Real-time PCR quantification of gene expression in embryonic mouse tissue

Methods Mol Biol. 2014:1092:81-94. doi: 10.1007/978-1-60327-292-6_6.

Abstract

The Gbx family of transcription factors consists of two closely related proteins GBX1 and GBX2. A defining feature of the GBX family is a highly conserved 60 amino acid DNA-binding domain, which differs by just two amino acids. Gbx1 and Gbx2 are co-expressed in several areas of the developing central nervous system including the forebrain, anterior hindbrain, and spinal cord, suggesting the potential for genetic redundancy. However, there is a spatiotemporal difference in expression of Gbx1 and Gbx2 in the forebrain and spinal cord. Gbx2 has been shown to play a critical role in positioning the midbrain/hindbrain boundary and developing anterior hindbrain, whereas gene-targeting experiments in mice have revealed an essential function for Gbx1 in the spinal cord for normal locomotion. To determine if Gbx2 could potentially compensate for a loss of Gbx1 in the developing spinal cord, we performed real-time PCR to examine levels of Gbx2 expression in Gbx1(-/-) spinal cord at embryonic day (E) 13.5, a developmental stage when Gbx2 is rapidly downregulated. We demonstrate that Gbx2 expression is elevated in the spinal cord of Gbx1(-/-) embryos.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Gene Expression Regulation, Developmental*
  • Homeodomain Proteins / biosynthesis*
  • Homeodomain Proteins / genetics
  • Mice
  • Molecular Biology / methods
  • Prosencephalon / embryology
  • Rhombencephalon / embryology
  • Spinal Cord / embryology
  • Tissue Distribution

Substances

  • Gbx1 protein, mouse
  • Gbx2 protein, mouse
  • Homeodomain Proteins