AI Summary & Key Takeaways
AI InsightThus for the first time a functional link between Als2, and NF-κB pathway over UXT has been confirmed in primary spinal motor neuron culture , and the role played by key mo-lecules as IL8, and A20 on NF-kB pathways has been demonstrated.
📌 3 Key Takeaways
Özet
Aim: Amyotrophic Lateral Sclerosis (ALS) is a neurodegenerative di-sorder affecting motor neurons. Als2 is one of the genes that cause ALS, and its mutations leads to loss of function in alsin protein. The interaction between alsin, and UXT protein has been documented in vitro. UXT is a cofactor in NF-κB pathway. Our aim is to investigate the potential effect of silencing of Als2 on the activity of NF-κB path-way in primary motor neurons. Method: Neurons were demonstrated immunocytochemically via reaction with anti-ChAT and anti-βIII tubulin primary antibodies in spinal motor neuron culture of adult BalbC mice. Als2 was silenced using RNAi method and confirmed by qRT-PCR. Expressions of UXT, A20 and IL8 in motor neurons where Als2 was silenced were deter-mined by qRT-PCR. TNF-α, an NF-κB activator, was also applied and alterations in the expressions of A20 and IL8 were measured using qRT-PCR.Results: Expressions of Als2, UXT, and IL8 were reduced by 74.3%, 40%, and 84.4% by RNAi in immunocytochemically characterized spinal motor neurons (for all, p<0.0001). whereas A20 expression increased by 65% (p<0.0001). When TNF-α was applied to neurons, IL8 expression increased by 17-fold, but the increase in A20 was only 29.5%.Conclusion: In the light of these findings, correlation between exp-ressions of Als2 and UXT has been proven. Decreases in the levels of Als2 and UXT cause a reduction in IL8 expression, whereas expression of A20 increases. This indicates that silencing of Als2 might reduce inflammatory response while activating pro-apoptotic signals. When NF-κB pathway has been activated by TNF-α, a considerable increa-se in IL8 has been observed. Thus for the first time a functional link between Als2, and NF-κB pathway over UXT has been confirmed in primary spinal motor neuron culture , and the role played by key mo-lecules as IL8, and A20 on NF-kB pathways has been demonstrated.