In neurons, activation of GluRs induces COX 2 expres sion which can contribute to excitotoxic neuronal death. As a way to establish no matter whether a comparable result of GluR activation occurs for oligodendrocytes, dispersed cultures have been taken care of with sub selelck kinase inhibitor lethal doses of KA and also the quantity of COX two expression examined by immunofluo rescent confocal microscopy. As noticed in Figure 5, cultures taken care of with KA present a robust induction of COX 2 24 hrs after KA treatment when in comparison with handle cul tures. This is often constant which has a prospective purpose of COX 2 in excitotoxic death of oligodendrocytes. COX two inhibitors shield towards excitotoxic death of oligodendrocytes in dispersed cultures The possible protective result in the COX two inhibitor CAY 10404 was examined in dispersed oligodendrocytes handled with KA. As witnessed in Figure six, treatment method with COX 2 inhibitor resulted inside a one.
five fold maximize in surviv ing KA taken care of oligodendrocytes at 24 hours. This outcome signifies that COX two expression in oligodendrocytes increases excitotoxic death. Improved expression of COX two in oligodendrocytes enhances excitotoxic death The previous selleckchem outcomes with COX 2 inhibitors present sup portive proof to get a part for COX two in excitotoxic death of oligodendrocytes. Nonetheless, 1 prospective caveat to these success is the fact that COX 2 inhibitors may well have off target routines that could market protective results inde pendent of COX 2 inhibition. Therefore, we implemented genetic manipulation to alter COX two expression as a way to assess irrespective of whether alterations inside the expression have an result on oli godendrocyte vulnerability to excitotoxic death. A trans genic mouse was created that was created to raise expression of COX 2 particularly in oligodendrocytes. This was attained by linking the human COX 2 gene downstream in the oligodendrocyte promoter for that CNPase gene.
The human COX 2 gene has in essence exactly the same catalytic properties because the endoge nous mouse COX two gene, but is made up of some distinct amino acid sequences

that make it uniquely detectable with human COX 2 unique antibodies. When oligodendrocytes have been isolated from these trans genic mice and probed with an antibody for COX 2, it had been appar ent that the oligodendrocytes derived from the transgenic mice exhibit a robust increase in COX 2 expression com pared to wild variety oligodendrocytes. So as to test our hypothesis that COX 2 expression in oligoden drocytes increases sensitivity to excitotoxic death, these COX 2 transgenic oligodendrocytes have been in comparison to wild style oligodendrocytes for their susceptibilities to KA induced excitotoxic death. As viewed in Figure eight, the KA concentration response curve for your transgenic COX two oligodendrocytes was shifted towards the left when when compared to that viewed with wild style oligodendrocytes, indicating the transgenic COX two oligodendrocytes are additional sensitive to KA induced excitotoxic death. Comparison with the concentrations of KA needed to destroy 50% on the cells signifies the COX two transgenic oli godendrocytes are eight fold extra sensitive to KA com pared to wild form.