Alzheimer Disease’s (AD) etiology is a hot topic in neuroscience. Cholinergic hypothesis was the earliest theory while amyloidogenic hypothesis is currently the most studied. Unfortunately, despite massive and prolonged efforts, triggering events responsible for AD genesis remain elusive. Here, we aim to elucidate the cellular and molecular mechanisms that induce AD by focusing on glutamatergic tripartite synapses in hippocampus. It is known that Aβ binds to α7 Acetylcholine nicotinic receptors (α7nAchRs) with high affinity, favoring synaptic plasticity at physiological picomolar concentrations. We thus take advantage of an age-related AD-like mouse model in which functionally knocking-out α7nAchRs leads to Aβ accumulation, hyperphosphorylated Tau and cognitive impairments similarly to AD (Tropea et al 2021). We use 2-photon laser scanning microscopy to monitor glutamate release in hippocampal brain slices expressing SF-iGluSNFr sensor. Results show different glutamate release in young compared to older WT and α7KO mice. In addition, response to acute administration of Aβ changes in relation to age and phenotype. In slices from young WT mice, glutamate release is increased by Aβ whereas in slices from α7KO mice it is reduced. The lack of functional α7nAchRs may induce Aβ overproduction, as previously suggested, that in the long term may alter glutamate synapses with different mechanisms. The role of astrocytic glutamate uptake is also explored. Further investigation will clarify the specific role of astrocytic and neuronal a7nAchRs. Our data may help clarify how Aβ switches from physiological to pathological by acting on cholinergic and glutamatergic systems in AD onset and progression.