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. 2011 Sep;7(9):985-92.
doi: 10.4161/auto.7.9.16012. Epub 2011 Sep 1.

In vivo optical imaging of motor neuron autophagy in a mouse model of amyotrophic lateral sclerosis

Fengfeng Tian  1 , Nobutoshi Morimoto, WenTao Liu, Yasuyuki Ohta, Kentaro Deguchi, Kazunori Miyazaki, Koji Abe
Affiliations

In vivo optical imaging of motor neuron autophagy in a mouse model of amyotrophic lateral sclerosis

Fengfeng Tian et al. Autophagy. 2011 Sep.

Abstract

Autophagy is involved in the pathological process of motor neuron death in amyotrophic lateral sclerosis (ALS). We have generated a novel double transgenic (DTg) mouse line by mating a green fluorescent protein (GFP)-fused microtubule-associated protein 1 light chain 3 (LC3) transgenic (LC3-Tg) mouse and a G93A mutant human Cu/Zn superoxide dismutase (mSOD1) transgenic (mSOD1-Tg) mouse. In vivo imaging of autophagy with these novel DTg mice was conducted at 10 (presymptomatic), 17 (early symptomatic) and 19 (late symptomatic) weeks of age. Fluorescence imaging analysis revealed a strong fluorescent signal in vivo over the T₃-S₁ level at 17 and 19 weeks of age only in the DTg mice. Ex vivo autophagy imaging of spinal cord sections (20 μm) also showed a progressive increase of the fluorescence signal from 17 to 19 weeks in DTg mice in the anterior horn at the L₄-₅ level, and the fluorescence signals were clearly observed in the gray matter of the spinal cord with a progressive increase of the signal and decreases in large motor neurons. Protein gel blot analysis revealed maximum LC3-I and LC3-II expressions at 19 weeks, consistent with the results from the in vivo autophagy imaging experiment. This method could also be applied as a unique tool for clarifying the role of autophagy, and to monitor the pathologic processes involving autophagy not only in ALS, but also other neurological diseases.

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