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Satoshi Yokota, Yuji Takihara, Shogo Arimura, Seiji Miyake, Yoshihiro Takamura, Nagahisa Yoshimura, Masaru Inatani; Live imaging for mitochondrial axonal transport in the retinal ganglion cells after laser-induced axonal damage. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):2441.
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© ARVO (1962-2015); The Authors (2016-present)
Axonal damage is a critical event for induces the apoptosis of retinal ganglion cells (RGCs) in glaucoma optic neuropathy. We evaluated whether visualization for mitochondrial transport contributes to the prediction for the RGC apoptosis after axonal damage.
Rat RGCs on postnatal day 3 were purified by a two-step immunopanning method. Mitochondria in cultivated RGCs were stained with Rhodamine 123. The dynamics of mitochondria in the axons was quantified on cultivating day 6 by time-lapse imaging. Axonal damage was created by laser axon injury using laser microdissection system. The numbers and velocity of moving mitochondria were compared among 5 minute, a half day and 3 days after laser-induced axonal damage. RGC death after laser-induced axonal damage was determined by labeling with ethidium homodimer-1.
The number of moving mitochondria and the mean velocity of mitochondrial movement in the RGC axons were 1.83 + 0.76 /axon · min and 0.44 + 0.23 µm/sec. After laser-induced axonal damage, the numbers were 0.49 + 0.59 and 0.55 + 0.67 /axon · min on 5 min and a half day after laser induced damage, respectively. The velocities were 0.34 + 0.21 and 0.42 + 0.26 µm/sec on 5 min and a half day after laser damage, respectively. Compared with mitochondrial movement without laser-induced axonal damage, the numbers and velocities were significantly decreased 5 min after laser-induced axonal damage (P < 0.0001 and P = 0.0001, respectively). 35.3% of RGCs with laser-induced axonal damage showed transient decrease of the velocity (the recovered group). The number of ethidium homodimer-1-positive RGCs was significantly lower in the recovered group (16.7%) than the non-recovered group (72.7%) (P=0.006).
Mitochondrial transport in RGCs is disturbed after laser-induced axonal damage. Irreversible deterioration of mitochondrial transport shows correlation with RGC apoptosis.
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