The house mouse, Mus musculus, provides a powerful model system for understanding evolution, and is arguably the best mammalian model for studies of the genomic basis for reproductive isolation during the early stages of speciation. It includes at least three closely related subspecies with parapatric distributions: M. m. musculus found in Eastern Europe and Northern Asia, M. m. castaneus in Southeast Asia and India, and M. m. domesticus, in western Europe, the Near East, and northern Africa [1]. These three subspecies rapidly diverged in allopatry around 350,000 years ago [2][3][4], and evidence suggests that M. m. castaneus and M. m. musculus are more closely related to each other than either is to M. m. domesticus [5, 6]. During the past 10,000 years, house mice have become commensal with humans, and as stowaways with them, have become the most successful small mammal colonizers of new continents during the past few hundred years [7,8].Regions of secondary contact and introgression may mark where mouse subspecies meet in nature. The best studied of these is a narrow hybrid zone between M. m. domesticus and M. m. musculus that stretches from Denmark to the Black Sea in central Europe [9][10][11][12][13][14][15][16][17][18]. This hybrid zone is young, with mice having colonized this *Author for correspondence (andrew.j.veale@gmail.com).†Present address: Department of Zoology, University of Otago, 340 Great King St, Dunedin 9016, New Zealand . CC-BY-NC-ND 4.0 International license peer-reviewed) is the author/funder. It is made available under a The copyright holder for this preprint (which was not . http://dx.doi.org/10.1101/234245 doi: bioRxiv preprint first posted online Dec. 14, 2017; area around 3000 years ago [19, 20]. Hybridization in the wild between M. m. domesticus and M. m. castaneus is best known from one study of an introduced population in California [21] and one in New Zealand [22]. Within the native range, other possible domesticus/castaneus hybrid zones in Iran [23][24][25] and in Indonesia [26] have produced only preliminary results, because these regions are complex, supporting multiple (and potentially undescribed) subspecies [24], and because comprehensive nuclear loci have not been used to look at the levels of admixture across the genomes.Hybrids between subspecies have been extensively studied in laboratory strains of mice, with data indicating that M. m. musculus and M. m. domesticus are largely reproductively isolated [27,28]. These studies have helped us to understand the genetics of speciation, particularly the genetic basis of hybrid male sterility [29][30][31][32], revealing an important role of the X chromosome in producing reproductive incompatibilities [33][34][35][36][37][38][39][40]. Studies of both wild and laboratory mice have also found that hybrid male sterility has a complex basis, involving many genes [31,32,38,41]. Laboratory crosses between M. m. domesticus and M. m. musculus led to the identification of Prdm9 on chromosome 17, the only gene at present known to cont...