Glutathione depletion during experimental damage to rat skeletal muscle and its relevance to Duchenne muscular dystrophy

1. The release of glutathione has been studied in comparison with the release of creatine kinase from isolated rat soleus muscles subjected to certain forms of experimental damage. 2. Excessive electrically stimulated contractile activity or treatment of muscles with the mitochondrial inhibitor, 2,4-dinitrophenol, induced a substantial release of both creatine kinase and glutathione and a reduction in the total glutathione content of the muscle. The time course of this release and depletion indicates that the efflux of the two molecules is not directly related and that a reduction in muscle glutathione content does not occur before cytosolic enzyme release. 3. 2,4-Dinitrophenol-stimulated release of creatine kinase was significantly reduced by the omission of external calcium from the incubation media, but glutathione release and depletion was relatively unaffected by this. Deliberate elevation of the muscle intracellular calcium content with the calcium ionophore, A23187, induced a substantial loss of creatine kinase, but had no significant effect on the release of glutathione. 4. Muscle biopsies from patients with Duchenne muscular dystrophy were found to have an elevated content of glutathione and an equivalent protein-thiol content compared with control subjects. 5. We conclude that, although release of glutathione from skeletal muscle occurs after excessive contractile activity or inhibition of mitochondrial metabolism, this is not a key step in the damaging processes leading to cytosolic enzyme release, neither is it relevant to the ongoing damage to skeletal muscle which occurs in patients with Duchenne muscular dystrophy.

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Release of creatine kinase and prostaglandin E2 from regenerating skeletal muscle fibers

Journal of Applied Physiology ◽

10.1152/jappl.1994.76.3.1274 ◽

1994 ◽

Vol 76 (3) ◽

pp. 1274-1278 ◽

Cited By ~ 15

Author(s):

A. McArdle ◽

R. H. Edwards ◽

M. J. Jackson

Keyword(s):

Skeletal Muscle ◽

Creatine Kinase ◽

Prostaglandin E2 ◽

Extensor Digitorum Longus ◽

Contractile Activity ◽

Calcium Ionophore ◽

Sectional Area ◽

Cross Sectional ◽

A 23187

To study the effect of regeneration on the release of creatine kinase (CK) and prostaglandin E2 from muscle, extensor digitorum longus muscles of mice were injected with 50 microliters of BaCl2 solution in saline (1.2% wt/vol). Injected muscles showed almost complete degeneration at 3 days postinjection but had regenerated to approximately the same fiber cross-sectional area as contralateral control muscles by 12 days postinjection. These muscles released reduced amounts of intracellular CK compared with contralateral control muscles in response to excessive isometric contractile activity or treatment with the calcium ionophore A-23187 (20 microM) in vitro. However, regenerating muscles contained a lower total CK activity than contralateral control muscles, which accounted for the reduced CK efflux after experimental damage. Regenerating muscles released an increased amount of prostaglandin E2 compared with control muscles after both damaging stimuli. We conclude that regenerated muscles show no reduced susceptibility to contraction-induced damage (as assessed by CK release) but show an elevated release of prostaglandin E2 in response to contractile activity or an increase in intracellular calcium.

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Single-Blind Study of Dystrophin Staining in Carriers of Duchenne Muscular Dystrophy

Canadian Journal of Neurological Sciences / Journal Canadien des Sciences Neurologiques ◽

10.1017/s0317167100047399 ◽

1993 ◽

Vol 20 (1) ◽

pp. 44-47 ◽

Cited By ~ 6

Author(s):

Francois P. Bernier ◽

Cheryl R. Greenberg ◽

William C. Halliday ◽

Klaus Wrogemann

Keyword(s):

Skeletal Muscle ◽

Creatine Kinase ◽

Duchenne Muscular Dystrophy ◽

Muscular Dystrophy ◽

Diagnostic Test ◽

Family Members ◽

Immunohistochemical Analysis ◽

Blind Study ◽

Single Blind

ABSTRACT:A single-blind study of dystrophin staining in skeletal muscle was performed in 13 biopsies from carriers of Duchenne Muscular Dystrophy (DMD) and controls. The results indicate that immunohistochemical analysis of dystrophin staining is a valuable diagnostic test for DMD carriers when DNA for testing is unavailable from critical family members or is uninformative, when creatine kinase (CK) values are conflicting or when CK values must be used in isolation.

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Duchenne muscular dystrophy: 45Ca Exchange in cultured skin fibroblasts and the effect of calcium ionophore A23187

Clinica Chimica Acta ◽

10.1016/0009-8981(79)90432-7 ◽

1979 ◽

Vol 96 (3) ◽

pp. 225-231 ◽

Cited By ~ 5

Author(s):

Helen E. Statham ◽

Victor Dubowitz

Keyword(s):

Duchenne Muscular Dystrophy ◽

Muscular Dystrophy ◽

Calcium Ionophore ◽

Calcium Ionophore A23187 ◽

Skin Fibroblasts ◽

Ionophore A23187 ◽

Cultured Skin

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Effect of inhibitors of arachidonic acid metabolism on efflux of intracellular enzymes from skeletal muscle following experimental damage

Biochemical Journal ◽

10.1042/bj2410403 ◽

1987 ◽

Vol 241 (2) ◽

pp. 403-407 ◽

Cited By ~ 39

Author(s):

M J Jackson ◽

A J M Wagenmakers ◽

R H T Edwards

Keyword(s):

Skeletal Muscle ◽

Creatine Kinase ◽

Arachidonic Acid ◽

Acid Metabolism ◽

Membrane Integrity ◽

Calcium Ionophore ◽

Arachidonic Acid Metabolism ◽

Ionophore A23187 ◽

Intracellular Enzymes

The role of arachidonic acid metabolism in the efflux of intracellular enzymes from damaged skeletal muscle has been examined in vitro using inhibitors of cyclo-oxygenase and lipoxygenase enzymes. Damage to skeletal muscle induced by either calcium ionophore A23187 (25 microM) or dinitrophenol (1 mM) caused an increase in the efflux of prostaglandins E2 and F2 alpha together with a large efflux of intracellular creatine kinase. Use of a cyclo-oxygenase inhibitor completely prevented the efflux of prostaglandins, but had no effect on creatine kinase efflux. However, several agents having the ability to inhibit lipoxygenase enzymes dramatically reduced creatine kinase efflux following damage. These data suggest that a product or products of lipoxygenase enzymes may be mediators of the changes in plasma membrane integrity which permit efflux of intracellular enzymes as a consequence of skeletal muscle damage.

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Isoenzyme distribution of creatine kinase and lactate dehydrogenase in serum and skeletal muscle in Duchenne muscular dystrophy, collagen disease, and other muscular disorders.

Clinical Chemistry ◽

10.1093/clinchem/24.11.1985 ◽

1978 ◽

Vol 24 (11) ◽

pp. 1985-1989 ◽

Cited By ~ 44

Author(s):

W G Yasmineh ◽

G A Ibrahim ◽

M Abbasnezhad ◽

E A Awad

Keyword(s):

Skeletal Muscle ◽

Creatine Kinase ◽

Lactate Dehydrogenase ◽

Duchenne Muscular Dystrophy ◽

Muscular Dystrophy ◽

Serum Creatine Kinase ◽

Collagen Disease ◽

Total Serum ◽

Control Group ◽

The Mean

Abstract We determined the total activity and isoenzyme distribution of lactate dehydrogenase and creatine kinase in serum and biopsy specimens from skeletal muscle of nine normal individuals and nine patients with Duchenne muscular dystrophy (I), five with collagen disease (II), and four with non-progressive unclassified myopathy (III). Mean total serum creatine kinase in patients with Duchenne muscular dystrophy (867 U/liter, SD = 197) was 31-fold that in the control group (28 U/liter, SD = 14). There was also a small (3.3-fold) increase in the mean total serum creatine kinase of patients with III, but none in the serum from patients with II. Changes in the creatine kinase isoenzyme distribution of skeletal muscle were primarily in the MB isoenzyme. The mean percentage of creatine kinase-MB activity in muscle from patients with I (2.81, SD = 1.15) and patients with III (1.69, SD = 1.07) significantly (P less than 0.005) exceeded that of the control group (0.43, SD = 0.18). Muscle from patients with II showed little change. The most striking changes in lactate dehydrogenase were also observed in patients with I, in whom the mean total serum activity (356 U/liter, SD = 115) was 3.4-fold that of serum from the control group (105 U/liter, SD = 19). Skeletal muscle from these patients also showed a significant decrease in mean percent isoenzyme 5 activity (from 50 to 23) and an increase in that of isoenzymes 1 and 2 (from 1 to 9 and 8 to 20, respectively). These changes in the distribution of these two sets of isoenzymes in muscle were reflected in the serum.

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Characterization of a Blood Spot Creatine Kinase Skeletal Muscle Isoform Immunoassay for High-Throughput Newborn Screening of Duchenne Muscular Dystrophy

Clinical Chemistry ◽

10.1373/clinchem.2016.268425 ◽

2017 ◽

Vol 63 (4) ◽

pp. 908-914 ◽

Cited By ~ 17

Author(s):

Stuart J Moat ◽

Teemu Korpimäki ◽

Petra Furu ◽

Harri Hakala ◽

Hanna Polari ◽

...

Keyword(s):

Skeletal Muscle ◽

Creatine Kinase ◽

Duchenne Muscular Dystrophy ◽

Muscular Dystrophy ◽

Newborn Screening ◽

High Throughput ◽

Screening Programs ◽

Blood Spots ◽

The Mean ◽

Blood Spot

Abstract BACKGROUND Duchenne muscular dystrophy (DMD) is a progressive, lethal X-linked neuromuscular disorder with an average worldwide incidence of 1:5000. Blood spot creatine kinase (CK) enzyme assays previously used in newborn screening programs for DMD are nonspecific because measured CK enzyme activity is attributable to 3 isoenzyme forms of CK (CK-MM, CK-MB, and CK-BB) and it is the CK-MM isoform that is found predominantly in skeletal muscle. CK-MM is increased in boys with DMD owing to muscle damage. We describe a sensitive and specific automated immunoassay for CK-MM to screen for DMD in blood spots. METHODS The prototype assay was developed on the PerkinElmer GSP® analyzer to enable high-throughput screening. CK-MM was assayed using a solid phase, 2-site immunofluorometric system. Purified human CK-MM was used to create calibrators and controls. RESULTS The limit of blank (LOB), detection (LOD), and quantification (LOQ) values were <1, 3, and 8 ng/mL, respectively. The analytical measurement range was 4–8840 ng/mL. Interassay (n = 40) imprecision was <7% across the analytical range. Cross-reactivity was <5% for CK-MB and 0% for CK-BB. The mean recovery of CK-MM was 101% (range 87%–111%). Blood spots from newborn infants (n = 277) had a mean CK-MM concentration of 155 ng/mL and a 99th centile of 563 ng/mL. The mean blood spot CK-MM concentration from 10 cases of DMD was 5458 ng/mL (range 1217–9917 ng/mL). CONCLUSIONS CK-MM can be reliably quantified in blood spots. The development of this CK-MM assay on a commercial immunoassay analyzer would enable standardized and high-throughput newborn blood spot screening of DMD.

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Faculty Opinions recommendation of microRNA-206 promotes skeletal muscle regeneration and delays progression of Duchenne muscular dystrophy in mice.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.717397850.793153110 ◽

2012 ◽

Author(s):

Michael Rudnicki ◽

Alessandra Pasut

Keyword(s):

Skeletal Muscle ◽

Duchenne Muscular Dystrophy ◽

Muscular Dystrophy ◽

Muscle Regeneration ◽

Skeletal Muscle Regeneration

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Autophagy in the heart is enhanced and independent of disease progression in mus musculus dystrophinopathy models

JRSM Cardiovascular Disease ◽

10.1177/2048004019879581 ◽

2019 ◽

Vol 8 ◽

pp. 204800401987958

Author(s):

HR Spaulding ◽

C Ballmann ◽

JC Quindry ◽

MB Hudson ◽

JT Selsby

Keyword(s):

Skeletal Muscle ◽

Duchenne Muscular Dystrophy ◽

Muscular Dystrophy ◽

Disease Progression ◽

Gene Mutations ◽

Dystrophin Gene ◽

Mdx Mice ◽

Dystrophin Deficiency ◽

Muscle Wasting Disease ◽

Dystrophin Protein

Background Duchenne muscular dystrophy is a muscle wasting disease caused by dystrophin gene mutations resulting in dysfunctional dystrophin protein. Autophagy, a proteolytic process, is impaired in dystrophic skeletal muscle though little is known about the effect of dystrophin deficiency on autophagy in cardiac muscle. We hypothesized that with disease progression autophagy would become increasingly dysfunctional based upon indirect autophagic markers. Methods Markers of autophagy were measured by western blot in 7-week-old and 17-month-old control (C57) and dystrophic (mdx) hearts. Results Counter to our hypothesis, markers of autophagy were similar between groups. Given these surprising results, two independent experiments were conducted using 14-month-old mdx mice or 10-month-old mdx/Utrn± mice, a more severe model of Duchenne muscular dystrophy. Data from these animals suggest increased autophagosome degradation. Conclusion Together these data suggest that autophagy is not impaired in the dystrophic myocardium as it is in dystrophic skeletal muscle and that disease progression and related injury is independent of autophagic dysfunction.

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