Inhibition of Mitochondrial Permeability Transition Pore Evaluated by Calcium Retention Capacity Assay

The pathogenesis of ischemia-reperfusion injury of heart, acute pancreatitis, neurodegeneration and brain and kidney, muscular dystrophies, have a common key characteristic - mitochondrial abnormality.

A drastic rise in permeability of inner mitochondrial membrane (IMM), through consistent opening of a multi-protein channel referred to as the mitochondrial permeability transition pore (MPTP) results in mitochondrial dysfunction.

Following this, uncontrolled proton flow through the IMM occurs along with unregulated transfer of ions, water, and small solutes across the mitochondrial matrix. Rupture of outer mitochondrial membrane (OMM) and ultimately cell death due to necrosis are the end results. Hence, for the prevention of cell death in a variety of disease conditions, the MPTP is a potential target.

The calcium retention capacity technique uses spikes of calcium ions to challenge the isolated mitochondria. After opening of MPTP, fluorescence of membrane-impermeable CalciumGreen™ dye is enhanced as Ca2+ leaches into the buffer. The Omega multi-mode plate reader has proven outstanding robustness for carrying out multiple injections. Thedevice has also shown a reliable fluorescent recognition of the assay.

Read the full article including the methods, results and discussion.

Calcium retention capacity assay evaluates inhibition of mitochondrial permeability transition pore

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Last updated: Mar 28, 2019 at 5:38 AM

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