DCA Effects in ALS Models Figure 2. DCA recovers mitochondrial respiration rate and controls proliferation in SOD1G93A astrocytes. A) Representative immunoblot for PDH-E1a(pSer293), total PDH-E1a, and b-actin of lysates from non Tg and SOD1G93A astrocytes after 24 h treatment with DCA or vehicle as described in Methods. B) Quantification of the PDH-E1a(pSer293) to total PDH-E1a ratio between relative densitometric levels normalized against vehicle-treated non Tg astrocytes. C) Calculated respiratory control ratio (RCR) for mitochondria from non Tg or SOD1G93A-bearing astrocytes treated with DCA or vehicle as indicated. D) Percentage of BrdU immunoreactive nuclei of non Tg and SOD1G93A astrocytes after 24 h treatment with DCA. Data for panels B, C, and D are expressed as mean 6 SEM from three independent experiments performed in duplicate. *p,0.05, significantly different from non Tg control. **p,0.05, significantly different from SOD1G93A control. doi:10.1371/journal.pone.0034776.g002 cantly improved grip strength performance at the end stage of the disease (from 100 days of age onward) in male mice compared to control transgenic ones (Fig. 5A). Grip strength did not improve in females (data not shown). Because there is a correlation between the number of active motor units and the force produced by a muscle, we decided to observe the neuromuscular junction (NMJ) Figure 3. DCA prevents SOD1G93A astrocyte neurotoxicity to motor neurons. Motor neuron survival 72 h after plating either on non Tg or SOD1G93A-bearing astrocytes pretreated with DCA or vehicle as indicated. Data are expressed as percentage of non Tg control, mean 6 SEM from four independent experiments. *p,0.05, significantly different from non Tg control. **p,0.05, significantly different from SOD1G93A control. doi:10.1371/journal.pone.0034776.g003 morphology of two muscles with different fiber composition, extensor digitorum longus (EDL) and soleus. The NMJs of EDL muscle in DCA-treated mice displayed normal size (measured as total area) and shape, which is in agreement with the improvement of grip strength (Fig. 5B). In control SOD1G93A mice, the receptor area decreased in size, as did the spaces between synaptic regions, leading to an overall compaction of the junction. A 20-day DCA treatment from 70 days of age onward showed a significant increase in junction area in EDL muscles compared with untreated mice. Neuromuscular junctions in soleus muscles did not show significant improvements with DCA treatment (data not shown). DCA treatment improved mitochondrial function in the spinal cord of SOD1G93A mice To assess whether DCA improved mitochondrial function in vivo, high-resolution respirometry was monitored in mechanically dissociated spinal cords obtained from mice after 20 days of DCA administration (from 70 days of age). The RCR value calculated for untreated SOD1G93A mice spinal cords was significantly reduced compared to non Tg littermates according to previous reports [20]. In contrast, DCA-treated SOD1G93A mice showed a significant increase in the RCR compared to the untreated group (Fig. 6). DCA reduced motor neuron loss and astrocyte reactivity in the spinal cord of SOD1G93A mice We sought to determine the effect of DCA administration in the loss of motor neurons in the spinal cord. We noted the already described reduced motor neuron somas at lumbar spinal segments PLoS ONE | www.plosone.org 3 April 2012 | Volume 7 | Issue 4 | e34776