Widespread Over-Expression of the X Chromosome in Sterile F1 Hybrid Mice

The X chromosome often plays a central role in hybrid male sterility between species, but it is unclear if this reflects underlying regulatory incompatibilities. Here we combine phenotypic data with genome-wide expression data to directly associate aberrant expression patterns with hybrid male sterility between two species of mice. We used a reciprocal cross in which F1 males are sterile in one direction and fertile in the other direction, allowing us to associate expression differences with sterility rather than with other hybrid phenotypes. We found evidence of extensive over-expression of the X chromosome during spermatogenesis in sterile but not in fertile F1 hybrid males. Over-expression was most pronounced in genes that are normally expressed after meiosis, consistent with an X chromosome-wide disruption of expression during the later stages of spermatogenesis. This pattern was not a simple consequence of faster evolutionary divergence on the X chromosome, because X-linked expression was highly conserved between the two species. Thus, transcriptional regulation of the X chromosome during spermatogenesis appears particularly sensitive to evolutionary divergence between species. Overall, these data provide evidence for an underlying regulatory basis to reproductive isolation in house mice and underscore the importance of transcriptional regulation of the X chromosome to the evolution of hybrid male sterility. The X chromosome plays an important role in the development of reproductive isolation between species, but the basis for this has remained unclear. One possible explanation is that sperm development is sensitive to disruption of X-linked gene regulation. In mice, evidence linking abnormal gene expression on the X chromosome with reproductive isolation has been lacking until now. Here we use experimental crosses within and between species of mice and genome-wide expression data to identify aberrant expression patterns associated with hybrid male sterility. We observed chromosome-wide over-expression of the X chromosome during spermatogenesis in sterile hybrid males and developmentally localized this breakdown to an apparent disruption of X-inactivation. Collectively, these results highlight the importance of gene regulation to the evolution of reproductive isolation and support the hypothesis that improper expression of the X chromosome during spermatogenesis is an important mechanism contributing to the rapid evolution of hybrid male sterility.