JoVE Encyclopedia of Experiments
Neuroscience
0 views • 4:02 min • April 28th, 2025
Place brain slice punches from healthy and neurodegenerative mice on sample-holding screens.
Flip the screens and insert them into a tissue plate.
Take a pre-calibrated sensor cartridge with sensor probes. Load the compounds—pyruvate, oligomycin, FCCP, and antimycin A—into the injection ports.
Place the cartridge on the tissue plate. Initiate the run in an extracellular flux analyzer.
The probes measure the slice’s oxygen consumption rate or (OCR).
With the first injection, pyruvate enters the mitochondria, generating electron carriers.
These carriers donate electrons to Complexes I and II of the electron transport chain or (ETC), initiating an electron transfer reaction that increases oxygen consumption and OCR.
Oligomycin inhibits Complex V, halting ATP synthesis and decreasing OCR.
FCCP, a protonophore, disrupts the proton gradient, increasing oxygen consumption and OCR.
Antimycin A binds to Complex III and stops the ETC, minimizing OCR.
Compare the OCR profiles.
A decreased OCR in the neurodegenerative slices indicates mitochondrial impairment.
Suction one piece of punched brain tissue, and carefully place the tissue onto the mesh side of the capture screen. A circular piece of plastic w
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