Chimeric analysis of Notch2 function: a role for Notch2 in the development of the roof plate of the mouse brain

Dev Dyn. 2002 Oct;225(2):126-34. doi: 10.1002/dvdy.10140.

Abstract

Notch proteins are transmembrane receptors involved in cell-fate determination throughout development. Targeted disruption of either the Notch1 or Notch2 gene in mice results in embryonic lethality around embryonic day (E) 10.5 with widespread cell death. Although Notch1-deficient mice show disorganized somitogenesis, Notch2 mutants did not show definitive abnormalities in any tissue expressing high levels of the Notch2 gene, including the central nervous system. To study Notch2 function in development beyond the embryonic lethal stage, we performed chimeric analysis between Notch2 mutant and wild-type mouse embryos. Chimeric embryos developed normally and homozygous Notch2 mutant-specific cell death was not observed. Although chimeric embryos showed normal mosaicism until E9.5 in all tissues studied to date, Notch2 homozygous mutant cells failed to contribute to formation of the roof plate of the diencephalon and mesencephalon at later developmental stages, when Notch2 is normally expressed at high levels at there. Furthermore, Notch2 heterozygous mutant cells were also excluded from the roof plate of the chimera, however, Notch2 heterozygous mutant mice developed normally. We also showed that Wnt-1 and Mash1 expression patterns at the roof plate were disorganized in Notch2 homozygous mutant embryos. These results indicate that Notch2 plays an important role in development of the roof plate of the diencephalon and mesencephalon, and suggest that cellular rearrangement is involved in this process.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Basic Helix-Loop-Helix Transcription Factors
  • Cell Differentiation
  • DNA-Binding Proteins / biosynthesis
  • Diencephalon / embryology
  • Genotype
  • Green Fluorescent Proteins
  • Homozygote
  • In Situ Hybridization
  • Luminescent Proteins / metabolism
  • Mice
  • Mice, Transgenic
  • Microscopy, Fluorescence
  • Mutation
  • Proto-Oncogene Proteins / biosynthesis
  • RNA, Messenger / metabolism
  • Receptor, Notch2
  • Receptors, Cell Surface / genetics*
  • Receptors, Cell Surface / physiology*
  • Recombinant Fusion Proteins / metabolism
  • Signal Transduction
  • Time Factors
  • Transcription Factors / biosynthesis
  • Wnt Proteins
  • Wnt1 Protein
  • Zebrafish Proteins*
  • beta-Galactosidase / metabolism

Substances

  • Ascl1 protein, mouse
  • Basic Helix-Loop-Helix Transcription Factors
  • DNA-Binding Proteins
  • Luminescent Proteins
  • Notch2 protein, mouse
  • Proto-Oncogene Proteins
  • RNA, Messenger
  • Receptor, Notch2
  • Receptors, Cell Surface
  • Recombinant Fusion Proteins
  • Transcription Factors
  • Wnt Proteins
  • Wnt1 Protein
  • Wnt1 protein, mouse
  • Zebrafish Proteins
  • Green Fluorescent Proteins
  • beta-Galactosidase