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Biochemical evaluation of mitochondrial respiratory chain enzymes in canine skeletal muscle

Joseph J. WakshlagDepartment of Biomedical Science and Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14850.

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 DVM, MS
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Barry J. CooperDepartment of Biomedical Science and Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14850.

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 BVSc, PhD
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Robin R. WakshlagDepartment of Biomedical Science and Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14850.

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Francis. A. KallfelzDepartment of Biomedical Science and Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14850.

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Stephen C. BarrDepartment of Biomedical Science and Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14850.

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Daryl V. NydamDepartment of Biomedical Science and Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14850.

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S. DimauroDepartment of Neurology, Columbia College of Physicians and Surgeons, New York City, NY 10018.

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 PhD

Abstract

Objective—To perform respiratory chain enzymatic activity assays on canine skeletal muscle biopsy specimens and establish reference range values of skeletal muscle enzyme activities for dogs.

Sample Population—Biopsy specimens from the vastus lateralis muscle were obtained from 24 dogs (8 sexually intact males and 14 sexually intact females) ranging from 15 months to 6 years of age.

Procedure—Mean values of citrate synthase, cytochrome-c oxidase, succinate dehydrogenase, succinate dehydrogenase-cytochrome-c reductase, nicotinamide adenine dinucleotide (NADH) dehydrogenase, and NADH dehydrogenase-cytochrome-c reductase activities were established by use of 6 standard spectrophotometric assays for respiratory chain enzyme analysis.

Results—Compared with published data for skeletal muscle enzyme activities in humans, skeletal muscle enzyme activities in dogs were 2- to 4-fold higher. Additionally, citrate synthase activity, a marker for mitochondrial volume, was positively correlated with age in dogs, suggesting that mitochondrial volume increases with age, although no apparent change in respiratory chain enzymatic activity with an increase in age was found.

Conclusions and Clinical Relevance—Reference range values for skeletal muscle enzyme activities of dogs are needed to accurately interpret results of respiratory chain enzymatic activity assays. During investigation of metabolic myopathies, if skeletal muscle biopsy specimens are evaluated for respiratory chain enzyme kinetics, they should be performed and evaluated in concert with skeletal muscle biopsy specimens from clinically normal animals of the same species. (Am J Vet Res 2004;65:480–484)

Abstract

Objective—To perform respiratory chain enzymatic activity assays on canine skeletal muscle biopsy specimens and establish reference range values of skeletal muscle enzyme activities for dogs.

Sample Population—Biopsy specimens from the vastus lateralis muscle were obtained from 24 dogs (8 sexually intact males and 14 sexually intact females) ranging from 15 months to 6 years of age.

Procedure—Mean values of citrate synthase, cytochrome-c oxidase, succinate dehydrogenase, succinate dehydrogenase-cytochrome-c reductase, nicotinamide adenine dinucleotide (NADH) dehydrogenase, and NADH dehydrogenase-cytochrome-c reductase activities were established by use of 6 standard spectrophotometric assays for respiratory chain enzyme analysis.

Results—Compared with published data for skeletal muscle enzyme activities in humans, skeletal muscle enzyme activities in dogs were 2- to 4-fold higher. Additionally, citrate synthase activity, a marker for mitochondrial volume, was positively correlated with age in dogs, suggesting that mitochondrial volume increases with age, although no apparent change in respiratory chain enzymatic activity with an increase in age was found.

Conclusions and Clinical Relevance—Reference range values for skeletal muscle enzyme activities of dogs are needed to accurately interpret results of respiratory chain enzymatic activity assays. During investigation of metabolic myopathies, if skeletal muscle biopsy specimens are evaluated for respiratory chain enzyme kinetics, they should be performed and evaluated in concert with skeletal muscle biopsy specimens from clinically normal animals of the same species. (Am J Vet Res 2004;65:480–484)