Abstract
Forebrain embryonic zinc finger-like protein 2 (FEZF2) is a transcription factor involved in the embryonic development, differentiation and projection fates of neurons in the brain. It is expressed at high levels in corticospinal motor neurons and related subcerebral projection neurons during development, and this expression is retained in the adult brain. The expression of FEZF2 in progenitor cells, but only in a subset of mature neurons is of interest. The molecular network through which FEZF2 functions in the brain is still incomplete. This project primarily aimed to investigate a relationship between FEZF2 and the key neuronal microRNA miR-124 in a neural progenitor cell line and neurons.
To investigate the interaction between miR-124 and FEZF2, inducible FEZF2 knockdown cell lines were produced, and quantitative PCR was used to identify changes in FEZF2 related genes DPYSL3 and LEFTY1 and in miR-124 expression following FEZF2 knockdown. The expression of these genes was significantly altered in agreement with previous microarray data.
Primary neural cultures from transgenic mice expressing GFP under the regulation of the Fezf2 promoter were treated with miR inhibitors and the expression of Fezf2 analysed. Inhibition of miR-124 resulted in the reduction of Fezf2 expression.
The overexpression of FEZF2 required an FEZF2 expression construct. The full length FEZF2 coding sequence was amplified, and cloned into pEF1αIRES2GFP for future work.
These results demonstrate an interaction between miR-124 and FEZF2. Future experiments to determine whether this interaction is direct will provide insight into the mechanism of FEZF2 function in progenitor cells and neuronal differentiation.