The accurate timing of biogeographic dispersal can be determined by examining the age of fossiliferous strata on either side of a physical barrier. Here we show that African mammals migrated to Iberia and European mammals migrated to North Africa at the same time before isolation of the Mediterranean Sea during the Messinian. The fossil site of Venta del Moro (Spain) exhibits western Europe’s most complete vertebrate fauna for the latest Miocene. Its uniquely cosmopolitan assemblage is evidence of faunal dispersals from Africa and Asia to Europe during the latest Miocene glaciation. A preliminary paleomagnetic study suggested an age of 5.8 Ma for this site, but our expanded magnetostratigraphy dates the site at 6.23 Ma. In addition, we recalibrated the paleomagnetic age of the Librilla site (Spain) and the North Africa site of Afoud-1 (Morocco) using the Astronomical Tuned Neogene Time Scale. Our results show a two-way African-Iberian mammal dispersal just before 6.2 Ma. These new ages indicate that an ephemeral land corridor existed between the two continents 250 k.y. before the onset of the Messinian Salinity Crisis, reflecting a tentative initial isolation of the Mediterranean Sea. This corridor developed after tectonics closed the Betic Seaway at 6.3 Ma and during the intensification of the latest Miocene glaciation at 6.26 Ma, when water circulation in the Mediterranean became very restricted.