Alterations in Membrane Potential in Mitochondria Isolated from Brain Subregions During Focal Cerebral Ischemia and Early Reperfusion: Evaluation Using Flow Cytometry

Mitochondria isolated from brain tissue following middle cerebral artery occlusion or during early reperfusion were tested for their ability to generate a membrane potential under standard conditions in vitro. Membrane potential was evaluated based on rhodamine 123 fluorescence in the mitochondria a...

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Bibliographic Details
Published in:Neurochemical research 2009-10, Vol.34 (10), p.1857-1866
Main Authors: Lee, Diane R., Helps, Stephen C., Macardle, Peter J., Nilsson, Michael, Sims, Neil R.
Format: Article
Language:English
Subjects:
Animals
Biochemistry
Biomedical and Life Sciences
Biomedicine
Brain - blood supply
Brain - pathology
Brain - physiology
Brain Ischemia - pathology
Brain Ischemia - physiopathology
Cell Biology
Cell Separation - methods
Flow cytometry
Flow Cytometry - methods
Focal ischemia
Intracellular Membranes - pathology
Intracellular Membranes - physiology
Male
MEDICAL AND HEALTH SCIENCES
MEDICIN OCH HÄLSOVETENSKAP
Membrane Potentials - physiology
Mitochondria
Mitochondria - physiology
Mitochondrial membrane potential
Neurochemistry
Neurology
Neurosciences
Original Paper
Rats
Rats, Sprague-Dawley
Reperfusion
Reperfusion Injury - pathology
Reperfusion Injury - physiopathology
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Summary:Mitochondria isolated from brain tissue following middle cerebral artery occlusion or during early reperfusion were tested for their ability to generate a membrane potential under standard conditions in vitro. Membrane potential was evaluated based on rhodamine 123 fluorescence in the mitochondria as detected using flow cytometry. Compared with equivalent samples from the contralateral hemisphere, the geometric mean fluorescence was significantly lower in mitochondria prepared from the striatum and perifocal tissue in the cortex at 3 h ischemia. During reperfusion, this property was decreased in mitochondria from tissue in the striatum and cortex that had been part of severely ischemic core tissue during the arterial occlusion. These findings provide additional evidence that mitochondria develop changes during ischemia and reperfusion that are likely to limit their ability to respond to changing energy requirements and contribute to cell dysfunction and cell death. It also demonstrates the ability to gain a sensitive measure of these mitochondrial changes using flow cytometry.
ISSN:0364-3190
1573-6903
1573-6903
DOI:10.1007/s11064-009-0001-1