The entorhinal cortex (EC) connects to the hippocampus sending different information from cortical areas that is first processed at the dentate gyrus (DG) including spatial, limbic, and sensory and information. Excitatory afferents from lateral (LPP) and medial (MPP) perforant pathways of the EC connecting to granule cells (GC) of the DG play a role in memory encoding and information processing. LPP- and MPP-GC synapses have distinct electrophysiological properties, and these differences potentially affecting the functional processing of information. However, the plasticity of these synapses is not well known yet, as are not known the forms of long-term depression (LTD) existing at those connections. We investigated whether spike timing-dependent long-term depression (t-LTD) exists at these two different EC-DG synaptic connections in mice, and whether they have different action mechanisms. We have found two different forms of t-LTD, at LPP- and MPP-GC synapses, and characterised their cellular and intracellular mechanistic requirement. The most relevant results showed that both forms of t-LTD are expressed presynaptically and that whereas t-LTD at MPP-GC requires ionotropic NMDAR containing GluN2A subunits, t-LTD at LPP-GC synapses does not require NMDAR. In addtion, the two forms of t-LTD require different group I mGluR, postsynaptic calcium, synthesis and release of endocannabinoids by the postsynaptic cell, as well as astrocyte activity releasing glutamate. Therefore, we discovered two novel forms of presynaptic t-LTD that requires astrocytes at EC-GC synapses with different action mechanisms, which could contribute to the functional processing of the different information.