Phospholipase C is involved in the adenosine-activated signal transduction pathway conferring protection against iodoacetic acid-induced injury in primary rat neuronal cultures

We have demonstrated before that exposure of neuronal cultures to poisoning by iodoacetic acid, followed by "reperfusion" (iodoacetate- "reperfusion" insult; IAA-R insult), results in severe cytotoxicity. This insult was found to be associated with ATP depletion and generation of reactive oxygen species. The cultured neurons could be protected against the insult by activation of the adenosine A₁ receptors and by presence of antioxidants. Previous studies in our laboratory demonstrated that the adenosine-activated signal transduction pathway (Ado-STP) conferring protection against the IAA-R insult, involves activation of protein kinase C-epsilon (PKCε) and opening of ATP sensitive potassium (K_ATP) channels. In this respect, the adenosine-activated protective mechanism against the IAA-R insult is similar to the Ado-STP in the neurons and in cardiomyocytes against ischemia-reperfusion injury. Phospholipase C (PLC) is an additional component demonstrated recently to participate in the myocardial Ado-STP protecting against ischemia-reperfusion. Here we provide proof for the involvement of PLC also in the neuronal Ado-STP protecting against the IAA-R insult. Primary rat neuronal cultures were exposed to the IAA-R insult. The neurons could be protected against this insult by activation of the adenosine A₁ receptors by N⁶-(R)-phenylisopropyladenosine (R-PIA), a specific A₁ adenosine receptor agonist. Exposure of the cultures to the PLC inhibitor U73122, abrogated the protection. The exposure of the cultures to R-PIA was found to enhance PLC activity, an effect that could be abrogated by presence of U73122. The R-PIA-induced increase in PLC activity was short-lived, in the range of minutes. These results demonstrate that activation of PLC is a vital step in the neuronal protective Ado-STP, but that it does not contribute directly to the relatively long time window of the protection signal shown previously to characterize the neuronal mechanism. The results also support the suggestion that the Ado-STP protecting against the IAA-R insult and that protecting against ischemia-reperfusion may represent the same mechanism.