Immunocytochemical studies of cytochrome oxidase subunits in skeletal muscle of patients with partial cytochrome oxidase deficiencies

Johnson, Margaret; Kadenbach, Bernhard; Droste, M.; Old, Sally L.; Turnbull, Douglass M.

doi:10.1016/0022-510x(88)90056-1

Cited by 49 publications

(12 citation statements)

References 23 publications

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“…In our laboratory, we carry out the study with unfi xed frozen sections, but with some antibodies, permeabilization of the mitochondrial membranes may be required to uncover the antigenetic sequences or to facilitate the penetration of the probes into the inner mitochondrial compartment. In agreement with Johnson et al ( 41 ) , using 4% formaldehyde in 0.1 M CaCl 2 , pH 7, followed by dehydration provides good results (see Subheading 3.2.3 ).…”

Section: Notessupporting

confidence: 86%

Morphological Assessment of Mitochondrial Respiratory Chain Function on Tissue Sections

Tanji

2011

Methods in Molecular Biology

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In recent decades, genetic, biochemical, immunological, and cell biological techniques have been applied not only for better understanding of pathogenesis of known mitochondrial encephalomyopathies but also for exploring the possibility of mitochondrial involvement in other neurological, cardiac, gastrointestinal, and urological diseases. Techniques applied in a coordinated fashion have made it clear that mitochondrial dysfunction plays an important role in a wide range of human diseases including degenerative, toxic, metabolic, and neoplastic disorders. In this chapter, we provide updated protocols of essential histochemical and immunohistochemical methods that, in our opinion, are the most reliable morphological tool to visualize respiratory chain abnormality on tissue sections.

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Section: Notessupporting

confidence: 86%

Morphological Assessment of Mitochondrial Respiratory Chain Function on Tissue Sections

Tanji

2011

Methods in Molecular Biology

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“…In some cytochrome oxidase deficiencies the defect is confined to a subpopulation of muscle fibres (Johnson et al, 1983) and assays made on muscle homogenates may fail to detect any abnormality, especially in the early stages of the disorder (Sherratt et al, 1988). The ability to make enzyme determinations in single muscle fibres is invaluable in this respect (Reichmann, 1988) and results can be correlated with immunocytochemical data on enzyme subunit proteins in the same individual fibres (Johnson et al, 1988). These techniques provide useful ways of investigating respiratory complexes at the cellular level.…”

Section: Discussionmentioning

confidence: 98%

Methods of microphotometric assay of succinate dehydrogenase and cytochromec oxidase activities for use on human skeletal muscle

Old¹,

Johnson²

1989

Histochem J

171

120

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Microphotometric assay media for the measurement of succinate dehydrogenase (SDH) and cytochrome oxidase activities in sections of human skeletal muscle have been developed. The optimal constitution of these media was determined experimentally. Factors investigated include the effects of substrate concentration, pH, use of different electron acceptors and electron donors, influence of intermediate electron carriers and tissue-stabilizing agents, effects of inhibitors, the extent of endogenous and non-specific reactions and the linearity of the reactions during the time course of the assays. Optimal assay media (SDH) contained 130 mM succinate, 1.5 mM Nitro Blue tetrazolium, 0.2 mM phenazine methosulphate and 1.0 mM sodium azide in 0.1 m phosphate buffer, pH7.0. Cytochrome oxidase was optimally assayed in media containing 4 mM diaminobenzidine and 100 microns cytochrome c. Reactions in individual muscle fibers were found to be linear for incubation times up to 10 min in SDH assays and for more than 15 min in cytochrome oxidase determinations. Some potential uses of these microphotometric assays in the investigation of human metabolic muscle disorders are discussed.

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“…always accompanied by parallel changes in mitochondrial subunit proteins, while changes in nuclear ones are much less severe and are variable (Johnson et al, 1988;Mita et al, 1989). At the transcriptional level, found that cytochrome b mRNA (mitochondrial) increased three-fold more than CO VIc mRNA (nuclear) after 21 days of muscle stimulation.…”

Section: Co Vbmentioning

confidence: 96%

“…We cannot rule out whether the regulation of each individual CO subunit follows the same mechanism. For example, different nuclear-encoded subunits might be differentially regulated, since they displayed a great variety of change in their proteins (Johnson et al, 1988;Liu and Wong-Riley, 1995) and mRNAs (Williams and Harlen, 1987;William et al, 1987;Hevner and WongRiley, 1993). One of the ways to probe this issue further is to compare quantitatively the time course and degree of change in each subunit proteins and their mRNAs in this experimental system.…”

Section: Co Vbmentioning

confidence: 97%

Mitochondrial- and nuclear-encoded subunits of cytochrome oxidase in neurons: Differences in compartmental distribution, correlation with enzyme activity, and regulation by neuronal activity

Nie

Wong‐Riley

1996

J. Comp. Neurol.

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Cytochrome oxidase (CO), a mitochondrial energy-generating enzyme, contains both mitochondrial- and nuclear-encoded subunits. In neurons, local levels of CO activity vary among different neuronal compartments, reflecting local demands for energy. The goals of the present study were to determine if compartmental distribution of CO subunit proteins from the two genomes was correlated with local CO activity, and if their expression was regulated proportionately in neurons. The subcellular distributions of mitochondrial-encoded CO III and nuclear-encoded CO Vb proteins were quantitatively analyzed in mouse cerebellar sections subjected to postembedding immunocytochemistry. Local levels of subunit proteins were also compared to local CO activity, as revealed by CO cytochemistry. In order to study the regulation of subunit protein expression, we assessed changes in immunoreactivity of the two CO subunits as well as changes in CO activity in mouse superior colliculus after 1 to 7 days of monocular enucleation. We found that immunoreaction product for both CO III and CO Vb existed almost exclusively in mitochondria, but their compartmental distributions were different. CO III was nonhomogeneously distributed among different neuronal compartments, where its local level was positively correlated with that of CO activity. In contrast, the subcellular distribution of CO Vb was relatively uniform and did not bear a direct relationship with that of CO activity. Moreover, the two subunit proteins were disproportionately regulated by neuronal activity. CO III and CO activity exhibited parallel decreases after the deprivation of afferent input, and their changes were earlier and to a greater degree than that of CO Vb proteins. Thus, the present findings indicate that the local expression and/or distribution of CO subunit proteins from the two genomes may involve different regulatory mechanisms in neurons. Our data also suggest that the activity-dependent regulation of mitochondrial-encoded CO subunits is likely to play a major role in controlling the local levels of CO content and its activity.

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Immunocytochemical studies of cytochrome oxidase subunits in skeletal muscle of patients with partial cytochrome oxidase deficiencies

Cited by 49 publications

References 23 publications

Morphological Assessment of Mitochondrial Respiratory Chain Function on Tissue Sections

Morphological Assessment of Mitochondrial Respiratory Chain Function on Tissue Sections

Methods of microphotometric assay of succinate dehydrogenase and cytochromec oxidase activities for use on human skeletal muscle

Mitochondrial- and nuclear-encoded subunits of cytochrome oxidase in neurons: Differences in compartmental distribution, correlation with enzyme activity, and regulation by neuronal activity

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