Figure 1

Neuronal progenitors can be purified directly from the rat telencephalon using flow cytometry. Dissociates were labeled using the pan-neuronal markers TnTx and ChTx in addition to A2B5 and Jones, which identify a separate lineage that also includes neuronal progenitors and later neuroglial progenitors (NGP). A. Bivariate FACS plots reveal a developmental increase in TnTx⁺/ChTx⁺/A2B5^-/Jones^- early neuronal progenitors (ENP) from 3% to 20.5% and the emergence of late neuronal progenitors (LNP) at E13. TnTx⁺/ChTx⁺ immunofluorescence signals were used to sort-purify populations of viable cells at embryonic days E11, E12 and E13. Sorting gates were set to collect TnTx⁺/ChTx⁺/A2B5^-/Jones^- ENPs and, at E13, LNPs, which were identified on the basis of relative fluorescence intensities of TnTx and ChTx staining reactions (upper left quadrant). B. Analysis of total RNA extracted from NPs isolated by FACS shows the presence of intact 18S and 28S rRNA bands (arrow heads). C. Principle component analysis demonstrates the reproducibility of the biological replicates. Red circles = E11 ENPs; yellow circles = E12 ENPs; light blue circles = E13 ENPs; dark blue circles = E13 LNPs. D. Scatter plot of two E11 ENP biological replicates. Correlation analysis was performed on the biological replicates within each group. The correlation coefficient was greater than 0.94 for each replicate.