Satb2Cre/+ mouse as a tool to investigate cell fate determination in the developing neocortex

J Neurosci Methods. 2017 Nov 1:291:113-121. doi: 10.1016/j.jneumeth.2017.07.023. Epub 2017 Aug 3.

Abstract

Background: Generation of different neuronal subtypes during neocortical development is the most important step in the establishment of cortical cytoarchitecture. The transcription factor Satb2 is expressed in neocortical projection neurons that send their axons intracortically as opposed to Satb2-negative neurons that preferentially project to subcortical targets.

New method: In this report, we present a novel method to carry out large scale screening for molecules that control cell fate in the developing neocortex. It is based on a Satb2Cre/+ mouse strain that expresses Cre recombinase from the Satb2 locus.

Results: By transfecting neuronould determine the proportion of cells that become al progenitors with a Cre-inducible reporter construct by nucleofection or in utero electroporation, we cSatb2-positive.

Comparison with existing methods: Compared to genetic tracing or lineage analysis, this method offers a fast, easy-to-perform, and reliable way of determining cell fate of newly born neurons.

Conclusions: We demonstrate that the Satb2Cre/+ mouse can be applied to study factors, such as small molecule inhibitors, sh-RNAs or overexpression constructs, that can alter the proportion of Satb2-positive cells and thus play key roles in differentiation and acquisition of cell fate.

Keywords: Cell fate acquisition; Cortical development; Neurogenesis; Satb2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation / physiology
  • Cells, Cultured
  • Electroporation
  • Flow Cytometry
  • Immunohistochemistry
  • Integrases / genetics
  • Integrases / metabolism
  • Matrix Attachment Region Binding Proteins / genetics
  • Matrix Attachment Region Binding Proteins / metabolism*
  • Mice, Transgenic
  • Models, Animal*
  • Neocortex / cytology*
  • Neocortex / growth & development*
  • Neocortex / metabolism
  • Neural Stem Cells / cytology
  • Neural Stem Cells / metabolism
  • Neurons / cytology*
  • Neurons / metabolism*
  • Repressor Proteins / metabolism
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*
  • Transfection
  • Tumor Suppressor Proteins / metabolism

Substances

  • Bcl11b protein, mouse
  • Matrix Attachment Region Binding Proteins
  • Repressor Proteins
  • SATB2 protein, mouse
  • Transcription Factors
  • Tumor Suppressor Proteins
  • Cre recombinase
  • Integrases