Visualization of Dystrophic Muscle Fibers in Mdx Mouse by Vital Staining with Evans Blue: Evidence of Apoptosis in Dystrophin-Deficient Muscle

Matsuda, Ryoichi; Nishikawa, Akio; Tanaka, Hiroaki

doi:10.1093/jb/118.5.959

Cited by 332 publications

(276 citation statements)

References 25 publications

(27 reference statements)

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“…18 In these experiments, animals were killed 6 h after Evans Blue injection. Liver sections from the animals were made and examined under a fluorescence microscope.…”

Section: Membrane Pores Reseal With Timementioning

confidence: 99%

Hydroporation as the mechanism of hydrodynamic delivery

et al. 2004

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We have reported that a rapid tail vein injection of a large volume of plasmid DNA solution into a mouse results in high level of transgene expression in the liver. Gene transfer efficiency of this hydrodynamics-based procedure is determined by the combined effect of a large volume and high injection speed. Here, we show that the hydrodynamic injection induces a transient irregularity of heart function, a sharp increase in venous pressure, an enlargement of liver fenestrae, and enhancement of membrane permeability of the hepatocytes. At the cellular level, our results suggest that hepatic delivery by the hydrodynamic injection is accomplished by the generation of membrane pores in the hepatocytes.

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“…18 In these experiments, animals were killed 6 h after Evans Blue injection. Liver sections from the animals were made and examined under a fluorescence microscope.…”

Section: Membrane Pores Reseal With Timementioning

confidence: 99%

Hydroporation as the mechanism of hydrodynamic delivery

et al. 2004

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“…The day before, mice received an intraperitoneal injection of Evans blue, a dye that penetrates degenerating fibers. 29 Frozen sections of quadriceps muscles were stained with hematoxylin and eosin (H&E) (Figure 2b and d). Dystrophin or a-sarcoglycan expression was determined by immunochemistry and muscle damage was evaluated by Evans blue uptake (Figure 2c …”

Section: Application Of Aav Gene Transfer Protocols To Rescue MDX Andmentioning

confidence: 99%

Noninvasive monitoring of therapeutic gene transfer in animal models of muscular dystrophies

et al. 2005

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Muscular dystrophies are a genetically and phenotypically heterogeneous group of degenerative muscle diseases. A subset of them are due to genetic deficiencies in proteins which form the dystrophin-associated complex at the membrane of the myofibers. In this report, we utilized recombinant adeno-associated virus containing a U7 cassette carrying an antisense sequence aimed at inducing exon skipping of the dystrophin gene or containing the a-sarcoglycan gene to alleviate the dystrophic phenotype of the mdx and Sgca-null mice, respectively. As these diseases are characterized by cycle of degeneration/regeneration, we postulated that a reporter gene coadministered at the time of the treatment would make it possible to follow the extent of muscle repair. We observed that the murine secreted alkaline phosphatase (muSeAP) level was very much lower in these animal models than in normal mice. Upon treatment of the dystrophic muscle by gene transfer, the level of muSeAP was restored and correlated with the expression of the therapeutic transgene and with the level of muscle improvement. The system described here provides a simple and noninvasive procedure for monitoring the outcome of a therapeutic strategy involving cell survival.

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“…As DMD patients live longer because of improved multidisciplinary patient care, rescuing dystrophin expression in cardiac muscle becomes more critical for their longevity and quality of life. [14][15][16][17][18][19][20] More importantly, a recent study suggests that restoration of dystrophin in skeletal muscles only may exacerbate the failure of heart function if dystrophin expression cannot be effectively restored in cardiac muscle. 16 However, both unmodified 2 0 O methyl phosphorothioate AONs and morpholino oligomers have been unable to induce effective exon skipping and dystrophin expression in cardiac muscle.…”

Section: Introductionmentioning

confidence: 99%

Dose-dependent restoration of dystrophin expression in cardiac muscle of dystrophic mice by systemically delivered morpholino

et al. 2009

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We have earlier shown that antisense morpholino oligomers are able to restore dystrophin expression by systemic delivery in body-wide skeletal muscles of dystrophic mdx mice. However, the levels of dystrophin expression vary considerably and, more importantly, no dystrophin expression has been achieved in cardiac muscle. In this study, we investigate the efficiency of morpholino-induced exon skipping in cardiomyoblasts and myocytes in vitro, and in cardiac muscle in vivo by dose escalation. We showed that morpholino induces targeted exon skipping equally effectively in both skeletal muscle myoblasts and cardiomyoblasts. Effective exon skipping was achieved in cardiomyocytes in culture. In the mdx mice, morpholino rescues dystrophin expression dose dependently in both skeletal and cardiac muscles. Therapeutic levels of dystrophin were achieved in cardiac muscle albeit at higher doses than in skeletal muscles. Up to 50 and 30% normal levels of dystrophin were induced by single systemic delivery of 3 g kg -1 of morpholino in skeletal and cardiac muscles, respectively. High doses of morpholino treatment reduced the serum levels of creatine kinase without clear toxicity. These findings suggest that effective rescue of dystrophin in cardiac muscles can be achieved by morpholino for the treatment of Duchenne muscular dystrophy.

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Visualization of Dystrophic Muscle Fibers in Mdx Mouse by Vital Staining with Evans Blue: Evidence of Apoptosis in Dystrophin-Deficient Muscle

Cited by 332 publications

References 25 publications

Hydroporation as the mechanism of hydrodynamic delivery

Hydroporation as the mechanism of hydrodynamic delivery

Noninvasive monitoring of therapeutic gene transfer in animal models of muscular dystrophies

Dose-dependent restoration of dystrophin expression in cardiac muscle of dystrophic mice by systemically delivered morpholino

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