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. 2016 Jul 1;143(13):2356–2366. doi: 10.1242/dev.135913

Fig. 1.

Fig. 1.

Apclox/lox;P0-Cre mice exhibit hindlimb weakness and impaired axonal conduction in the sciatic nerve. (A) Excision of Apc exon 14 was verified in sciatic nerves of P7 Apclox/lox;P0-Cre mice using genomic DNA and primers that recognize the excised allele only: both the Apclox/lox;P0-Cre and Apclox/lox (used as control) carry the Apc exon 14 loxP-flanked allele (top panel), whereas the excised allele can be detected in sciatic nerve of Apclox/lox;P0-Cre mice only (bottom panel). (B) The excised Apc allele was also detected at different time points by quantitative PCR analysis on genomic DNA extracted from sciatic nerves of Apclox/lox;P0-Cre and Apclox/lox (control) mice. Il2 was used as an internal control gene for normalization. (C) In order to study the recombination efficiency of the P0-Cre line, we generated a ROSA26-stop-EYFP;P0-Cre reporter line. YFP+ cells were labeled in sciatic nerves of ROSA26-stop-EYFP;P0-Cre mice at different ages with an anti-GFP antibody and the percentage of DAPI+ cells expressing the YPF reporter gene protein was quantified. (D) Apclox/lox;P0-Cre mice exhibit hindlimb clenching when suspended by the tail, a common neuropathic phenotype. (E) Apclox/lox;P0-Cre mice also suffer from hindlimb weakness as detected by the grip strength assay. (F) The conduction velocity and the amplitude of compound muscle action potential were both significantly reduced in sciatic nerves of the Apclox/lox;P0-Cre mice. For detection of the excised allele, three to four animals were taken per each time point. The excised allele was not detected in the Apclox/lox mice at any time point. For reporter line assay, sciatic nerves from three to four animals were harvested per each time point. No YFP-positive cells were detected in control littermate ROSA26-stop-EYFP mice (Cre negative) at any time point. For grip strength assay: Apclox/lox, n=11 mice and Apclox/lox;P0-Cre, n=16 mice at P60. For electrophysiological studies: Apclox/lox, n=7 mice and Apclox/lox;P0-Cre, n=6 mice at P60. DL, distal latency; CV, conduction velocity; dAmp, distal amplitude. *P<0.05, **P<0.001.