The hippocampus and entorhinal cortex play a pivotal role in spatial learning and memory. The two forebrain regions are highly interconnected via excitatory pathways. Using optogenetic tools, we identified and characterized long-range γ-aminobutyric acid-releasing (GABAergic) neurons that provide a bidirectional hippocampal-entorhinal inhibitory connectivity and preferentially target GABAergic interneurons. Activation of long-range GABAergic axons enhances sub- and suprathreshold rhythmic theta activity of postsynaptic neurons in the target areas.

SummaryLayer II (LII) of the medial entorhinal cortex (MEC) comprises grid cells that support spatial navigation. The firing pattern of grid cells might be explained by attractor dynamics in a network, which requires either direct excitatory connectivity between phase-specific grid cells or indirect coupling via interneurons. However, knowledge regarding local networks that support in vivo activity is incomplete. Here we identified essential components of LII networks in the MEC. We distinguished four types of excitatory neurons that exhibit cell-type-specific local excitatory and inhibitory connectivity. Furthermore, we found that LII neurons contribute to the excitation of contralateral neurons in the corresponding layer. Finally, we demonstrated that the medial septum controls excitation in the MEC via two subpopulations of long-range GABAergic neurons that target distinct interneurons in LII, thereby disinhibiting local circuits. We thus identified local connections that could support attractor dynamics and external inputs that likely govern excitation in LII.