The development of two-photon imaging techniques has created new ways to study the function of the mouse brain. We are using this method to monitor in vivo the activation of a FosGFP reporter in the retrosplenial cortex (RSC) in mice with chronically implanted optical windows. This study was designed to explore the mechanisms of RSC involvement in spatial memory encoding. RSC receives projections from the anterior thalamus and the CA1 region of hippocampus; it is also reciprocally connected with the subiculum, anterior cingulate and parietal cortices. Therefore this structure has been proposed to have a critical role in the circuitry responsible for spatial learning and memory. For the imaging experiment mice were trained for 10 days alternatively in a “spatial” or in a “cued” version of the Morris water maze task (MWM). In the “spatial” version (days 1, 3, 5, 7 and 9 of training) mice could use extra-maze cues in the experimental room. In the “cued” version (days 2, 4, 6, 8 and 10), the extra-maze cues were blocked with a curtain placed around the pool and the platform position was marked with metal rod. On days 7–10, 90 minutes after the 4th and 5th sessions, specifi c regions of the RSC were imaged in lightly anesthetized FosGFP mice. Analysis revealed that approximately 30% of cells imaged showed reliable changes in activity when alternating between the spatial and cued task. Among those cells, the majority showed higher activity after the spatial task than after the cued task.
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