Beta-adrenergic receptors (B-ARs) regulate hippocampal synaptic plasticity associated with spatial memory in rodents (doi: 10.1002/hipo.22168). Synaptic plasticity supports the refinement and stabilisation of neuronal ensembles. This study aimed to examine the effect of noradrenergic regulation of dorsal hippocampal neuronal activity during spatial learning. For this, we used wide-field calcium imaging as a method to study B-AR modulation of the activity of visualised hippocampal CA1 neurons during item-place learning that is known to trigger hippocampal long-term depression (LTD) (doi: 10.1093/cercor/bhs089). Our results revealed that pharmacological antagonism of B-AR prevented the acquisition of item-place memory in mice. Imaging of neuronal activity in the CA1 region during the acquisition, retrieval and updating of item-place information revealed that the neuronal population recruited in control mice during de novo spatial learning was stabilized during retrieval and further refined during information updating. Thus, control animals showed a reduction in overall recruitment activity during the course of item-place learning. This suggests that processing of item-place associations was achieved through ensemble optimization. In contrast, B-AR antagonism led to an elevated level of neuronal recruitment during acquisition, retrieval and information updating; indicating that in the absence of B-AR activation ensemble optimization fails to occur. In conclusion, these results indicate that B-ARs play a crucial role in establishing robust spatial representations by modulating network-level stability and refinement which supports encoding and retrieval of spatial information by the hippocampus.Supported by the Deutsche Forschungsgemeinschaft (SFB 1280/A04, project number: 316803389).