10.1371/journal.pbio.1002217.g004

Conditional Pax6 expression in Tis21-positive APs changes the fate of the BP progeny.

Abstract

<p>Dorsolateral telencephalon of tamoxifen-treated E14.5 <i>Tis21</i>–CreER<sup>T2</sup> heterozygous mice electroporated at E13.5 with control (<b>A</b>,<b>C</b>,<b>D</b>,<b>F</b>,<b>H</b>–<b>J</b>) or Pax6-expressing (<b>B</b>,<b>C</b>,<b>E</b>,<b>F</b>–<b>J</b>) plasmid (see <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1002217#pbio.1002217.g002" target="_blank">Fig 2B</a>). (<b>A</b>,<b>B</b>,<b>D</b>,<b>E</b>) Tbr2 (<b>A</b>,<b>B</b>) or Sox2 (<b>D</b>,<b>E</b>) immunofluorescence (white) and RFP fluorescence (red), combined with DAPI staining (blue), on coronal 50-μm vibratome sections. Insets show representative examples of RFP-positive nuclei (outlined by dashed yellow lines) that are either Tbr2 (<b>A</b>,<b>B</b>) or Sox2 (<b>D</b>,<b>E</b>)-positive (yellow arrows, see also main panels) or-negative (yellow arrowheads). (<b>C</b>,<b>F</b>) Quantification of Tbr2- and RFP-positive cells (<b>C</b>) or Sox2- and RFP-positive cells (<b>F</b>) in the cortical wall (total), VZ, and SVZ, expressed as percentage of all RFP-positive cells in the cortical wall (200-μm wide area), upon control (Con, white) and Pax6 (black) electroporation. (<b>G</b>–<b>I</b>) Daughter cell pair analysis. (<b>G</b>) Examples of daughter cell pairs derived from Pax6-electroporated APs. Tbr2 immunofluorescence (white) and RFP fluorescence (red) on 12-μm cryosections. (Top) Tbr2+/Tbr2+, (middle) Tbr2+/Tbr2–, (bottom) Tbr2–/Tbr2–; yellow arrowheads, Tbr2–; yellow arrows, Tbr2+; yellow dashed lines, daughter cell pair nuclei. (<b>H</b>) Quantification of AP-derived daughter cell pairs upon control and Pax6 electroporation. Blue, Tbr2–/Tbr2–; red, Tbr2–/Tbr2+; green, Tbr2+/Tbr2+. Control, 23 pairs; Pax6, 24 pairs. (<b>I</b>) Distance of nuclei of the Tbr2–/Tbr2– daughter cell pairs from the ventricular surface upon control (Con, open circles, 7 pairs) and Pax6 (filled circles, 15 pairs) electroporation. Data indicate the position of the ventricular-most nucleus of each pair (see <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1002217#sec021" target="_blank">Materials and Methods</a>). Light and dark blue background indicates ventricular (within <27 μm from the ventricular surface) and abventricular (≥27 μm from the ventricular surface) location, respectively. (<b>J</b>) Number of nuclei in the VZ (200-μm-wide area) upon control (Con, white) and Pax6 (black) electroporation. (<b>A</b>,<b>B</b>,<b>D</b>,<b>E</b>,<b>G</b>) Dashed white lines, ventricular surface. Scale bars, 20 μm; inset scale bars, 5 μm. (<b>C</b>,<b>F</b>,<b>H</b>,<b>I</b>,<b>J</b>) Mean of three independent experiments, each being the average of two to four embryos. Error bars, SEM. * <i>p</i> <0.05, ** <i>p</i> <0.01.</p

Similar works

Full text

thumbnail-image
oai:figshare.com:article/1505608Last time updated on 2/12/2018

This paper was published in FigShare.

Having an issue?

Is data on this page outdated, violates copyrights or anything else? Report the problem now and we will take corresponding actions after reviewing your request.