Measurement of oxygen consumption by murine tissues in vitro
| Autor: | Abdul-Kader Souid, Walter Conca, Farida Marzouqi, Manjusha Sudhadevi, Mohammed T. Al Samri, Mariam Al Shamsi, Harvey S. Penefsky, Shaikha K.M. Al Dawaar, Ruqayya S.M.S. Al Hanjeri, Suleiman Al-Hammadi, Sheela Benedict, Suhail Al-Salam, Aysha Al Mansouri, Ghazala Balhaj |
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| Rok vydání: | 2010 |
| Předmět: |
Male
Pathology medicine.medical_specialty Caspase 3 Biology Toxicology Electron Transport Mice Oxygen Consumption Respiratory Rate Sodium Cyanide Respiration medicine Animals Pharmacology Kidney Mice Inbred BALB C Histology Molecular biology In vitro Staining Mitochondria Respiratory Function Tests Oxygen medicine.anatomical_structure Mitochondrial respiratory chain Apoptosis Energy Metabolism Oxidation-Reduction |
| Zdroj: | Journal of pharmacological and toxicological methods. 63(2) |
| ISSN: | 1873-488X |
| Popis: | Introduction A novel in vitro system was developed to measure O2 consumption by murine tissues over several hours. Methods Tissue specimens (7–35 mg) excised from male Balb/c mice were immediately immersed in ice-cold Krebs–Henseleit buffer, saturated with 95% O2:5% CO2. The specimens were incubated at 37 °C in the buffer, continuously gassed with O2:CO2 (95:5). [O2] was determined as a function of time from the phosphorescence decay rates (1 / τ) of Pd(II) meso-tetra-(4-sulfonatophenyl)-tetrabenzoporphyrin. The values of 1 / τ were linear with [O2]: 1 / τ = 1/τo + kq [O2]; 1 / τo = the decay rate for zero O2, kq = the rate constant in s−1 μM−1. Results NaCN inhibited O2 consumption, confirming oxidation occurred in the mitochondrial respiratory chain. The rate of respiration in lung specimens incubated in vitro for 3.9 ≤ t ≤ 12.4 h was 0.24 ± 0.03 μM O2 min−1 mg−1 (mean ± SD, n = 28). The corresponding rate for the liver was 0.27 ± 0.13 (n = 11, t ≤ 4.7 h), spleen 0.28 ± 0.07 (n = 10, t ≤ 5 h), kidney 0.34 ± 0.12 (n = 7, t ≤ 5 h) and pancreas 0.35 ± 0.09 (n = 10, t ≤ 4 h). Normal tissue histology at hour 5 was confirmed by light and electron microscopy. There was negligible number of apoptotic cells by caspase 3 staining. Discussion This approach allows accurate assessment of tissue bioenergetics in vitro. |
| Databáze: | OpenAIRE |
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