascia Tiso1, Elisabetta Ferretti2, Dirk Meyer3, Francesco Blasi2, Marino Bortolussi1 and Francesco Argenton1,*

1 Dipartimento di Biologia, Universita' di Padova, Via U. Bassi 58/B, 35131 Padova, Italy
2 Department of Molecular Biology and Functional Genomics, Università Vita Salute San Raffaele and San Raffaele Scientific Institute, Via Olgettina, 58, 20132 Milano, Italy
3 Department of Developmental Biology, University of Freiburg, Haupstrasse 1, D-79104 Freiburg, Germany

* Author for correspondence (e-mail: francesco.argenton{at}unipd.it)

Accepted 27 October 2003

In this study we analysed the function of the Meinox gene prep1.1 during zebrafish development. Meinox proteins form heterotrimeric complexes with Hox and Pbx members, increasing the DNA binding specificity of Hox proteins in vitro and in vivo. However, a role for a specific Meinox protein in the regulation of Hox activity in vivo has not been demonstrated. In situ hybridization showed that prep1.1 is expressed maternally and ubiquitously up to 24 hours post-fertilization (hpf), and restricted to the head from 48 hpf onwards. Morpholino-induced prep1.1 loss-of-function caused significant apoptosis in the CNS. Hindbrain segmentation and patterning was affected severely, as revealed by either loss or defective expression of several hindbrain markers (foxb1.2/mariposa, krox20, pax2.1 and pax6.1), including anteriorly expressed Hox genes (hoxb1a, hoxa2 and hoxb2), the impaired migration of facial nerve motor neurons, and the lack of reticulospinal neurons (RSNs) except Mauthner cells. Furthermore, the heads of prep1.1 morphants lacked all pharyngeal cartilages. This was not caused by the absence of neural crest cells or their impaired migration into the pharyngeal arches, as shown by expression of dlx2 and snail1, but by the inability of these cells to differentiate into chondroblasts. Our results indicate that prep1.1 has a unique genetic function in craniofacial chondrogenesis and, acting as a member of Meinox-Pbc-Hox trimers, it plays an essential role in hindbrain development.

Key words: Vertebrate, Zebrafish, Hox, Pbx, Meis, Prep, Rhombomere, Segmentation, Neural Crest, Pharingeal arch, Morpholino




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