Early Functional Deficit and Microglial Disturbances in a Mouse Model of Amyotrophic Lateral Sclerosis methods
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Methods and Findings
We have evaluated locomotor function of hSOD1 G93A mice through dynamic walking patterns and spontaneous motor activity analysis. We detected early functional deficits that redefine symptoms onset at 60 days of age, i.e. 20 days earlier than previously described. Moreover, sequential combination of these approaches allows monitoring of motor activity up to disease end stage. To tentatively correlate early functional deficit with cellular alterations we have used flow cytometry and immunohistochemistry approaches to characterize neuromuscular junctions, astrocytes and microglia. We show that (1) decrease in neuromuscular junction's number correlates with motor impairment, (2) astrocytes number is not altered at pre- and early-symptomatic ages but intraspinal repartition is modified at symptoms onset, and (3) microglia modifications precede disease onset. At pre-symptomatic age, we show...
Locomotor function is altered at 60 days of age in hSOD1 G93A mice
Using an automated gait analysis system, we report early functional deficits that redefine symptoms onset at 60 days of age, i.e. 20 days earlier than previously described. Our results confirm and extend recent data obtained with the digigait treadmill video based system on the same transgenic strain. However, noteworthy is that the CatWalk™, through the dynamic analysis of walking patterns, and conversely to the treadmill, does not induce any physical exercise that could affect the lifespan of the mice,,. Moreover, the reproducibility of the digigait analysis system is still under debate,. The CatWalk™ system seems thus better adapted for the evaluation of therapeutic approaches. In a recent study, early motor symptoms had been detected at 45 days of age using a rotarod in SOD1 G93A mice on a C57/BL/6 background. In the same study modifications of step patterns were...
Methods
Experimental procedures followed the European legislative, administrative and statutory measures for animal experimentation (86/609/EEC) and the Declaration of Helsinki. The study was approved by the "Direction des Services Vétérinaires de l'Hérault", France (authorization number 34118) and ratified by the "Préfecture de l'Hérault", France. Every effort was made to minimize the number of animals and their suffering. Transgenic mice carrying the G93A human SOD1 mutation B6SJL-Tg (SOD1-G93A) 1Gur/J (ALS mice) were purchased from The Jackson Laboratory (Bar Harbor, ME, USA) and bred on a B6SJL background. Transgenic mice were identified by PCR and housed in controlled conditions (hygrometry, temperature and 12 h light/dark cycle). Only males were used and litter-matching between groups were done as much as possible. Record of the weight was done once...
Measurement outputs
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Gait recovery and locomotor function
- Raw artifact
- Per-run gait capture with paw placement, timing, and stride features for each animal
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- Cleaned gait metrics table and recovery trend summary across timepoints
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- Group comparisons of gait indices, stride metrics, or recovery curves
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Quantify the primary readouts for this experiment: Gait recovery and locomotor function.
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inferred from protocolReporting output
Report representative outputs alongside summary comparisons for Gait recovery and locomotor function.
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Evidence quotes (3)
We have evaluated locomotor function of hSOD1 G93A mice through dynamic walking patterns and spontaneous motor activity analysis. We detected early functional deficits that redefine symptoms onset at 60 days of age, i.e. 20 days earlier than previously described. Moreover, sequential combination of these approaches allows monitoring of motor activity up to disease end stage. To tentatively correlate early functional deficit with cellular alterations we have used flow cytometry and immunohistochemistry approaches to characterize neuromuscular junctions, astrocytes and microglia. We show that (1) decrease in neuromuscular junction's number correlates with motor impairment, (2) astrocytes number is not altered at pre- and early-symptomatic ages but intraspinal repartition is modified at symptoms onset, and (3) microglia modifications precede disease onset. At pre-symptomatic age, we show a decrease in microglia number whereas at onset of the disease two distinct microglia sub-populations emerge.
Using an automated gait analysis system, we report early functional deficits that redefine symptoms onset at 60 days of age, i.e. 20 days earlier than previously described. Our results confirm and extend recent data obtained with the digigait treadmill video based system on the same transgenic strain. However, noteworthy is that the CatWalk™, through the dynamic analysis of walking patterns, and conversely to the treadmill, does not induce any physical exercise that could affect the lifespan of the mice,,. Moreover, the reproducibility of the digigait analysis system is still under debate,. The CatWalk™ system seems thus better adapted for the evaluation of therapeutic approaches. In a recent study, early motor symptoms had been detected at 45 days of age using a rotarod in SOD1 G93A mice on a C57/BL/6 background. In the same study modifications of step patterns were identified around P75 using CatWalk™ system. One of the drawbacks of the use of the rotarod in the assessment of a therapeutic approach is its influence on the pathophysiology of the disease, such as increases of the lifespan and cell proliferation that have been reported in a mouse model of...
Experimental procedures followed the European legislative, administrative and statutory measures for animal experimentation (86/609/EEC) and the Declaration of Helsinki. The study was approved by the "Direction des Services Vétérinaires de l'Hérault", France (authorization number 34118) and ratified by the "Préfecture de l'Hérault", France. Every effort was made to minimize the number of animals and their suffering. Transgenic mice carrying the G93A human SOD1 mutation B6SJL-Tg (SOD1-G93A) 1Gur/J (ALS mice) were purchased from The Jackson Laboratory (Bar Harbor, ME, USA) and bred on a B6SJL background. Transgenic mice were identified by PCR and housed in controlled conditions (hygrometry, temperature and 12 h light/dark cycle). Only males were used and litter-matching between groups were done as much as possible. Record of the weight was done once a week from 56 days onward.
Machine-readable layer
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