%0 Journal Article
%T Lack of the brain-specific isoform of apoptosis-inducing factor aggravates cerebral damage in a model of neonatal hypoxia¨Cischemia
%A Anders St£¿hlberg
%A Carina Mallard
%A Changlian Zhu
%A Cuicui Xie
%A Daniel Andersson
%A Guido Kroemer
%A Henrik Hagberg
%A Juan Rodriguez
%A Kai Zhou
%A Kaiming Huo
%A Klas Blomgren
%A Nazanine Modjtahedi
%A Qinghe Xing
%A Tao Li
%A Xiaoyang Wang
%A Yafeng Wang
%A Yanyan Sun
%A Yaodong Zhang
%A Yiran Xu
%J Archive of "Cell Death & Disease".
%D 2019
%R 10.1038/s41419-018-1250-1
%X a The Aif2 genome locus (WT allele) was modified in embryonic stem cells using the targeting vector. LoxP recombination sites (red bars) along with an FRT (green bar)-flanked neomycin-selection cassette were introduced flanking exon 2b (flox allele). The expression of FLP recombinase results in the removal of the neomycin cassette (flox delta-neo allele). Expression of the Cre recombinase resulted in the excision of exon 2b (KO Allele) and a frameshift, generating a null allele. FRT flippase recognition target sites, FLP flippase. Primer pair P1/P2 distinguishes WT from flox delta neo; primer pair P3/P2 identifies the flox allele; primer pair P4/P2 distinguishes WT from KO. b The relative mRNA expression showed that there was a significant increase in Aif1 mRNA when the Aif2 mRNA was knocked out in P60 adult mice compared to WT littermates (1.19£¿¡À£¿0.07 in P9 vs. 2.26£¿¡À£¿0.17 in P60 Aif2 KO; **p£¿<£¿0.01) (P9 WT: n£¿=£¿7; P9 Aif2 KO: n£¿=£¿8; P60 WT: n£¿=£¿4; P60 Aif2 KO: n£¿=£¿10). c There was a significant increase in the relative expression of Aif2 mRNA in WT mice at P9 and P60 (1.00£¿¡À£¿0.12 in P9 vs. 1.64£¿¡À£¿0.13 in P60; *p£¿<£¿0.05) (P9: n£¿=£¿7; P60: n£¿=£¿4). d Representative western blot analysis of AIF, CHCHD4, cytochrome C (CYT C), SOD2, HSP70, and complex-I 39£¿kDda subunit (COX I) in both WT and KO mice at P9 under physiological conditions. e Representative Ki67 immunostaining in the dentate gyrus of the hippocampus at P60 and quantification of Ki67-positive cells showing no difference between WT and KO. f DCX staining in the dentate gyrus. The number of DCX-positive cells was 27% higher in the KO mice (n£¿=£¿8) compared to WT mice (n£¿=£¿13). g The photomicrographs show representative ovarian follicles for both WT and KO mice at P60 (n£¿=£¿4 for WT and n£¿=£¿6 for KO). h The total number of pups over time, starting from 10 weeks, for WT and KO mice. i The average number of pups per litter in WT (litters£¿=£¿5) and KO mice (litters£¿=£¿4
%U https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6315035/