Figure 2
Local pharmacological inhibition of mitochondrial fission prevented axonal damage.
(a–l) Paired morphology panel of DIV10 CGN in microfluidic chambers. The somatic chamber was probed with Hoechst nuclear staining (a,c,e,g,i,k) and axonal endings in the axonal chamber immuno-stained with beta3-tubulin (b,d,f,h,j,l). (a,b), 10 DIV CGN grown in high glucose (HG) conditions. (c,d) Application of axonal rotenone (Rot HG) in the axonal chamber did not trigger axonal degeneration in HG condition after 24 hours of treatment. (e,f) Low glucose (LG) condition was innocuous. (g,h) Application of axonal rotenone in LG triggered axonal degeneration. (i,j) Application of 10 μM Mdivi-1 on axons inhibited the action of rotenone, but 10 μM SP600125 did not (k,l). (Scale bar: 10 μm). (m) Quantification of axonal degeneration. Each experiment was conducted at 3 times independently in triplicates and data were analyzed using ANOVA statistical method. (n) Representative images of both proximal (somatic chambers) and distal (axonal chamber) axonal mitochondrial morphology upon TOM20 immunostaining after rotenone insult and pharmacological blockade.
