Back to Search
Start Over
Comparative evaluation of hypoxic-ischemic brain injury by flow cytometric analysis of mitochondrial membrane potential with JC-1 in neonatal rats
- Source :
- Journal of neuroscience methods. 193(2)
- Publication Year :
- 2010
-
Abstract
- We assessed the validity of monitoring changes in mitochondrial membrane potential (ΔΨ) with a fluorescent probe, JC-1 (5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl benzimidazolo-carbocyanine iodide), for the quantitative evaluation of neonatal hypoxic-ischemic brain injury. Seven-day-old rat pups were subjected to 2h of 8% oxygen following unilateral carotid artery ligation. Brain tissue was obtained for JC-1 staining at 24h after hypoxia ischemia (HI), and the results were compared with those of other simultaneous measurements such as flow cytometry with fluoresceinated annexin V/propidium iodide (PI), terminal deoxynucleotidyltransferase-mediated dUTP nick end-labeling (TUNEL) staining, triphenyl tetrazolium chloride (TTC) infarct area and western blot for cytosolic cytochrome c. Flow cytograms of JC-1 showed two distinct sub-populations with different ΔΨ, red with high ΔΨ and green with low ΔΨ, at 24h after HI. This shift of JC-1 fluorescence from red to green indicated a collapse of ΔΨ. The increased percentage of low ΔΨ with JC-1 showed a significant positive correlation with a simultaneous increase in annexin V(+)/PI(+) necrotic cells, TUNEL-positive cells, TTC infarct area and western blot of cytosolic cytochrome c, and negative correlation with annexin V(-)/PI(-) live cells. In summary, low ΔΨ measured with JC-1 was significantly correlated with results from other methods used to assess the extent of brain damage after HI. Therefore, fluorocytometric analysis of ΔΨ with JC-1 might be a sensitive and reliable technique in the quantitative evaluation of neonatal brain injury.
- Subjects :
- Brain Infarction
Time Factors
Tetrazolium Salts
Brain damage
Biology
Flow cytometry
Rats, Sprague-Dawley
chemistry.chemical_compound
Western blot
Annexin
medicine
In Situ Nick-End Labeling
Animals
Propidium iodide
Annexin A5
Fluorescent Dyes
Membrane Potential, Mitochondrial
Neurons
TUNEL assay
medicine.diagnostic_test
Cell Death
Tetrazolium chloride
General Neuroscience
Brain
Cytochromes c
Carbocyanines
Flow Cytometry
Molecular biology
Staining
Rats
Disease Models, Animal
chemistry
Animals, Newborn
Hypoxia-Ischemia, Brain
Benzimidazoles
medicine.symptom
Propidium
Subjects
Details
- ISSN :
- 1872678X
- Volume :
- 193
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Journal of neuroscience methods
- Accession number :
- edsair.doi.dedup.....09332552da80e62a4a3a8663aa0bf3e8