F<sub>1</sub>F<sub>O</sub> ATPase Vesicle Preparation and Technique for Performing Patch Clamp Recordings of Submitochondrial Vesicle Membranes

Silvio Sacchetti; Kambiz N. Alavian; Emma Lazrove; Elizabeth A. Jonas

doi:10.3791/4394

F1FO ATPase Vesicle Preparation and Technique for Performing Patch Clamp Re...

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Neuroscience

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JoVE Journal Neuroscience

F₁F_O ATPase Vesicle Preparation and Technique for Performing Patch Clamp Recordings of Submitochondrial Vesicle Membranes

F1FO ATPase Vesicle Preparation and Technique for Performing Patch Clamp Recordings of Submitochondrial Vesicle Membranes

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08:21 min

May 4, 2013

DOI: 10.3791/4394-v

Silvio Sacchetti¹, Kambiz N. Alavian¹, Emma Lazrove¹, Elizabeth A. Jonas¹

¹Department of Internal Medicine,Yale University

A method to isolate submitochondrial vesicles enriched in F1FO ATP synthase complexes from rat brain is described. These vesicles allow the study of the activity of F1FO ATPase complex and its modulation using the technique of patch clamp recording.

The overall goal of this procedure is to isolate F1 F zero APAs vesicles from rat brain for performing patch clamp recordings. This is accomplished by first removing the brain from a rat and homogenizing it. The second step is to disrupt synaptic zones by applying pressure.

Next, the synaptosome are layered onto the phial gradients. The final step is to incubate with digit toin and L to obtain the final vesicles for patch clamp recording. An important function of mitochondria is a TP production by oxidative phosphorylation.

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