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(Referência obtida automaticamente do SciELO, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Evaluation by blue native polyacrylamide electrophoresis colorimetric staining of the effects of physical exercise on the activities of mitochondrial complexes in rat muscle

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Autor(es):
A.M. Molnar [1] ; A.A. Alves [2] ; L. Pereira-da-Silva [3] ; D.V. Macedo [4] ; F. Dabbeni-Sala [5]
Número total de Autores: 5
Afiliação do(s) autor(es):
[1] Universidade Estadual de Campinas. Instituto de Biologia. Departamento de Bioquímica - Brasil
[2] Universidade Estadual de Campinas. Instituto de Biologia. Departamento de Bioquímica - Brasil
[3] Universidade Estadual de Campinas. Instituto de Biologia. Departamento de Bioquímica - Brasil
[4] Universidade Estadual de Campinas. Instituto de Biologia. Departamento de Bioquímica - Brasil
[5] University of Padova. Department of Pharmacology - Itália
Número total de Afiliações: 5
Tipo de documento: Artigo Científico
Fonte: Brazilian Journal of Medical and Biological Research; v. 37, n. 7, p. 939-947, 2004-07-00.
Resumo

Blue native polyacrylamide electrophoresis (BN-PAGE) is a technique developed for the analysis of membrane complexes. Combined with histochemical staining, it permits the analysis and quantification of the activities of mitochondrial oxidative phosphorylation enzymes using whole muscle homogenates, without the need to isolate muscle mitochondria. Mitochondrial complex activities were measured by emerging gels in a solution containing all specific substrates for NADH dehydrogenase and cytochrome c oxidase enzymes (complexes I and IV, respectively) and the colored bands obtained were measured by optique densitometry. The objective of the present study was the application of BN-PAGE colorimetric staining for enzymatic characterization of mitochondrial complexes I and IV in rat muscles with different morphological and biochemical properties. We also investigated these activities at different times after acute exercise of rat soleus muscle. Although having fewer mitochondria than oxidative muscles, white gastrocnemius muscle presented a significantly higher activity (26.7 ± 9.5) in terms of complex I/V ratio compared to the red gastrocnemius (3.8 ± 0.65, P < 0.05) and soleus (9.8 ± 0.9, P < 0.001) muscles. Furthermore, the complex IV/V ratio of white gastrocnemius muscle was always significantly higher when compared to the other muscles. Ninety-five minutes of exhaustive physical exercise induced a decrease in complex I/V and complex IV/V ratios after all resting times (0, 3 and 6 h) compared to control (P < 0.05), probably reflecting the oxidative damage due to increasing free radical production in mitochondria. These results demonstrate the possible and useful application of BN-PAGE-histochemical staining to physical exercise studies. (AU)

Processo FAPESP: 00/07962-2 - Da pesquisa básica ao desenvolvimento de uma nova tecnologia de avaliação física
Beneficiário:Denise Vaz de Macedo
Modalidade de apoio: Auxílio à Pesquisa - Regular