Involvement of inositol 1,4,5-trisphosphate-regulated stores of intracellular calcium in calcium dysregulation and neuron cell death caused by HIV-1 protein Tat

HIV-1 infection commonly leads to neuronal cell death and a debilitating sy ndrome known as AIDS-related dementia complex. The HIV-1 protein Tat is neu rotoxic, and because cell survival is affected by the intracellular calcium concentration ([Ca2+](i)), we determined mechanisms by which Tat increased [Ca2+](i) and the involvement of these mechanisms in Tat-induced neurotoxi city. Tat increased [Ca2+](i) dose-dependently in cultured human fetal neur ons and astrocytes, In neurons, but not astrocytes, we observed biphasic in creases of [Ca2+](i). Initial transient increases were larger in astrocytes than in neurons and in both cell types were significantly attenuated by an tagonists of inositol 1,4,5-trisphosphate (IP3)-mediated intracellular calc ium release [8-(diethylamino)octyl-3,4,5-trimethoxybenzoate HCI (TMB-8) and xestospongin], an inhibitor of receptor-G(i) protein coupling (pertussis t oxin), and a phospholipase C inhibitor (neomycin). Tat significantly increa sed levels of IP3 threefold. Secondary increases of neuronal [Ca2+](i) in n eurons were delayed and progressive as a result of excessive calcium influx and were inhibited by the glutamate receptor antagonists ketamine, MK-801, (+/-)-2-amino-5-phosphonopentanoic acid, and 6,7-dinitroquinoxaline-2,3-di one. Secondary increases of [Ca2+](i) did not occur when initial increases of [Ca2+](i) were prevented with TMB-8, xestospongin, pertussis toxin, or n eomycin, and these inhibitors as well as thapsigargin inhibited Tat-induced neurotoxicity. These results suggest that Tat, via pertussis toxin-sensiti ve phospholipase C activity, induces calcium release from IP3-sensitive int racellular stores, which leads to glutamate receptor-mediated calcium influ x, dysregulation of [Ca2+](i), and Tat-induced neurotoxicity.