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. Author manuscript; available in PMC: 2015 Aug 1.
Published in final edited form as: Neurobiol Dis. 2014 Apr 28;68:78–90. doi: 10.1016/j.nbd.2014.04.013

Figure 5. Inhibition of mitochondrial transport interferes with the ability of cytNmNAT1 to protect axons against hydrogen peroxide or OGD.

Figure 5

(A) Neurons were transfected with either soluble GFP (control) or dominant-negative Miro (dnMiro) construct and their axonal integrity was examined 20 h later. Transiently expressing dnMiro did not affect axonal integrity (8 and 10% severely damaged respectively P>0.05). (B) Neurons were electroporated with cytNmNAT1-GFP before plating. After 7 days in culture they were transfected with either dnMiro-RFP or soluble RFP alone, were then challenged with either 100 μM hydrogen peroxide for 1 h (C) or OGD for 4 h (D). The axons of neurons expressing cytNmNAT1 and a soluble fill showed minimal signs of damage following either insult (C & D left column). In marked contrast, the axons of neurons expressing dominant-negative Miro showed extensive beading and fragmentation (C & D middle column). When neurons were exposed to hydrogen peroxide, 61% of the axons were severely degenerated in neurons expressing dominant-negative Miro and cytNmNAT1, compared with only 14% in neurons expressing cytNmNAT1 alone. Similarly, in neurons expressing both dnMiro and cytNmNAT1, 63% of axons were severely degenerated after OGD, compared with only 2.5% in neurons with cytNmNAT1 alone (C & D right column). Axonal damage was assessed 6 h after hydrogen peroxide and 24 h after OGD. *** p< 0.001. Scale bar=30 μm.