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The present study aims to depict the overall pattern of Acomys history in south-western Sahara. We tested the specific predictions that: (1) several mitochondrial clades can be identified and that they coincide with the described species; (2) successive phases of desert expansion and contraction during the last 3 Myr have resulted in several phases of demographic expansion and population fragmentation in Acomys; and (3) isolation-by-distance occurs. The extent of phylogeographic patterns and molecular genetic diversity (cytochrome b gene and D-loop) were addressed in a survey of 90 individuals of Acomys from 38 localities. Our phylogeographical analyses show a strong genetic structure within western Saharan Acomys, with several phylogroups displaying non-overlapping geographic distributions. Restricted gene flow with isolation-by-distance was recorded and a signal of population expansion was detected within several clades. We suggest that during arid or semi-arid paleoclimatic periods, when large sandy areas were present, Acomys was restricted to rocky massifs, whereas, in more humid periods, when savannah and/or steppe habitat prevailed, this species was able to disperse and to survive outside rocky areas because food resources were abundant. Based on a comprehensive sampling and the use of an integrative approach (i.e. combining cytogenetic, molecular and morphological data), we firmly propose that Acomys airensis should be considered as a junior synonym of Acomys chudeaui. (C) 2009 The Linnean Society of London, Biological Journal of the Linnean Society, 2009, 98, 29-46.