Measurement of oxygen consumption by murine tissues in vitro

Mohammed T. Al Samri, Mariam Al Shamsi, Suhail Al-Salam, Farida Marzouqi, Aysha Al Mansouri, Suleiman Al-Hammadi, Ghazala Balhaj, Shaikha K.M. Al Dawaar, Ruqayya S.M.S. Al Hanjeri, Sheela Benedict, Manjusha Sudhadevi, Walter Conca, Harvey S. Penefsky, Abdul Kader Souid

Research output: Contribution to journalArticlepeer-review

17 Citations (Scopus)

Abstract

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]; 1/7τo=the decay rate for zero O2, kq=the rate constant in sμ1τMτ1. Results: NaCN inhibited O2 consumption, confirming oxidation occurred in the mitochondrial respiratory chain. The rate of respiration in lung specimens incubated in vitro for 3.912.4 h was 0.24±0.03±M 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<5h), kidney 0.34±0.12 (n=7, t<5h) and pancreas 0.35±0.09 (n=10, t<4h). 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.

Original languageEnglish
Pages (from-to)196-204
Number of pages9
JournalJournal of Pharmacological and Toxicological Methods
Volume63
Issue number2
DOIs
Publication statusPublished - Mar 2011

Keywords

  • Bioenergetics
  • Cytotoxicity
  • In vitro

ASJC Scopus subject areas

  • Toxicology
  • Pharmacology

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