Abstract
The state of reduction of mitochondrial pyridine nucleotides (PN) and flavoproteins (FP) can in principle be estimated in tissue from their fluorescence.1 Although this technique has teen applied quantitatively to isolated mitochondria, there have been major problems in obtaining quantitative estimates of the state of mitochondrial reduction in vivo. Two basic categories of difficulties occur in the application to intact tissue: (1) heterogeneity of the signal due to nonspecific tissue reflectance and/or fluorescence, limited spatial resolution, variable mitochondrial content and interference by extraneous chromophores such as hemoglobin and myoglobin; and (2) lack of reproducible, absolute calibration of the recording apparatus for maximally oxidized and reduced tissue signals. We have developed techniques to overcome some of the above limitations. By using frozen tissue samples and making measurements at liquid N2 temperatures with laser enhanced activation light, greater fluorescence yields are obtained and the spatial resolution is improved.2 The use of the ratio of two indicators, flavoprotein and pyridine nucleotides, helps minimize interference by variable mitochondrial concentrations and the effect of extraneous chromophores.2 Finally, samples corresponding to full oxidation and full reduction can be made available for calibration. This report describes the application of these techniques and other methodological improvements to quantitate the state of mitochondrial redox during the transition from rest to heavy work in dog gracilis muscle.
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References
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Olgin, J., Connect, R.J., Chance, B. (1986). Mitochondrial Redox Changes during Rest-Work Transition in Dog Gracilis Muscle. In: Longmuir, I.S. (eds) Oxygen Transport to Tissue VIII. Advances in Experimental Medicine and Biology, vol 200. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-5188-7_67
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DOI: https://doi.org/10.1007/978-1-4684-5188-7_67
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