A new method of genotyping MDX^4CV mice by PCR‐RFLP analysis

Abstract

Introduction The mdx4cv mouse is a common model to study Duchenne muscular dystrophy (DMD). The most utilized methodology to identify the genotype of these mice is Sanger DNA sequencing. Methods Here, we provide a simple, cost-effective alternative approach to identify the wildtype, heterozygous, or homozygous/hemizygous genotypes of these mice, using commonly available laboratory equipment and reagents. Results Our technique exploits a restriction fragment length polymorphism (RFLP) that is generated by the point mutation found in exon 53 of mdx4cv mice. Discussion This technique can benefit laboratories that require complex breeding strategies involving mdx4cv mice.

Proteomic and cell biological profiling of the renal phenotype of the mdx-4cv mouse model of Duchenne muscular dystrophy

“…However, due to the extremely restricted amounts of patient tissue biopsy material available to study genetic diseases of childhood, animal models have been widely employed to study the molecular pathogenesis of X-linked muscular dystrophy (McGreevy et al, 2015;Rodrigues et al, 2016;Wilson et al, 2017). In analogy, we have used here kidney specimens from the established mdx-4cv mouse model of X-linked muscular dystrophy (Banks et al, 2010;Im et al, 1996;Tichy and Mourkioti, 2017). Genetic mdxtype mouse models have previously been used for studying cellular abnormalities in the kidney (Gusel'nikova et al, 2018) and the evaluation of kidney toxicity in relation to pharmacological applications (Reay et al, 2015) and experimental exon-skipping therapy (Zhang et al, 2015) to treat muscular dystrophy.…”

Mass Spectrometric Profiling of Extraocular Muscle and Proteomic Adaptations in the mdx-4cv Model of Duchenne Muscular Dystrophy

“…Genetic rearrangements in the DMD gene cause the almost complete loss of the full-length dystrophin isoform Dp427-M [ 21 ] and the simultaneous disintegration of the dystrophin-glycoprotein complex [ 22 ]. Established genetic animal models of dystrophinopathy, such as the mdx-4cv mouse [ 65 , 66 , 67 ], reflect many of the multifaceted and body-wide alterations seen in Duchenne patients, including necrosis, fibrosis and inflammation in the diaphragm muscle [ 39 , 68 ], cardiomyopathic changes [ 69 ] and neuronal deficiencies [ 70 ], as well as secondary abnormalities in the liver [ 71 ], kidney [ 48 ], and spleen [ 40 ]. The molecular and cellular pathogenesis of muscular dystrophy is also mirrored by characteristic protein changes in biofluids, such as mdx-4cv serum, urine and saliva [ 72 , 73 , 74 ].…”

Proteome-wide Changes in the mdx-4cv Spleen due to Pathophysiological Cross Talk with Dystrophin-Deficient Skeletal Muscle

“…Hence, in analogy to progressive forms of human dystrophinopathy, this animal model almost completely lacks the full-length dystrophin isoform Dp427-M ( Partridge, 2013 ). An alternative mdx -type mouse model ( mdx-4cv ) was generated by chemical mutagenesis with N-ethyl-N-nitrosourea ( Chapman et al., 1989 ) and resulted in a C to T transition at base 7,916 in exon 53, generating an ochre codon ( Banks et al., 2010 ; Shin et al., 2011 ; Tichy and Mourkioti, 2017 ). Importantly, the mdx-4cv model is typified by an approximately 10-fold lower rate of dystrophin-positive revertant fibers as compared with the conventional mdx-23 mouse ( Danko et al., 1992 ; Im et al., 1996 ; Judge et al., 2006 ).…”