Selective protection by phosphatidic acid against staurosporine‐induced neuronal apoptosis

Abstract

Phosphatidic acid, the main product of lipid breakdown through phospholipase D activation, has been implicated in important signal transduction pathways able to influence cell fate in many ways. The purpose of this work was to determine possible effects of phosphatidic acid on neuronal cell death pathways. Here we used cerebellar granular cell cultures and cell death was triggered with either staurosporine or H2O2. Cell viability was quantified by spectrophotometry, using the 3‐ (4, 5‐dimethylthiazol‐2‐y1)‐2, 5‐diphenyl‐tetrazolium bromide (MTT) test. Staurosporine (1‐3 μM) or H2O2 (50‐800 μM) induced cell death in a dose‐dependent manner. Using fluorescent staining (propidium iodide or annexin V‐Cy3/6‐carboxyfluorescein) we showed that cell death was mostly apoptotic in staurosporine treated cells and mostly non‐apoptotic (necrotic) in H2O2 treated cells. Phosphatidic acid was able to increase cell viability in staurosporine‐, but not in H2O2 ‐ treated cells. We therefore conclude that phosphatidic acid has neuroprotective potential in neurons exposed to stimuli that trigger apoptosis.

Expression of phospholipase D isozymes in scar and viable tissue in congestive heart failure due to myocardial infarction

“…Although an oleate-dependent PLD activity is drastically increased during apoptosis of Jurkat T cells [38], an increased PLD2 activity has been shown to reduce hypoxiainduced death of PC12 cells [39], while a hypoxia-induced activation of PLD1 in smooth muscle cells has also been identified [40]. Furthermore, PA has been reported to be protective against staurosporine-induced apoptosis in cultured neuronal cells [41]; these studies suggest that PLD isozymes play a role in cellular apoptosis. It is interesting to note that in smooth muscle cells, activation of the phosphatidylinositol-specific phospholipase C (PLC) is essential for triggering contraction, while PLA 2 is involved in the sustained tonic contraction and PLD appears to play a role in triggering relaxation [42,43].…”

Dantrolene protects neurons against kainic acid induced apoptosis in vitro and in vivo

“…Primary CGC cultures were prepared as previously described [26]. Cell culture supplies were purchased from Nunc A/S (Roskilde, Denmark) and foetal calf serum, Dulbecco's modified Eagle's medium (DMEM), L-glutamine, penicillin-streptomycin, phosphate buffered saline (PBS) and trypsin were from Sigma (St. Louis, MO, USA).…”

Gabapentin is neuroprotective through glutamate receptor-independent mechanisms in staurosporine-induced apoptosis of cultured rat cerebellar neurons

“…hours in a previous study [27]. In this study we Even though it is well-documented that STS triggers apoptosis independent of glutamate and GABA receptors [34], considering that endogenous glutamate is released in CGN cultures and that these cultures are sensitive to changes in receptor-operated intracellular ionic concentrations [31] and that the main mechanism of TPM neuroprotection is the blockade of AMPA receptors [24], we tested whether we can reverse the protective effect of GP and TPM by GABA or by glutamate ionotropic receptor antagonists.…”

“…on a previous study [27] in order to have a partial and potentially reversible induction of apoptosis in CGN cultures.…”

“…We also tested effects of GP and TPM on the expression of neuronal cell adhesion molecule (NCAM) and synaptophysin (SP), proteins which are important for neuronal plasticity [25,26]. Primary rat CGN cultures were prepared as previously described [27] and grown in all The control cells were exposed to vehicle (DMSO) in growth medium for 6 hours. At least 3 different experiments were run for each experimental situation.…”