1. Targeted disruption of the mouse Pex14 gene.
Targeted disruption of the mouse Pex14 gene. (A) Schematic representation of the Pex14 genome locus (top), targeting vector (pMC-KO, middle), and targeted allele of the mutated locus following the homologous recombination (bottom). Exon sequences are indicated by black bars and boxes. (B) PCR-based genotyping using tail-derived DNA of wild-type (+/+), heterozygous (+/ΔC), and homozygous (ΔC/ΔC) Pex14 mutant mice. Arrows indicate the sequences used for primers described in the Materials and Methods section. Primers P14F (F) and P14R (R) amplify a 599-bp fragment, and primers F and KN52-2 (K) amplify a 169-bp fragment specific for the recombined Pex14 gene. (C) Schematic structure of predicted Pex14p proteins in wild-type mice, Pex14 mutant mice (Pex14p-Mut), and patients with a Pex14 nonsense mutation, C553T (Pex14p-Q185X). Gray bar, Pex5p-binding domain; brown bar, transmembrane domain (TM); yellow bar, coiled-coil domain (CC); shaded area, altered amino acid sequence caused by a frameshift mutation (129–163). (D) Phenotypic appearance of Pex14 mutant mice ∼12 h after birth. Scale bar, 1 cm. (E) Postnatal body weights were determined at P0.5. The number of pups with each genotype is indicated. ***P < 0.001, by Dunnett's test compared with +/+. (F) Cresyl violet staining of a coronal section of the cortex from a P0.5 mouse. The open arrows indicate accumulated neurons in the intermediate zone (IZ). Scale bar, 100 μm. (G) Enlarged view of the boxed regions in F. Cortical layers are indicated on the left. The boundaries of the cortical plates are indicated by dashed lines. Scale bar, 50 μm. CP, cortical plate; DT-A, diphtheria toxin A cassette; Neor, Neomycin-resistant gene; VZ, ventricular zone.
Yuichi Abe et al. LSA 2018;1:e
© 2018 Abe et al.