MEMRI can be used for different applications such as tracing neuronal connections or functional imaging. However, Mn2+ uptake and transport mechanisms are still unclear. These mechanisms were studied in an organotypic hippocampal slice culture. After adding Mn to the medium, MRI contrast changes were affected by Ca channel manipulation and other metals that use divalent metal transporters. Mn cellular uptake was also affected by both, the presence of glutamate receptor antagonist and a neuronal activity inhibitor. Our results establish hippocampal slice as a system to study the cellular mechanisms related to MEMRI.