Mitochondrial inhibition prior to oxygen-withdrawal facilitates the occurrence of hypoxia-induced spreading depression in rat hippocampal slices.

Abstract

Oxygen withdrawal blocks mitochondrial respiration. In rat hippocampal slices, this triggers a massive depolarization of CA1 neurons and a negative shift of the extracellular DC potential, the characteristic sign of hypoxia-induced spreading depression (HSD). To unveil the contribution of mitochondria to the sensing of hypoxia and the ignition of HSD, we modified mitochondrial function. Mitochondrial uncoupling by carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP, 1 microM) prior to hypoxia hastened the onset and shortened the duration of HSD. Blocking mitochondrial ATP synthesis by oligomycin (10 microg/ml) was without effect. Inhibition of mitochondrial respiration by rotenone (20 microM), diphenyleneiodonium (25 microM), or antimycin A (20 microM) also hastened HSD onset and shortened HSD duration. 3-nitropropionic acid (1 mM) increased HSD duration. Cyanide (100 microM) hastened HSD onset and increased HSD duration. At higher concentrations, cyanide (1 mM), azide (2 mM), and FCCP (10 microM) triggered SD episodes on their own. Compared with control HSD, the spatial extent of the intrinsic optical signals of cyanide- and azide-induced SDs was more pronounced. Monitoring NADH (nicotinamide adenine dinucleotide) and FAD (flavin adenine dinucleotide) autofluorescence and mitochondrial membrane potential verified the mitochondrial targeting by the drugs used. Except 1 mM cyanide, no treatment reduced cellular ATP levels severely and no correlation was found between ATP, NADH, or FAD levels and the time to HSD onset. Therefore ATP depletion or a cytosolic reducing shift due to NADH/FADH2 accumulation cannot serve as a general explanation for the hastening of HSD onset on mitochondrial inhibition. Additional redox couples (glutathione) or events downstream of the mitochondrial depolarization need to be considered.

• Authors: Florian J Gerich, Sebastian Hepp, Irmelin Probst, Michael Müller
• Journal: Journal of neurophysiology (J Neurophysiol) Vol. 96 Issue 1 Pg. 492-504 (Jul 2006) ISSN: 0022-3077 [Print] United States
• PMID: 16611842 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Azides
• Cyanides
• Enzyme Inhibitors
• Uncoupling Agents
• NAD
• Flavin-Adenine Dinucleotide
• Adenosine Triphosphate
• Oxygen

Topics

• Adenosine Triphosphate (analysis)
• Animals
• Azides (pharmacology)
• Cortical Spreading Depression (drug effects, physiology)
• Cyanides (pharmacology)
• Enzyme Inhibitors (pharmacology)
• Female
• Flavin-Adenine Dinucleotide (analysis)
• Hippocampus (chemistry, physiology)
• Hypoxia, Brain (physiopathology)
• Male
• Membrane Potentials (physiology)
• Mitochondria (physiology)
• Mitochondrial Membranes (physiology)
• NAD (analysis)
• Oxidation-Reduction
• Oxygen (physiology)
• Rats
• Rats, Sprague-Dawley
• Uncoupling Agents (pharmacology)