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Spatiotemporal analyses of neural lineages after embryonic and postnatal progenitor targeting combining different reporters.

Figueres-Oñate M, García-Marqués J, Pedraza M, De Carlos JA, López-Mascaraque L - Front Neurosci (2015)

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Instituto Cajal-Consejo Superior de Investigaciones Científicas, Department of Molecular, Cellular and Developmental Neurobiology Madrid, Spain.

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.

No MeSH data available.


Adult glial transfected cells after postnatal electroporation (P1) (A) Electroporation area showing targeted cells located in the dorso-lateral wall of the lateral ventricle. (B–D) Glial cells derived from SVZ progenitors expressed just the integrated construct (green) whereas remaining SVZ-cells highly co-expressed both constructs (yellow). Transfected cells with glial morphology located in the withe matter (B), in lower cortical layers (C), and in the striatum (D). Typical morphologies of astrocytes (D, asterisk) and oligodendrocytes (D, arrowhead). CC, corpus callosum; LV, Lateral ventricle; Str, striatum.
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Figure 5: Adult glial transfected cells after postnatal electroporation (P1) (A) Electroporation area showing targeted cells located in the dorso-lateral wall of the lateral ventricle. (B–D) Glial cells derived from SVZ progenitors expressed just the integrated construct (green) whereas remaining SVZ-cells highly co-expressed both constructs (yellow). Transfected cells with glial morphology located in the withe matter (B), in lower cortical layers (C), and in the striatum (D). Typical morphologies of astrocytes (D, asterisk) and oligodendrocytes (D, arrowhead). CC, corpus callosum; LV, Lateral ventricle; Str, striatum.

Mentions: Electroporation of postnatal SVZ progenitors yielded glial cells after several weeks (Figure 5). Interestingly, those glial cells, generated from postnatal precursors, expressed only the integrable construct at P30. After postnatal electroporation into the dorsolateral part of the neonatal SVZ (Figure 5A), glial cells were found in the white matter (Figure 5B) and within the gray matter both in lower cortical layers (Figure 5C) and the striatum (Figure 5D). Consequently those glial cells probably arose from SVZ progenitors after a long number of divisions.


Spatiotemporal analyses of neural lineages after embryonic and postnatal progenitor targeting combining different reporters.

Figueres-Oñate M, García-Marqués J, Pedraza M, De Carlos JA, López-Mascaraque L - Front Neurosci (2015)

Adult glial transfected cells after postnatal electroporation (P1) (A) Electroporation area showing targeted cells located in the dorso-lateral wall of the lateral ventricle. (B–D) Glial cells derived from SVZ progenitors expressed just the integrated construct (green) whereas remaining SVZ-cells highly co-expressed both constructs (yellow). Transfected cells with glial morphology located in the withe matter (B), in lower cortical layers (C), and in the striatum (D). Typical morphologies of astrocytes (D, asterisk) and oligodendrocytes (D, arrowhead). CC, corpus callosum; LV, Lateral ventricle; Str, striatum.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4362314&req=5

Figure 5: Adult glial transfected cells after postnatal electroporation (P1) (A) Electroporation area showing targeted cells located in the dorso-lateral wall of the lateral ventricle. (B–D) Glial cells derived from SVZ progenitors expressed just the integrated construct (green) whereas remaining SVZ-cells highly co-expressed both constructs (yellow). Transfected cells with glial morphology located in the withe matter (B), in lower cortical layers (C), and in the striatum (D). Typical morphologies of astrocytes (D, asterisk) and oligodendrocytes (D, arrowhead). CC, corpus callosum; LV, Lateral ventricle; Str, striatum.
Mentions: Electroporation of postnatal SVZ progenitors yielded glial cells after several weeks (Figure 5). Interestingly, those glial cells, generated from postnatal precursors, expressed only the integrable construct at P30. After postnatal electroporation into the dorsolateral part of the neonatal SVZ (Figure 5A), glial cells were found in the white matter (Figure 5B) and within the gray matter both in lower cortical layers (Figure 5C) and the striatum (Figure 5D). Consequently those glial cells probably arose from SVZ progenitors after a long number of divisions.

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Instituto Cajal-Consejo Superior de Investigaciones Científicas, Department of Molecular, Cellular and Developmental Neurobiology Madrid, Spain.

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.

No MeSH data available.