Adipose-derived stem cell conditioned medium impacts asymptomatic peripheral neuromuscular denervation in the mutant superoxide dismutase (G93A) transgenic mouse model of amyotrophic lateral sclerosis
Affiliations: [a]
Department of Biomedical and Applied Sciences, Indiana University School of Dentistry, Indianapolis, IN, USA
| [b]
Department of Anatomy and Cell Biology, Indiana University School of Medicine, Indianapolis, IN, USA
| [c]
Department of Neurology, Indiana University School of Medicine, Indianapolis, IN, USA
| [d]
Department of Medicine, Indiana University School of Medicine, Indianapolis, IN, USA
| [e]
Roudebush VA Medical Center, Indianapolis, IN, USA
| [f] Division of Cardiovascular Medicine, Center for Regenerative Medicine, University of Florida, Gainesville, FL, USA
Correspondence:
[*]
Corresponding author: Chandler L. Walker, Ph.D., Assistant Professor, Department of Biomedical and Applied Sciences, Indiana University School of Dentistry, DS260A, 1121 W. Michigan Street, Indianapolis, IN 46202, USA. Tel.: +1 317 278 1402; E-mail: chalwalk@iu.edu.
Abstract: Background:Amyotrophic lateral sclerosis (ALS) is devastating, leading to paralysis and death. Disease onset begins pre-symptomatically through spinal motor neuron (MN) axon die-back from musculature at ∼47 days of age in the mutant superoxide dismutase 1 (mSOD1G93A) transgenic ALS mouse model. This period may be optimal to assess potential therapies. We previously demonstrated that post-symptomatic adipose-derived stem cell conditioned medium (ASC-CM) treatment is neuroprotective in mSOD1G93A mice. We hypothesized that early disease onset treatment could ameliorate neuromuscular junction (NMJ) disruption. Objective:To determine whether pre-symptom administration of ASC-CM prevents early NMJ disconnection. Methods:We confirmed the NMJ denervation time course in mSOD1G93A mice using co-labeling of neurofilament and post-synaptic acetylcholine receptors (AchR) by α-bungarotoxin. We determined whether ASC-CM ameliorates early NMJ loss in mSOD1G93A mice by systemically administering 200μl ASC-CM or vehicle medium daily from post-natal days 35 to 47 and quantifying intact NMJs through co-labeling of neurofilament and synaptophysin with α-bungarotoxin in gastrocnemius muscle. Results:Intact NMJs were significantly decreased in 47 day old mSOD1G93A mice (p < 0.05), and daily systemic ASC-CM prevented disease-induced NMJ denervation compared to vehicle treated mice (p < 0.05). Conclusions:Our results lay the foundation for testing the long-term neurological benefits of systemic ASC-CM therapy in the mSOD1G93A mouse model of ALS.
