In situ Raman study of redox state changes of mitochondrial cytochromes in a perfused rat heart

TY - JOUR

T1 - In situ Raman study of redox state changes of mitochondrial cytochromes in a perfused rat heart

AU - Brazhe, Nadezda

AU - Treiman, Marek

AU - Faricelli, Barbara

AU - Vestergaard, Jakob Hedemark

AU - Sosnovtseva, Olga

PY - 2013/8/29

Y1 - 2013/8/29

N2 - We developed a Raman spectroscopy-based approach for simultaneous study of redox changes in c-and b-type cytochromes and for a semiquantitative estimation of the amount of oxygenated myoglobin in a perfused rat heart. Excitation at 532 nm was used to obtain Raman scattering of the myocardial surface of the isolated heart at normal and hypoxic conditions. Raman spectra of the heart under normal pO2 demonstrate unique peaks attributable to reduced c-and b-type cytochromes and oxymyoglobin (oMb). The cytochrome peaks decreased in intensity upon FCCP treatment, as predicted from uncoupling mitochondrial respiration. Conversely, transient hypoxia causes the reversible increase in the intensity of peaks assigned to cytochromes c and c1, reflecting electron stacking proximal to cytochrome oxidase due to the lack of terminal electron acceptor O2. Intensities of peaks assigned to oxy- and deoxyhemoglobin were used for the semiquantitative estimation of oMb deoxygenation that was found to be of approximately 50% under hypoxia conditions.

AB - We developed a Raman spectroscopy-based approach for simultaneous study of redox changes in c-and b-type cytochromes and for a semiquantitative estimation of the amount of oxygenated myoglobin in a perfused rat heart. Excitation at 532 nm was used to obtain Raman scattering of the myocardial surface of the isolated heart at normal and hypoxic conditions. Raman spectra of the heart under normal pO2 demonstrate unique peaks attributable to reduced c-and b-type cytochromes and oxymyoglobin (oMb). The cytochrome peaks decreased in intensity upon FCCP treatment, as predicted from uncoupling mitochondrial respiration. Conversely, transient hypoxia causes the reversible increase in the intensity of peaks assigned to cytochromes c and c1, reflecting electron stacking proximal to cytochrome oxidase due to the lack of terminal electron acceptor O2. Intensities of peaks assigned to oxy- and deoxyhemoglobin were used for the semiquantitative estimation of oMb deoxygenation that was found to be of approximately 50% under hypoxia conditions.

U2 - 10.1371/journal.pone.0070488

DO - 10.1371/journal.pone.0070488

M3 - Journal article

C2 - 24009655

SN - 1932-6203

VL - 8

SP - e70488

JO - PLoS Computational Biology

JF - PLoS Computational Biology

IS - 8

ER -