Evidence against calcium as a mediator of mitochondrial dysfunction during apoptosis induced by arachidonic acid and other free fatty acids.
Apoptosis is often accompanied by activation of phospholipase A(2), causing release of free fatty acids (FFAs), which in turn are thought to contribute to the loss of mitochondrial transmembrane potential (Deltapsi(m)). In these experiments, we asked whether calcium plays a role as an intermediate in this process. A total of 14 FFAs were compared for their ability to cause loss of Deltapsi(m) and for their ability to affect levels of intracellular calcium. Among the FFAs, unsaturated FFAs tended to induce apoptosis while saturated FFAs did not. Arachidonic acid (AA) was most damaging, causing loss of Deltapsi(m) and cell death in 8-10 h while linoleic acid, gamma-linolenic acid, and docosapentaenoic also strongly induced apoptosis. Effects of the FFAs on levels of intracellular calcium were very different. Many caused strong calcium responses; however, the ability to induce a strong calcium response was not predictive of ability to induce apoptosis, and overall, we did not find a correlation between apoptosis and calcium induction. Also, verapamil and TMB-8 were able to block the calcium response, but these inhibitors did not prevent loss of Deltapsi(m), indicating that the calcium response is not necessary for FFA-induced loss of Deltapsi(m). In contrast, we found that cyclosporine A could inhibit the AA-induced loss of Deltapsi(m) with both whole cells and isolated mitochondria, confirming that the antimitochondrial effects of FFA can stem from direct effects on the mitochondrial permeability transition pore. Finally, we show that the strong apoptosis-inducing activity of AA may stem from its ability to selectively induce its own release.
Maia
Rita C
RC
Department of Microbiology, North Carolina State University, Raleigh, NC 27695, USA.
Culver
Carolyn A
CA
Laster
Scott M
SM
D000818:Animals; D017209:Apoptosis; D016718:Arachidonic Acid / Q000494:pharmacology*; D019898:Autocrine Communication; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D005230:Fatty Acids, Nonesterified / Q000494:pharmacology*; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D051379:Mice; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism
D005230:Fatty Acids, Nonesterified; D016718:Arachidonic Acid; D002118:Calcium