Cell lines

P3 cells

Preparation Method

Oxygen consumption was measured at 37 ℃ in cell culture medium. The oxygen consumption rate was measured under 3 different conditions: phosphorylating state (endogenous respiratory condition), non-phosphorylating state with addition of oligomycin A (50 ng/mL), and uncoupled state by the successive addition of carbonyl cyanide m-chlorophenyl hydrazone (CCCP 0.5 μM) to reach the maximal respiration.

Reaction Conditions

50 ng/mL for 3days

Applications

Additional oxygen consumption analysis was performed using the Oroboros oxygraph methodology and showed no difference in endogenous oxygen consumption rate at oligomycin A or CCCP, even with a long-term DCA treatment (3 days)

Animal models

Wt and Tg mice

Preparation Method

Oligomycin A (1mg/kg ) was given by i.v. for 2h before lipopolysaccharide (LPS) and galactosamine (GalN) treatment

Dosage form

intravenous injection (i.v.), 1mg/kg,

Applications

Wt mice treated with oligomycin A have decreased ATP levels in the liver. Concomitantly, these mice showed an enhanced apoptosis in hepatocytes as Tg mice after LPS/GalN administration.

Oligomycin A is an inhibitor of ATP synthase, which inhibits the process taking place on mitochondria coupling membrane that depended on ATP and oxidative phosphorylation^[1].

Oligomycin A produced a concentration-dependent block of Icrac with similar characteristics, but with lower potency than oligomycin B. Fits of averaged concentration-response data to a logistic function yielded IC50 values and slope factor coefficients (respectively) of 13.5 μM and 0.85 for oligomycin A, and 2.3 μM and 0.82 for oligomycin B. A two-way analysis of variance confirmed a significantly greater inhibition by oligomycin B over the concentration range tested^[2].

Oligomycin A treated Wt(widetype) mice have decreased ATP levels in the liver. Concomitantly, these mice showed an enhanced apoptosis in hepatocytes as Tg mice after LPS/GalN administration^[3]. Further activation of AMPK was detected in Wt mice after treatment with oligomycin A. Collectively, these data demonstrated that UCP2 expression in hepatocytes could alter mitochondrial parameters leading to activation of AMPK^[3].

References:
[1]. Jastroch M, Divakaruni AS, Mookerjee S, et al. Mitochondrial proton and electron leaks. Essays Biochemistry, 2010, 47:53-67.
[2]. Cho, J.H., M. Balasubramanyam, G. Chernaya, J.P. Gardner, A. Aviv, J.P. Reeves, P.G. Dargis, and E.P. Christian. Oligomycin inhibits storeoperated channels by a mechanism independent of its effects on mitochondrial ATP. Biochem. J. 1997.324:971-980.
[3]. Shang Y, Liu Y, Du L, Wang Y, et al. Targeted expression of uncoupling protein 2 to mouse liver increases the susceptibility to lipopolysaccharide/galactosamine-induced acute liver injury. Hepatology 2009, 50: 1204-1216