Publication date: Available online 3 August 2017
Source:Journal of Neuroscience Methods
Author(s): Mateusz Cyryl Ambrozkiewicz, Paraskevi Bessa, Andrea Salazar-Lázaro, Valentina Salina, Victor Tarabykin
BackgroundGeneration 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 MethodIn 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.ResultsBy transfecting neuronal progenitors with a Cre-inducible reporter construct by nucleofection or in utero electroporation, we could determine the proportion of cells that become Satb2-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.ConclusionsWe 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.
Graphical abstract
from #ORL-AlexandrosSfakianakis via ola Kala on Inoreader http://ift.tt/2huKBtx
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