Abstract
Histoenzymological methods usually performed on muscle fibres have been adapted to assess the functioning of oxidative phosphorylation in human circulating blood lymphocytes and monocytes. Oxidases and dehydrogenases were analysed in lymphocyte/monocyte smears. The specificity of each histoenzymological reaction was tested using a specific respiratory chain inhibitor: rotenone for NADH diaphorase, thenoyltrifluoroacetone for succinate dehydrogenase, potassium cyanide for cytochrome c oxidase and oligomycin for ATPase. Complex I activity was detected, but inhibition with rotenone was incomplete. Complexes II, IV and V were almost completely inhibited. These observations indicate that histoenzymology is a valuable method for detecting the activity of these oxidative phosphorylation enzymes in lymphocytes and monocytes. The histoenzymology tests performed on fresh peripheral blood cells resembled those used for muscle biopsies. They could be useful for the diagnosis of respiratory chain disorders in patients.
Similar content being viewed by others
References cited
Bancroft JD (1994) Enzyme histochemistry. In: Bancroft JD, Stevens A, eds. Theory and Practice of Histological Techniques. 3rd Edn., Edinburgh: Churchill Livingstone, pp. 379-399.
Barroso N, Campos Y, Huertas R, Esteban J, Molina A, Alonso A, Gutierrez-Rivas E, Arenas J (1993) Respiratory chain enzyme activities in lymphocytes from untreated patients with Parkinson's disease. Clin Chem 39: 667-669.
Bojes HK, Sausen PJ, Cattley RC, Keller BJ, Thurman RG (1996) Paradoxical increase in peroxisomal cyanide-insensitive respiration following dietary exposure to WY-14,643 in the perfused liver. Toxicol Appl Pharmacol 137: 202-209.
Chretien D, Rustin P, Bougeron T, Rotig A, Saudubray JM, Munnich A (1994) Reference charts for respiratory chain activities in human tissues. Clin Chim Acta 228: 53-70.
Cooper HL (1974) Purification of lymphocytes from peripheral blood. Methods Enzymol XXXII: 633-637.
Dubowitz V, Secury CA, Fitzsimans RB (1985) Muscle Biopsy: A Practical Approach. London: Baillière Tindall & W. Saunders.
Kriege D, Carroll MT, Cooper JM, Marsden FRS, Schapira AHV (1992) Platelet mitochondrial function in Parkinson's disease. Ann Neurol 32: 782-788.
Letellier T, Malgat M, Coquet M, Moretto B, Parrot-Rouland F, Mazat JP (1992) Mitochondrial myopathy studies on permeabilized muscle fibers. Pediat Res 32: 17-22.
Luft R (1994) The development of mitochondrial medicine. Proc Natl Acad Sci USA 91: 8731-8738.
Martin MA, Molina JA, Jimenez-Jimenez FJ, Benito-León J, Ortí-Pareja M, Campos Y, Arenas J (1996) Respiratory-chain enzyme activities in isolated mitochondria of lymphocytes from untreated Parkinson's disease patients. Neurology 46: 1343-1346.
Meijer AEFH, Vloedman AHT (1980) The histochemical characterisation of the coupling state of skeletal muscle mitochondria. Histochemistry 69: 217-232.
Molina JA, De Bustos F, Jimenez-Jimenez FJ, Benito Leon J, Gasalla T, Orti-Pareja M, Vela L, Bermejo F, Martin MA, Campos Y, Arenas J (1997) Respiratory chain enzyme activities in isolated mitochondria of lymphocytes from patients with Alzheimer's disease. Neurology 48: 636-638.
Munnich A, Rötig A, Chretien D, CarmierV, Bourgeron T, Bonnefont JP, Saudubray JM, Rustin P (1996) Clinical presentation of mitochondrial disorders in childhood. J Inher Metab Dis 19: 521-527.
Olivera AA, Meigs RA (1975) Mitochondria from human term placenta. II. Characterization of respiratory pathways and coupling mechanisms. Biochim Byophis Acta 376: 436-445.
Parker WD, Filley ChM, Parks JK (1990) Cytochrome oxidase deficiency in Alzheimer's disease. Neurology 40: 1302-1303.
Romero NB, Lombés A, Touati G, Rigal O, Frachon P, Cheval MA, Giraud M, Possekel S, Fardeau M, Ogier De Baulny H (1996) Morphological studies of skeletal muscle in lactic acidosis. J Inher Metab Dis 19: 528-534.
Rowland LP (1994) Mitochondrial encephalomyopathies: lumping, splitting and melding. In: Schapira AVH, DiMauro S, eds. Mitochondrial Disorders in Neurology. Oxford: Butterworth-Heinemann, pp. 116-129.
Rustin P, Chretien D, Bourgeron T, Gerard B, Rotig A, Munich A (1994) Biochemical and molecular investigation in respiratory chain deficiencies. Clin Chim Acta 228: 35-51.
Seligman AM, Karnowsky MJ, Wasserkrug HL, Hanker JS (1968) Nondroplet ultrastructural demonstration of cytochrome oxidase activity with a polymerizing osmiophilic reagent, diaminobenzidine (DAB). J Cell Biol 38: 1-14.
Shoffner JM, Wallace DC (1995) Oxidative phosphorylation disease. In: Scriver Ch R, Boudet AL, Sly WS, Valle D, eds. The Metabolic and Molecular Basis of Inherited Disease. 7th Edn., New York: Mc Graw-Hill Inc., pp. 1535-1610.
Skegg PD (1977) English law relating to experimentation on children. Lancet 2: 754-755.
Stevens A (1994) Enzyme Histochemistry: Diagnostic Applications in Theory and Practice of Histological Techniques. Bancroft JD, Stevens A, eds, 3rd Edn., Edinburgh: Churchill Livingstone pp. 401-411.
Trounce IA, Kim YL, Jun AS, Wallace D (1996) Assessment of mitochondrial oxidative phosphorylation in patient muscle biopsies, lymphoblasts and transmitochondrial cell lines. Meth Enzymol 264: 487-508.
Van Zuylen AJ, Basman GJCGM, Ruitenbeek W, Van Kalmthout PJC, De Grip WJ (1992) No evidence for reduced thrombocyte cytochrome oxidase activity in Alzheimer's disease. Neurology 42: 1246-1247.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
de Halac, I.N., Bacman, S.R. & de Kremer, R.D. Histoenzymology of Oxidases and Dehydrogenases in Peripheral Blood Lymphocytes and Monocytes for the Study of Mitochondrial Oxidative Phosphorylation. Histochem J 32, 133–137 (2000). https://doi.org/10.1023/A:1004078705513
Issue Date:
DOI: https://doi.org/10.1023/A:1004078705513