Timed data for onset and survival.

<p>Given are parameters for disease onset (A-C), and survival (D) for the SOD-1 (G93A) (n = 17) and SOD-1 (G93A)^PS mice (n = 8). (A) Onset of weight decrease was defined as a drop of 5% of the mouse maximal weight. (B) First manifestation of symptoms that are linked to gait impairment in terms of limbs coordination and overall stance, and (C) time to first manifestation of paresis. (D) End point of the disease, defined as the inability of the mouse to right itself within 30 seconds.</p

Listed are key characteristics of ALS-like symptoms in the original SOD-1 (G93A) and in the new SOD-1 (G93A)^PS line (mean±SEM).

<p>Listed are key characteristics of ALS-like symptoms in the original SOD-1 (G93A) and in the new SOD-1 (G93A)^PS line (mean±SEM).</p

Weight loss and muscular strength deficit are delayed in the SOD-1 PS line.

<p>(A) The SOD-1 (G93A)^PS mice (n = 8) have a delayed onset of weight decrease when compared to the original line (n = 17) (p<0.05). (B) Normal SOD-1 Tg mice start with a decrease in muscle strength at the age of 70 days. SOD-1 (G93A)^PS mice have normal muscular strength until the age of 310 days (p<0.05).</p

The SOD-1 (G93A) transgene in the SOD-1 (G93A)^PS line.

<p>(A) Genotyping of the SOD-1 (G93A)^PS mouse reveals a characteristic band from the SOD-1 (G93A) transgene. (B) Copy number of the SOD-1^G93A transgene is reduced in the SOD-1 (G93A)^PS mice by a factor of 6 when compared to normal SOD-1 (G93A) mice, littermates from the first SOD-1 (G93A)^PS mouse or from newly rederived SOD-1 (G93A) mice (p<0.05).</p

Listed are survival times (mean±SEM) in the SOD-1 (G93A)^PS line through 4 subsequent generations.

<p>Survival times are highly similar suggesting that the phenotype of the new line is stable.</p

SCD1 expression in ALS patient muscle and after nerve injury.

<p>(A) Expression of SCD1 and SCD5 in deltoid muscle biopsies from ALS patients and healthy subjects (CT, white columns), as identified by microarray analysis of the database deposited at <a href="http://www.ebi.ac.uk/arrayexpress/(accession" target="_blank">http://www.ebi.ac.uk/arrayexpress/(accession</a> number E-MEXP-3260) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0064525#pone.0064525-Pradat1" target="_blank">[12]</a>. ALS samples were obtained from muscle not clinically or electromyography affected (Unaff, orange columns) and from muscle with advanced pathology, characterized by reduced strength and neurogenic electromyography pattern (Aff, brown columns). *<i>P</i><0.05 (1-way ANOVA followed by Tukey's multiple comparison test, n = 4–10). (B) Expression of SCD1 in gastrocnemius following sciatic nerve axotomy (Axo) or crush at indicated post-operation days. Contralateral muscle expression is represented by 100% baseline. **<i>P</i><0.01, ***<i>P</i><0.001 (One sample t-test, n = 4–10).</p

Gene expression specific to the motor end plate in SCD1 knockout mice.

<p>Expression of AChR-α, AChR-γ, AChR-ε and MuSK in gastrocnemius and tibialis anterior from SCD1 knockout mice (brown columns) and wild-type littermates (white columns). *<i>P</i><0.05, ***<i>P</i><0.001 (Unpaired t-test, n = 4–11).</p

Metabolic phenotype of muscle from SCD1 knockout mice.

<p>(A) Expression of PGC1-α, PPARα and PDK4 in gastrocnemius and tibialis anterior from SCD1 knockout mice (brown columns) and wild-type littermates (white columns). *<i>P</i><0.05, **<i>P</i><0.01 (Unpaired t-test, n = 3–11). (B) Number of muscle fibers in tibialis anterior from SCD1 knockout mice (KO, brown column) and wild-type littermates (WT, white column). *<i>P</i><0.05 (Unpaired t-test, n = 7–10). (C) Distribution of the calibers of muscle fibers in tibialis anterior from SCD1 knockout mice (327 fibers, black circles) and wild-type littermates (283 fibers, white circles). Representative microphotographs of wild-type and knockout tibialis anterior are shown. (D) Averaged cross-sectional area of SDH-positive and SDH-negative fibers in tibialis anterior from SCD1 knockout mice (brown columns) and wild-type littermates (white columns). *<i>P</i><0.05, **<i>P</i><0.01 (Unpaired t-test, n = 7–10). (E) Number of SDH-positive (orange bars) and SDH-negative fibers (white bars) in tibialis anterior from SCD1 knockout mice (KO) and wild-type littermates (WT). ***<i>P</i><0.001 (Chi-square test, n = 283–327).</p

Muscle function recovery in SCD-deficient mice submitted to nerve crush.

<p>Restoration of muscle grip strength in SCD1 knockout mice (A) or MF-438 treated mice (C) (black circles) and corresponding control littermates (white circles) at the indicated post-operation times. ***<i>P</i><0.001 (2-way ANOVA, n = 4–12). Percentage of electromyography episodes of spontaneous activity in SCD1 knockout mice (Inset A) or MF-438 treated mice (Inset C) (KO or MF, brown columns) and corresponding control littermates (WT or CT, white columns) two weeks after sciatic nerve crush. *<i>P</i><0.05 (Unpaired t-test, n = 4–7). (B) Relative density of muscle fiber types in ipsilateral and contralateral tibialis anterior from SCD1 knockout mice and wild-type littermates two weeks after sciatic nerve crush. According to SDH histochemistry, fibers were classified as dark brown colored fibers with high metabolic oxidative capacity (brown columns), pale brown colored fibers with medium oxidative capacity (orange columns) and non-satined fibers (white columns). *<i>P</i><0.05 and ***<i>P</i><0.001 (1-way ANOVA followed by Tukey's multiple comparison test, (n = 4–6). (D) Kaplan-Meier curves showing the percentage of MF-438 treated mice (black circles) and control littermates (white circles) that started to exhibit a grip strength distinct from zero after initial total paralysis. Logrank test (n = 10–11). Inset D, averaged time at start of recovery in MF-438 treated mice (MF, brown column) and control littermates (CT, white column). *<i>P</i><0.05 (Unpaired t-test, n = 10–11).</p

Effects of MF-438 on metabolism and muscle function.

<p>(A) C16:1/C16:0 and C18:1/C18:0 fatty acid ratio in plasma from MF-438 treated mice (brown columns) and control littermates (CT, white columns). ***<i>P</i><0.001 (Unpaired t-test, n = 4–6). (B) Time course of respiratory quotient (RQ) before and after treatment with MF-438 at a dose of 10 mg/kg body mass/day (indicated by the black bar) (n = 4). Time course of body mass (C), muscle grip strength expressed as a percentage of day 0 (D), and specific grip strength, as determined by normalizing peak force to body mass (E), in mice fed regular chow (white circles) and mice fed regular chow supplemented with MF-438 (black circles). **<i>P</i><0.01 (2-way ANOVA followed by Bonferroni test, n = 5–6). (F) Expression of PGC1-α, AChR-α, and MuSK in gastrocnemius from MF-438 treated mice (brown columns) and control littermates (white columns). *<i>P</i><0.05 (Unpaired t-test, n = 3–9).</p