Bhlha9 regulates apical ectodermal ridge formation during limb development

J Bone Miner Metab. 2018 Jan;36(1):64-72. doi: 10.1007/s00774-017-0820-0. Epub 2017 Mar 21.


Split hand/foot malformation (SHFM) and SHFM combined with long-bone deficiency (SHFLD) are congenital dysgeneses of the limb. Although six different loci/mutations (SHFM1-SHFM6) have been found from studies on families with SHFM, the causes and associated pathogenic mechanisms for a large number of patients remain unidentified. On the basis of the identification of a duplicated gene region involving BHLHA9 in some affected families, BHLHA9 was identified as a novel SHFM/SHFLD-related gene. Although Bhlha9 is predicted to participate in limb development as a transcription factor, its precise function is unclear. Therefore, to study its physiological function, we generated a Bhlha9-knockout mouse and investigated gene expression and the associated phenotype in the limb bud. Bhlha9-knockout mice showed syndactyly and poliosis in the limb. Moreover, some apical ectodermal ridge (AER) formation related genes, including Trp63, exhibited an aberrant expression pattern in the limb bud of Bhlha9-knockout mice; TP63 (Trp63) was regulated by Bhlha9 on the basis of in vitro analysis. These observations suggest that Bhlha9 regulates AER formation during limb/finger development by regulating the expression of some AER-formation-related genes and abnormal expression of Bhlha9 leads to SHFM and SHFLD via dysregulation of AER formation and associated gene expression.

Keywords: Bhlha9; Split hand/foot malformation; Split hand/foot malformation combined with long-bone deficiency.

MeSH terms

  • Animals
  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Basic Helix-Loop-Helix Transcription Factors / metabolism*
  • Ectoderm / embryology*
  • Ectoderm / metabolism*
  • Extremities / embryology*
  • Fibroblast Growth Factor 8 / genetics
  • Fibroblast Growth Factor 8 / metabolism
  • Gene Expression Regulation, Developmental
  • HeLa Cells
  • Humans
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Phenotype
  • Phosphoproteins / metabolism
  • Trans-Activators / metabolism


  • BHLHA9 protein, human
  • Basic Helix-Loop-Helix Transcription Factors
  • Fgf8 protein, mouse
  • Phosphoproteins
  • Trans-Activators
  • Trp63 protein, mouse
  • bHLHA9 protein, mouse
  • Fibroblast Growth Factor 8