Prolonged pegfilgrastim therapy increased the survival of SOD1 mice. When compared to their vehicle controlled littermates, pegfilgrastim increased the life expectancy from 173 6. 7 days to 185 6. 7 days with the range of 3 23 days. The time of onset did not than differ between the groups and the time from the onset to death was thus prolonged with Inhibitors,Modulators,Libraries the pegfil grastim treatment. The increased survival time was accompanied by improved performance in a wire hang behavioral test indicating more sustained motoric capacity in GCSF treated mutant SOD1 mice compared to vehicle treated littermates. GCSF conducts neuroprotection in vitro In Inhibitors,Modulators,Libraries order to uncover cellular and molecular mechanisms of GCSF mediated neuroprotection we tested the action of GCSF on primary neuronal culture.

Spinal cord neu ron culture consisted of NeuN positive neurons and to a lesser extent, SMI 32 Inhibitors,Modulators,Libraries positive motoneurons, and the lat ter defined by the large size of the cell body and the long axon typical to motoneurons in spinal cord cultures. On non stimulated neuronal cul tures, the treatment with GCSF increased the Akt phos phorylation as shown earlier in NSC34 secondary cells. When spinal Inhibitors,Modulators,Libraries cord neurons were exposed to glutamate induced excitotoxicity in vitro, GCSF reduced neuronal cell death. GCSF did not have any effect on neuron survival in control conditions. When spinal cord neuron culture was prepared from mutant SOD1 mice we discovered that the general cell viability was slightly reduced in mutant SOD1 neurons and GCSF could alleviate the compro mised cell viability.

However, unlike wt cells, GCSF did not protect mutant SOD1 neurons from glutamate neurotoxicity. Next, to determine the effect of long term pegfilgras tim treatment in vivo, spinal cord sections were analyzed by immunohistochemistry. The neurodegeneration was evident in mutant SOD1 mice when compared to wt mice, as evaluated by reduced Inhibitors,Modulators,Libraries immunoreactivity for neu ronal markers, Even though GCSF had cer tain neuroprotective properties in vitro, long term pegfilgrastim treatment did not significantly increase spinal cord neuron survival in mutant SOD1 mice in vivo. GCSF with sustained activity attenuates inflammation in vivo When the inflammation status was analyzed from the spinal cords of the same mice, long term pegfilgrastim treatment indeed decreased astrogliosis and microgliosis in the ventral horn of the spinal cord as determined with GFAP and Iba 1 immunostain ing, respectively. We further examined whether reduction of inflammation in the spinal cord could be detected in the level of inflammatory media tors. Firstly, quantitative PCR results selleck chem showed that the expression of TNFa was upregulated and that iNOS was downregulated in mutant SOD1 mice spinal cords compared to wt mice.