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doi: 10.1242/10.1242/dev.00337

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Targeted deletion of a branchial arch-specific enhancer reveals a role of dHAND in craniofacial development

Hiromi Yanagisawa¹, David E. Clouthier², James A. Richardson^1,3, Jeroen Charité^1,* and Eric N. Olson^1,

¹ Department of Molecular Biology, University of Texas Southwestern Medical Center at Dallas, TX 75390-9148, USA
³ Department of Pathology, University of Texas Southwestern Medical Center at Dallas, TX 75390-9148, USA
² Department of Molecular, Cellular and Craniofacial Biology, Birth Defects Center, School of Dentistry, University of Louisville, Louisville, KY 40292, USA
^* Present address: Department of Cell Biology and Genetics, Erasmus Medical Center Rotterdam, Dr Molewaterplein 50, 3015GE Rotterdam, The Netherlands

Author for correspondence (e-mail: eolson{at}hamon.swmed.edu)

Accepted 12 December 2002

The basic helix-loop-helix transcription factor dHAND is expressed in the mesenchyme of branchial arches and the developing heart. Mice homozygous for a dHAND (Hand2) null mutation die early in embryogenesis from cardiac abnormalities, precluding analysis of the potential role of dHAND in branchial arch development. Two independent enhancers control expression of dHAND in the heart and branchial arches. Endothelin-1 (ET-1) signaling regulates the branchial arch enhancer and is required for dHAND expression in the branchial arches. To determine the potential role of dHAND in branchial arch development and to assess the role of the ET-1-dependent enhancer in dHAND regulation in vivo, we deleted this enhancer by homologous recombination. Mice lacking the dHAND branchial arch enhancer died perinatally and exhibited a spectrum of craniofacial defects that included cleft palate, mandibular hypoplasia and cartilage malformations. Expression of dHAND was abolished in the ventolateral regions of the first and second branchial arches in these mutant mice, but expression was retained in a ventral domain where the related transcription factor eHAND is expressed. We conclude that dHAND plays an essential role in patterning and development of skeletal elements derived from the first and second branchial arches and that there are heterogeneous populations of cells in the branchial arches that rely on different cis-regulatory elements for activation of dHAND transcription.

Key words: dHand, Craniofacial development, Neural crest, Cleft palate

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