Rapid Determination of the Membrane Potential of Mitochondria in Small Biopsy Specimens of the Liver

Abstract

In liver transplantation, graft viability is ideally to be determined before implantation. Integrity of mitochondria may be a prerequisite to a viable graft. A new method is presented, which allows for the determination of the membrane potential of mitochondria (MPM; mV) in state 4 respiration within 50 min in 40-mg specimens, employing rhodamine 123 as a probe. Normal control showed a MPM of 239.2 mV. Storage in saline at 37°C yielded an impaired MPM of 153.5 mV within 3 h. The cold storage at 1 °C could preserve MPM at quasi-normal after 3 h but reduced it significantly after 24 h to 222.2 mV in saline (p < 0.005 vs. control) and 231.0 mV in UW solution (p < 0.05 vs. control): the difference between the 24-hour values was significant (p < 0.05).

Inhibition of tumor necrosis factor‐induced apoptosis in transgenic mouse liver expressing creatine kinase

“…The final pellet was suspended in 4 ml medium (pH=7.4) consisting of 0.28 M mannitol, 0.01 M Tris buffer, 0.01 M KCl, 0.2 mM EDTA Tris, 3 mM phosphate buffer, 1.2 mM MgCl 2 , and 3 mM l‐glutamate. Ψ m was measured as that of mitochondria in state 4 respiration in the presence of glutamate without exogenous ADP using the method of Yamamoto et al, previously described (17, 18), with rhodamine 123. In brief, 3 ml mitochondria suspension and 3 ml medium containing 2 μg/ml rhodamine 123 were incubated at 30°C for 5 min.…”

Heat Shock Preconditioning on Mitochondria during Warm Ischemia in Rat Livers

“…Membrane permeability of the mitochondria was assessed by the changes in mitochondrial membrane potential (MPM: mV) as they are consuming glutamate in the absence of adenosine diphosphate (ADP). At state 4 respiration, MPM was measured using rhodamine-123 as a probe according to the method of Yamamoto et al [30]. Respiratory functions of mitochondria were measured polarographically with a rotating electrode at 22°C in a reaction medium containing 0.3 M mannitol, 10 mM KCl, 40 mM Tris-HCl, 0.2 mM EDTA-Tris, 2 mM MgCl 2 , and 3 mM phosphate buffer at a pH of 7.4.…”

Characterization of Spermatozoa of the Lizard Cnemidophorus sexlineatus Using Fluorescence Microscopy

“…accumulates in membrane-bound cellular compartments in response to the potential difference across the plasma membrane. Thus, in living cells Mitotracker preferentially accumulates in active mitochondria (e.g., Yamamoto et al, 1995). In this investigation, a concentration of 0.17 pg/p1 of this dye was selected.…”