Abstract

Genetic lineage tracing with electroporation is one of the most powerful techniques to target neural progenitor cells and their progeny. However, the spatiotemporal relationship between neural progenitors and their final phenotype remain poorly understood. One critical factor to analyze the cell fate of progeny is reporter integration into the genome of transfected cells. To address this issue, we performed postnatal and in utero co-electroporations of different fluorescent reporters to label, in both cerebral cortex and olfactory bulb, the progeny of subventricular zone neural progenitors. By comparing fluorescent reporter expression in the adult cell progeny, we show a differential expression pattern within the same cell lineage, depending on electroporation stage and cell identity. Further, while neuronal lineages arise from many progenitors in proliferative zones after few divisions, glial lineages come from fewer progenitors that accomplish many cell divisions. Together, these data provide a useful guide to select a strategy to track the cell fate of a specific cell population and to address whether a different proliferative origin might be correlated with functional heterogeneity.

Highlights

  • Neural cells are originated from a complex mix of distinct progenitors, which are pluripotent stem cells with restricted differentiation potential

  • Some radial glial cells (RGC) continue generating neurons and oligodendrocytes, while others RGCs transform into adult subventricular zone (SVZ) astrocytes, that differentiate to adult neural

  • Cell progeny showed three different patterns of reporter protein expression: mCherry positive cells, encoding the green fluorescent protein (EGFP) positive cells and cells expressing both reporters, depending on the uptake of one or both constructs into progenitors during electroporation

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Summary

Introduction

Neural cells are originated from a complex mix of distinct progenitors, which are pluripotent stem cells with restricted differentiation potential. NPCs undergo repeated symmetric divisions to self-renew while they divide asymmetrically to produce distinct neural and glial lineages at different time points. Those progenitors should maintain a balance between self-renewal and lineage commitment, becoming either neural stem cells or committed progenitors that undergo a limited number of divisions producing a lineage-restricted cell progeny. After an initial symmetrical amplification of the progenitor pool, radial glial cells (RGC) start to undergo asymmetrical divisions producing more restricted progenitors cells, including intermediate progenitor cells (see reviews by Götz and Huttner, 2005; Franco and Müller, 2013). Some RGCs continue generating neurons and oligodendrocytes, while others RGCs transform into adult subventricular zone (SVZ) astrocytes, that differentiate to adult neural

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