The eastern border of the Middle Valley of the Tiber River is characterized by several Plio-Pleistocene paleoshorelines, which extend for about 100 km along the western margin of the Central Apennines (Italy). We studied these paleoshorelines by the means of geological and paleontological analyses and new 87Sr/86Sr isotope analyses. The youngest and uppermost paleoshorelines have been detected and mapped through detailed geologic and stratigraphic surveys, which led to the recognition of nearshore deposits, cliff breccias, alignments of Lithophaga borings, fossil abrasion notches and wave-cut platforms. The altitude of these paleoshorelines decreases almost regularly in the NNW–SSE direction from 480 to 220 m a.s.l. Measurements of the 87Sr/86Sr isotope ratio have been conducted on corals and mollusks collected from sediments outcropping close to the paleoshorelines. The isotopic dating results indicate numerical values that range between 0.70907 and 0.70910 all over the 100-km outcrop. These results, together with biostratigraphic data, constrain the age of the youngest paleoshorelines to 1.65–1.50 Ma. These paleoshorelines are thus considered almost isochronous, giving an estimated uplift rate of 0.34–0.17±0.03 mm/a moving from NNW to SSE. Shape, length and continuity of the 100-km-long observed movements indicate that the studied paleoshorelines are an important marker of the Quaternary uplift of the Central Apennines.