The Japanese dormouse, Glirulus japonicus, is the only extant lineage that represents this genus and it has been classified as a single species distributed on the three main islands of Japan, namely Honshu, Shikoku and Kyushu. However, individuals collected from Fukui, Wakayama and Kochi Prefectures (southwestern part of Japan) yielded distinctly different profiles of restriction fragments of the nuclear ribosomal DNA (rDNA) spacer from those collected from Yamanashi and Nagano Prefectures (central Japan). The estimated sequence divergence between the two groups was 2.8% on average, which corresponds to a putative divergence some two million years ago. Representing mitochondrial DNA (mtDNA) sequences, 402 bases of cytochrome b gene were determined by direct sequencing and the estimated extent of the sequence divergence between the two groups was 6.5–7%. Differences between the two geographic groups were also substantial in the sequences of about 300 base-fragments from the Y-linked, sex-determining locus, Sry. To assess the phylogenetic relationships between the Japanese dormouse and members of the family Myoxidae, we compared sequences of mitochondrial 12S rRNA gene of Japanese dormice with those of the forest dormouse (Dryomys nitedula) and the common dormouse (Muscardinus avellanarius), two continental genera thought to be closely related to the genus Glirulus. The results showed that the sequences from Japanese dormice were distinct from any sequences of the two continental species and the extent of the differences were somewhat similar to that between the rat (Rattus norvegicus) and the hamster (Mesocricetus auratus).