Cytoskeletal structure in cells harboring two mutations: R133C in NOTCH3 and 5650G>A in mitochondrial DNA

We have previously described a patient with cerebral autosomal-dominant arteriopathy with subcortical infarcts and leucoencephalopathy (CADASIL) caused by R133C mutation in NOTCH3 and with a concomitant myopathy caused by a G to A point mutation at base pair 5650 (5650G>A) in the gene encoding tRNAAla in mitochondrial DNA (mtDNA). In the present study, we have examined the morphology of the cytoskeletal components in fibroblasts and myoblasts of this patient. Immunolabeling revealed that tubulin network was sparse and formed asters in these cells, whereas no changes were found in actin and vimentin networks in comparison to the control cell lines. Furthermore, mitochondria were less abundant and the branches of the mitochondrial network were reduced in number. Muscle histochemical analysis showed ragged red fibres (RRFs) and cytochrome c oxidase (COX)-negative fibres. The mean proportion of mtDNA with 5650G>A was lower in histologically normal muscle fibres than in the COX-negative fibres and in the RRFs. These findings suggest that 5650G>A is a pathogenic mtDNA mutation. However, the changes in tubulin network and mitochondrial distribution in patient fibroblasts and myoblasts cannot solely be explained by this mutation.