Cloning and expression analysis of the chick DAN gene, an antagonist of the BMP family of growth factors

Authors

  • Lisa M. Gerlach-Bank,

    1. Graduate Program in Cellular and Molecular Biology, University of Michigan Medical School, Ann Arbor, Michigan
    2. Center for Organogenesis, University of Michigan Medical School, Ann Arbor, Michigan
    Search for more papers by this author
  • Amanda D. Ellis,

    1. Department of Cell and Developmental Biology, University of Michigan Medical School, Ann Arbor, Michigan
    Search for more papers by this author
  • Bridgette Noonen,

    1. Department of Cell and Developmental Biology, University of Michigan Medical School, Ann Arbor, Michigan
    Search for more papers by this author
  • Kate F. Barald

    Corresponding author
    1. Graduate Program in Cellular and Molecular Biology, University of Michigan Medical School, Ann Arbor, Michigan
    2. Center for Organogenesis, University of Michigan Medical School, Ann Arbor, Michigan
    3. Department of Cell and Developmental Biology, University of Michigan Medical School, Ann Arbor, Michigan
    • University of Michigan, Department of Cell and Developmental Biology, 5740 Medical Science II, Box 0616, Ann Arbor, MI 48109-0616
    Search for more papers by this author

Abstract

Differential screening-selected gene aberrative in neuroblastoma (DAN) is a member of a cystine knot protein family that includes Cerberus and Gremlin. First isolated in a screen to identify genes down-regulated in transformed rat fibroblasts, DAN has subsequently been cloned in Xenopus, mouse, and human. Overexpression of DAN suppresses the transformed phenotype and retards the cell's entry into S phase. Biochemical analyses have demonstrated DAN's ability to bind bone morphogenetic proteins and antagonize their signaling activity. In this study, chick DAN was cloned and sequenced, revealing a conserved cystine knot region as well as an N-glycosylation site. A riboprobe was designed from the 3′ chick DAN coding sequence and used for analysis of DAN in the developing chick embryo by in situ hybridization. Chick DAN was expressed beginning at stage 10 in the developing somites and the medial otic epithelium. Expression in the neural layer of the eye became apparent at stage 14. By stage 17, expression had expanded to the base of the hindbrain. Limb bud labeling began at stage 20, whereas expression in the branchial arches appeared at stage 25. Chick DAN expression generally corresponded to that of mouse DAN expression as shown by comparative in situ hybridization. However, chick DAN was found in the otic epithelium and notochord, whereas mouse DAN was restricted to the overlying otic ectomesenchyme and was absent from the notochord. This observation suggests that DAN may play different roles in chick and mouse otic and notochord development. © 2002 Wiley-Liss, Inc.

Ancillary