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Surprisingly, ß-actin was not required for motor neuron viability or neuromuscular junction maintenance.
Motor neuron viability in Actb-MNsKO mice was assessed by quantifying Nissl stained motor neurons in the ventral horns of spinal cord sections.
Surprisingly, motor neuron specific ß-actin knock-out (Actb-MNsKO) mice exhibited no overt phenotype, with motor neuron viability, NMJ, and muscle performance all preserved in comparison to controls.
At 6 and 12 months of age, no significant differences were observed in the number of motor neurons per section in Actb-MNsKO compared to controls, indicating that ß-actin is not required for motor neuron viability in vivo (Figure 2A C).
These susceptibility factors may increase the vulnerability of motor neuron viability during disease.
The results thus far indicate that Fz3 inactivation could lead either to a primary defect in motor neuron viability that secondarily impairs axonal growth or a primary defect in axonal growth that secondarily impairs motor neuron viability.
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Having developed an in vitro screening cascade capable of identifying molecules that reduce oxidative stress and increase viability of motor neuron cells expressing an ALS-associated mutant of SOD1, we used the methodology to screen the Spectrum Collection of 2000 molecules.
The hits have been further refined in subsequent assays of oxidative stress and viability in motor neuron cells expressing an ALS-associated SOD1 mutation (G93A), and compounds likely to be able to access the CNS were selected based on their predicted biochemical properties.
Together these data suggest that both increased and decreased levels of TDP-43 initiate a loss-of-function phenotype, resulting in decreased neuronal viability and subsequent degenerative cell loss that characterize motor neuron diseases like ALS and FTLD.
Since animal LRP4 antibodies reduce the neuronal viability in cell culture, 18 LRP4 autoantibodies with access to motor neurons could potentially cause motor neuron degeneration.
Cell viability assays conducted over a period of 1 week showed that expression of Met-TDP-35 induced motor neuron death with a 50%% loss at day 5 post-microinjection, compared to full-length TDP-43, which exhibited no toxicity with a similar cell viability curve as neurons expressing the empty vector (Fig. 4b).
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