Fig. 4

Effects of in utero PGF overexpression on fetal growth and cortical vasculature during prenatal alcohol exposure. (a, b) PGF CRISPR-dCas 9 activation approach coupled with in utero electroporation of the placenta was done at GD13 (a) and overexpression of PLGF controlled at GD20 (b). In the alcohol group, in utero exposure occurred from GD15 to GD20. (c, d) Visualization of E20 fetuses from pregnant mice exposed to NaCl (c) or alcohol (d). Note the small size of alcohol-exposed fetuses. Green bars indicate morphometric measures that have been done (head size (a); body size (b); abdomen size (c) and whole fetus size (a + b). (e, f) Visualization of E20 fetuses after in utero electroporation of PGF CRISPR-dCas9 plasmids in placentae from control (e) or alcohol-exposed pregnant mice (f). (g, h) Quantification of abdomen (g) and whole fetus (h) sizes in control (NaCl) and alcohol groups. In a same uterine horn some placentae were not electroporated (black bars), electroporated with control CRISPR-Cas9 plasmids (grey bars) or electroporated with PGF CRISPR-dCas9 plasmids (white bars). ##p < 0.01; ###p < 0.001; ####p < 0.0001 vs the control group and *p < 0.05; **p < 0.01; ****p < 0.0001 as indicated using the two way ANOVA test followed by Tukey’s post hoc test. (i-k) Visualization of the vasculature in the cortex of E20 fetuses from control (NaCl)/non-transfected (i), alcohol/control CRISPR-Cas9 transfected (j) and alcohol/ PGF CRISPR-dCas9 transfected (k) placentae. (l) Quantification of the percentage of radial vessels in the cortex of E20 fetuses from not electroporated (black bars), electroporated with control CRISPR-Cas9 plasmids (grey bars) and electroporated with PGF CRISPR-dCas9 plasmids (white bars) placentae. #p < 0.05 vs the control group and *p < 0.05 as indicated using the two way ANOVA test followed by Tukey’s post hoc test