A growing body of evidence indicates a functional dichotomy between the dorsal (dHC) and ventral poles of the hippocampus (vHC). Specifically, the dorsal region is implicated in encoding spatial information and episodic memories, whereas the ventral region is recognized for its role in processing the emotional aspects associated with those memories. The medial septum/diagonal band of Broca complex (MSDB) plays pivotal role in both cognitive and emotional functions, implying anatomically distinct pathways of MSDB efferents conducting information to dHC and vHC, respectively. To test this, we traced retrogradely the MSDB neurons from both poles of the hippocampi of wild type mice. Overall, we observed a greater abundance of neurons projecting to vHC compared to those projecting to dHC. Furthermore, while similar number of cells were labeled ipsi- and contralaterally in the dHC injected brain, vHC projecting neurons of the MSDB showed lateralization, preferentially projecting to the ipsilateral HC. Using immunohistochemistry, we observed that the glutamatergic subpopulation of MSDB neurons display the most pronounced projection bias towards vHC. By employing anterograde tracing in VGluT2-cre mice, we uncovered a gradient of axonal density across all subregions along the dorso-ventral hippocampal axis. The highest density was observed in the ventral subiculum. Subsequently, using patch clamp recordings, we examined glutamatergic neurons in the MSDB, identifying heterogeneous subpopulations with overlapping firing properties that project to dHC and vHC. In summary, our findings enhance our understanding of the neural mechanisms that may underlie the distinctions in information processing between dHC and vHC.