Cajal-Retzius neurons (CRs) are a transient cell type that populates the postnatal hippocampus. To test how the persistence of CRs shapes the maturation of hippocampal function, we used a CRs-specific transgenic mouse line combined with targeted viral delivery to selectively ablate CRs in the postnatal hippocampus. Single cell sequencing revealed that gene networks in superficial CA1 pyramidal cells were more strongly perturbed compared to deep CA1 pyramidal cells. To test if these two subpopulations were also distinctly affected in their function, we performed in vivo recordings from spatially modulated cells in CA1. Our analysis showed an impaired spatial representation specifically in superficial CA1 pyramidal cells. Additionally, we observed an increased CA3 to CA1 excitatory drive, as indicated by increased gamma oscillations, and alterations of intrinsic firing properties in superficial CA1 pyramidal neurons confirmed by in vitro electrophysiological recordings. Together, these results indicate a crucial role for CRs in the maturation of hippocampal sub-circuits.