TY - JOUR
T1 - Measurement of oxygen consumption in mouse aortic endothelial cells using a microparticulate oximetry probe
AU - Pandian, Ramasamy P.
AU - Kutala, Vijay Kumar
AU - Parinandi, Narasimham L.
AU - Zweier, Jay L.
AU - Kuppusamy, Periannan
PY - 2003/12/1
Y1 - 2003/12/1
N2 - The purpose of this study was to determine the rate of oxygen consumption in mouse aortic endothelial cells (MAECs) and to determine the effect of a variety of inhibitors and stimulators of oxygen consumption measured by electron paramagnetic resonance (EPR) spectroscopy utilizing a new particulate oximetry probe. We have previously demonstrated that the octa-n-butoxy derivative of naphthalocyanine neutral radical (LiNc-BuO) enables accurate, precise, and reproducible measurements of pO2 in cellular suspensions. In the current study, we carried out measurements to provide an accurate determination of pO2 in small volume with less number of cells (20,000 cells) that has not been possible with other techniques. To establish the reliability of this method, agents such as menadione, lipopolysaccharide (LPS), potassium cyanide, rotenone, and diphenyleneiodonium chloride (DPI) were used to modulate the oxygen consumption rate in the cells. We observed an increase in oxygen consumption by the cells upon treatment with menadione and LPS, whereas treatment with cyanide, rotenone, and DPI inhibited oxygen consumption. This study clearly demonstrated the utilization of EPR spectrometry with LiNc-BuO probe for determination of oxygen concentration in cultured cells.
AB - The purpose of this study was to determine the rate of oxygen consumption in mouse aortic endothelial cells (MAECs) and to determine the effect of a variety of inhibitors and stimulators of oxygen consumption measured by electron paramagnetic resonance (EPR) spectroscopy utilizing a new particulate oximetry probe. We have previously demonstrated that the octa-n-butoxy derivative of naphthalocyanine neutral radical (LiNc-BuO) enables accurate, precise, and reproducible measurements of pO2 in cellular suspensions. In the current study, we carried out measurements to provide an accurate determination of pO2 in small volume with less number of cells (20,000 cells) that has not been possible with other techniques. To establish the reliability of this method, agents such as menadione, lipopolysaccharide (LPS), potassium cyanide, rotenone, and diphenyleneiodonium chloride (DPI) were used to modulate the oxygen consumption rate in the cells. We observed an increase in oxygen consumption by the cells upon treatment with menadione and LPS, whereas treatment with cyanide, rotenone, and DPI inhibited oxygen consumption. This study clearly demonstrated the utilization of EPR spectrometry with LiNc-BuO probe for determination of oxygen concentration in cultured cells.
KW - Endothelial cells
KW - EPR oximetry
KW - Lipopolysaccharide
KW - Menadione
KW - Oxygen consumption
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U2 - 10.1016/j.abb.2003.09.008
DO - 10.1016/j.abb.2003.09.008
M3 - Article
C2 - 14622987
AN - SCOPUS:0242690343
SN - 0003-9861
VL - 420
SP - 169
EP - 175
JO - Archives of Biochemistry and Biophysics
JF - Archives of Biochemistry and Biophysics
IS - 1
ER -