Impaired Embryonic Development in Mice Overexpressing the RNA-Binding Protein TIAR
Figure 5
In utero vs in vitro development of TIAR overexpressing embryos.
(A) Pre-implantation embryos from pregnant WT females mated with homozygous GFP-TIAR males were collected at various pre-implantation stages and observed for GFP expression. GFP expression starts at morula (mo) stage and persists at the early blastocyst stage (bl) where the blastocoele cavity starts to be clearly visible when observed upon bright field. (B) E3.5 embryos were collected from uterus of WT or PGK-Cre females mated with homozygous GFP-TIAR (TIAR) males, stained for TIAR and observed under confocal microscope using TOPRO to visualize the nucleus of each blastomere. The first column shows representative pictures of blastocysts while the second column shows blastomere magnification. IIary antibodies correspond to embryos for which the first step of labelling (anti-TIAR antibody) has been omitted to assess background fluoresence. (C) E3.5 embryos were collected from uterus of WT or PGK-Cre females mated with WT or homozygous GFP-TIAR (TIAR) males and the % of “healthy” blastocysts with no obvious fragmentation was counted. (D) Embryos from WT or PGK-Cre females mated with WT or homozygous GFP-TIA (TIAR) males were collected at E0.5 or E2.5, cultured in M16 medium until WT embryos hatched (for 2-4 days) and the % of “healthy” blastocysts with no obvious fragmentation was counted. n: number of counted embryos.