Fig. 1.

Findings it the first cases point out a potential dystrophinopathy. a Pictures of the two first affected brothers (LRMD1 and 2), at 4 months of age, at time of their presentation to the neurology consultation of the Alfort school of veterinary medicine. Note the markedly plantigrade and palmigrade posture and the pelvic verticalisation. b Electromyographic recordings showing complex repetitive discharges observed in both animals, especially in the proximal appendicular muscles. c H&E staining of a biceps femoris biopsy taken from LRMD2 at 4 months of age showing a dystrophic profile: necrosis, regeneration, endomysial fibrosis, hypercontracted fibres, inflammatory foci and calcifications. d Dystrophin (rod-domain) immunostaining in LRMD1 and 2 biceps femoris compared to an unaffected dog. The images clearly show absence of signal delineating the membrane of the muscle fibres in LRMD dogs and presence of this signal in the images corresponding to the control dog. e Multiplex Western-blot confirming the absence of dystrophin immunoreactivity in both affected dogs using two different anti-dystrophin antibodies (Dys2, anti-C-terminal part, and Dys1, anti-rod domain). γ-Sarcoglycan was present in both affected dogs but levels were lower than in the healthy dog, probably because of disruption of the dystrophin-associated protein complex. The levels of other studied proteins were unmodified except for the α-sarcoglycan, for which the human antibody showed no cross-reactivity in canine muscles