Characterization of dystrophin deficient rats: a new model for Duchenne muscular dystrophy.

Authors:
Larcher T, Lafoux A, Tesson L, Remy S, Thepenier V, François V, Le Guiner C, Goubin H, Dutilleul M, Guigand L, Toumaniantz G, De Cian A, Boix C, Renaud JB, Cherel Y, Giovannangeli C, Concordet JP, Anegon I, Huchet C.
In:
Source: PLoS ONE
Publication Date: (2014)
Issue: 9(10): e11037
Research Area:
Basic Research
Molecular Biology
Cells used in publication:
C6
Species: rat
Tissue Origin: brain
Platform:
4D-Nucleofector® X-Unit
Experiment

Each subunit of TALE nuclease (0.75 and 1.5 µg each) was nucleofected into 4.105 C6 cells (Sigma) in 20 µL of V solution using 4D-Nucleofector FF-137 program. Cells were grown into 12 well dishes and collected 48 h after nucleofection. Lonza summary: The authors generated a novel trustable rat model for the Duchenne muscular dystrophy genetic disease or DMD. They mutated the dystrophin gene using TALENs, first validated into rat C6 cells using the 4D Nucleofector technology then into rat zygotes using microinjection technique. This paper shows a good example of how relevant and cheaper animal disease models could be generated using Nucleofection in combination with genome editing tools.

Abstract

A few animal models of Duchenne muscular dystrophy (DMD) are available, large ones such as pigs or dogs being expensive and difficult to handle. Mdx (X-linked muscular dystrophy) mice only partially mimic the human disease, with limited chronic muscular lesions and muscle weakness. Their small size also imposes limitations on analyses. A rat model could represent a useful alternative since rats are small animals but 10 times bigger than mice and could better reflect the lesions and functional abnormalities observed in DMD patients. Two lines of Dmd mutated-rats (Dmdmdx) were generated using TALENs targeting exon 23. Muscles of animals of both lines showed undetectable levels of dystrophin by western blot and less than 5% of dystrophin positive fibers by immunohistochemistry. At 3 months, limb and diaphragm muscles from Dmdmdx rats displayed severe necrosis and regeneration. At 7 months, these muscles also showed severe fibrosis and some adipose tissue infiltration. Dmdmdx rats showed significant reduction in muscle strength and a decrease in spontaneous motor activity. Furthermore, heart morphology was indicative of dilated cardiomyopathy associated histologically with necrotic and fibrotic changes. Echocardiography showed significant concentric remodeling and alteration of diastolic function. In conclusion, Dmdmdx rats represent a new faithful small animal model of DMD.