This effect, independent of astrocyte P2Y1R-dependent glutamate signaling, is reminiscent of the effect mediating synaptic scaling in WT mice via surface insertion of postsynaptic AMPAR subunits ( Stellwagen and Malenka, 2006). This

observation suggests that TNFα exerts multiple, possibly coordinated, regulatory actions at excitatory synapses, which apparently converge in strengthening synaptic connectivity. Intriguingly, we did not find that basal mEPSC amplitude was reduced in Tnf−/− slices compared to WT slices (see also Beattie et al., 2002, Kaneko et al., 2008 and Stellwagen and Malenka, 2006). This is probably because TNFα mediates exclusively the scaling-up of synapses, a phenomenon in which synapses adapt to increased TNFα Trichostatin A nmr levels, whereas the opposite scaling-down phenomenon might be controlled by TNFα-independent mechanisms

( Aizenman and Pratt, 2008 and Cingolani Lenvatinib solubility dmso et al., 2008). Our study introduces the concept of regulation of the astrocytic input to synapses by ambient factors like TNFα. This is particularly relevant also because we show that the cytokine displays concentration-dependent effects on astrocytic glutamate release, going from a permissive/gating action to direct stimulation. We do not know if these represent mechanistically distinct modes of action or, perhaps more probably, a gradual shift in the effects of TNFα. In the latter case, we could hypothesize that, even at gating levels, small fluctuations in TNFα concentrations, could subtly modify the astrocytic input to synapses. Physiological processes like sleep have been proposed to be regulated by local variations in TNFα levels in the brain related to the sleep-wake cycle, and sleep deregulation can be induced by injections of the cytokine (Imeri

and Opp, 2009 and Krueger, 2008). Therefore, an intriguing hypothesis is that the TNFα control of gliotransmission is involved in sleep homeostasis together with other glial pathways already identified (Fellin et al., 2009 and Halassa et al., 2009). Moreover, the levels of TNFα are subject to dramatic changes in pathological conditions when microglia releases large amounts of the cytokine. We have already shown in a cell culture model that in such a situation, TNFα strongly amplifies glutamate release from astrocytes Tryptophan synthase (Bezzi et al., 2001). We can then hypothesize that a pathology-induced switch in the TNFα levels may have an important impact on the astrocytic input to synapses, notably in the presynaptic regulation of neuronal activity in the PP-GC hippocampal synaptic circuit. This may perturb the normal control by this circuit on critical processes such as memory formation and physiological limbic system excitability. Mice homozygous for the null mutant TNFα (Tnf−/−) allele were generated and maintained on a C57BL/6J background as described in the original study ( Pasparakis et al., 1996).