Construction and characterization of a sox9b transgenic reporter line

Int J Dev Biol. 2014;58(9):693-9. doi: 10.1387/ijdb.140288jp.

Abstract

The transcription factor SOX9 is a member of the SRY-related high-mobility-group box (SOX) superfamily of genes. In mammals, Sox9 plays important roles in many developmental processes including craniofacial, skeletal and heart morphogenesis, retinal and brain development, and gonad differentiation. Human mutations in SOX9 or the SOX9 promoter result in campomelic dysplasia, a severe genetic disorder, which disrupts skeletal, craniofacial, cardiac, neural and reproductive development. Due to the duplication of the teleost fish genome, zebrafish (Danio rerio) have two Sox9 genes: sox9a and sox9b. Loss of sox9b in zebrafish results in loss of function phenotypes that are similar to those observed in humans and mice. In order to generate a transgenic sox9b:EGFP reporter line, we cloned a 2450 bp fragment of the sox9b promoter and fused it to an EGFP reporter. Consistent with reported sox9b expression and function, we observed sox9b:EGFP in the developing heart, skeletal and craniofacial structures, brain, retina, and ovaries. Our resulting transgenic line is a useful tool for identifying and studying sox9b function in development and visualizing a number of zebrafish organs and tissues in which sox9b is normally expressed.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified / genetics
  • Animals, Genetically Modified / growth & development*
  • Animals, Genetically Modified / metabolism
  • Brain / embryology
  • Brain / metabolism
  • Craniofacial Abnormalities / embryology
  • Craniofacial Abnormalities / metabolism
  • Embryo, Nonmammalian / cytology*
  • Embryo, Nonmammalian / metabolism*
  • Female
  • Gene Expression Regulation, Developmental*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism*
  • Heart / embryology
  • Heart / physiology
  • Humans
  • Immunoenzyme Techniques
  • In Situ Hybridization
  • Mice
  • Muscle, Skeletal / embryology
  • Muscle, Skeletal / metabolism
  • Ovary / embryology
  • Ovary / metabolism
  • Retina / embryology
  • Retina / metabolism
  • SOX9 Transcription Factor / genetics
  • SOX9 Transcription Factor / metabolism*
  • Zebrafish / genetics
  • Zebrafish / growth & development*
  • Zebrafish / metabolism
  • Zebrafish Proteins / genetics
  • Zebrafish Proteins / metabolism*

Substances

  • SOX9 Transcription Factor
  • Sox9b protein, zebrafish
  • Zebrafish Proteins
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins