Fig. 8

Light microscopy findings of extensor digitorum longus muscle biopsy in the right leg of Patient 4. Variation in fibre calibre, nuclear internalization (white arrow in a), fibre splitting (yellow arrows in a, and b), multiple splitting in degenerating fibre (white arrow in b), rimmed vacuole (yellow arrow in c), small dark type 1 fibres (yellow * in d), core-like oxidative irregular reactions (yellow arrows in e, f, and g), and intrasarcoplasmic acid phosphatase activity (yellow arrow in h). Immunohistochemistry with complete negative intrassarcoplasmic telethonin reaction (arrow in i, compared to the normal control (inset arrow in i). Serial sections of a non-necrotic muscle fibre (same fibre marked with yellow * in j, k, and l) with focal secondary reduction in membrane dysferlin expression (yellow arrow in k) in the sarcolemmal membrane, compared to strong sarcolemmal membrane reaction of the adjacent fibres, and compared to normal dystrophin reaction in serial sections of the same fibre (yellow arrow in l) as proof of membrane integrity. (a HE 200x, b HE 400x. c HE 400x, d ATPase pH4.6 100x, e SDH 200x, f COX-SDH 200x, g NADH 400x, h Acid phosphatase 200x, Immunoperoxidase Antibody anti-telethonin (G-11 sc-25327, Santa Cruz Biotechnology, Inc. at 1:50 dilution) 200x, Patient (i), and 400x Control (inset in i), j. HE 400x, k. Immunoperoxidase Antibody anti-dysferlin (HAM1/7B6, Vector Laboratories) 400x, and l. Immunoperoxidase Antibody anti-dystrophin carboxy-terminal (NCL-DYS2, Novocastra, Leica) 400x)