The Eastern Arc Mountains of eastern Africa are notable for the high levels of endemism exhibited by various forest-dwelling organisms of this ancient montane archipelago. There has been virtually no assessment of the variation among populations of small mammal species living on these unique mountains, but recent faunal surveys have produced sufficient material to initiate such studies. Cranial morphometric and DNA sequence data were examined from six populations of Sylvisorex howelli Jenkins, 1984, an endemic shrew found in several different massifs of the Eastern Arc Mountains, to assess variation across the archipelago in the context of various hypotheses of historical biogeography. Twenty-two cranial measurements were analysed using principal components analysis. Age classes (based on tooth wear) and sex had little effect on the variation exhibited by the variables studied. Overall, specimens of S. howelli from the East Usambara Mountains are smaller than specimens from other known populations. The mitochondrial ND2 and 12S rRNA genes from representatives of each montane population of S. howelli in addition to several crocidurine taxa from eastern Africa and three soricine outgroup species were sequenced to assess phylogenetic relationships among these taxa. Neither maximum likelihood, maximum parsimony, nor Bayesian analyses support monophyly of the genus Sylvisorex, but S. howelli populations were consistently recovered as a well-supported clade. Over 40 individuals of S. howelli from six disjunct montane ranges, comprising the entire known distribution of the species, were sequenced for 504 base pairs of ND2 to investigate phylogeographic patterns. Phylogenetic analysis recovered six reciprocally monophyletic haplotype clades grouped by locality. Branch lengths are consistent with relatively long periods of isolation among populations from the Uluguru, Ukaguru, Nguru, Nguu, East Usambara and West Usambara Mountains, with low levels of diversity observed within each population. These results are interpreted within the historical context of the Eastern Arc Mountains.