G.P.19 Collagen VI genes in zebrafish skeletal muscle: Implications for collagen VI-myopathies

Ramanoudjame, Laetitita; Rocancourt, Claire; Lainé, Jeanne; Lyphout, Laura; Gartioux, C.; Schwartz, Marie-Elise; Cousin, Xavier; Allamand, V.

doi:10.1016/j.nmd.2012.06.089

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“…The col6a1 ama605003 stable line we generated, allowed us to study at the functional and histological level collagen VI-deficient fish up to 9 mpf for the first time. The col6a1 ama605003 zebrafish mutants exhibited histological and ultrastructural muscle alterations identical to the ones observed in muscle cell cultures [ 64 – 67 ] and biopsies [ 67 , 68 ] from patients with collagen VI deficiency, in mice [ 30 – 35 ], as well as in the transient zebrafish model generated by morpholino-based knock-down [ 36 , 37 ].…”

Section: Discussionmentioning

confidence: 96%

“…In mouse lines, each of the three genes encoding collagen VI has been individually inactivated and the corresponding alterations in muscle were extensively studied in these mutants [ 30 – 35 ]. Similarly, zebrafish models with different forms of altered collagen VI chains were obtained by the injection of morpholinos [ 36 , 37 ]. However, the morphants only allowed the study of transient phenotypes in embryos up to 2 days post-fertilization (dpf).…”

Section: Introductionmentioning

confidence: 99%

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A TALEN-Exon Skipping Design for a Bethlem Myopathy Model in Zebrafish

et al. 2015

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Presently, human collagen VI-related diseases such as Ullrich congenital muscular dystrophy (UCMD) and Bethlem myopathy (BM) remain incurable, emphasizing the need to unravel their etiology and improve their treatments. In UCMD, symptom onset occurs early, and both diseases aggravate with ageing. In zebrafish fry, morpholinos reproduced early UCMD and BM symptoms but did not allow to study the late phenotype. Here, we produced the first zebrafish line with the human mutation frequently found in collagen VI-related disorders such as UCMD and BM. We used a transcription activator-like effector nuclease (TALEN) to design the col6a1ama605003-line with a mutation within an essential splice donor site, in intron 14 of the col6a1 gene, which provoke an in-frame skipping of exon 14 in the processed mRNA. This mutation at a splice donor site is the first example of a template-independent modification of splicing induced in zebrafish using a targetable nuclease. This technique is readily expandable to other organisms and can be instrumental in other disease studies. Histological and ultrastructural analyzes of homozygous and heterozygous mutant fry and 3 months post-fertilization (mpf) fish revealed co-dominantly inherited abnormal myofibers with disorganized myofibrils, enlarged sarcoplasmic reticulum, altered mitochondria and misaligned sarcomeres. Locomotion analyzes showed hypoxia-response behavior in 9 mpf col6a1 mutant unseen in 3 mpf fish. These symptoms worsened with ageing as described in patients with collagen VI deficiency. Thus, the col6a1ama605003-line is the first adult zebrafish model of collagen VI-related diseases; it will be instrumental both for basic research and drug discovery assays focusing on this type of disorders.

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