The Expression Profile of the Major Mouse SPO11 Isoforms Indicates that SPO11β Introduces Double Strand Breaks and Suggests that SPO11α Has an Additional Role in Prophase in both Spermatocytes and Oocytes

Abstract

Both in mice and humans, two major SPO11 isoforms are generated by alternative splicing: SPO11α (exon 2 skipped) and SPO11β. Thus, the alternative splicing event must have emerged before the mouse and human lineages diverged and was maintained during 90 million years of evolution, arguing for an essential role for both isoforms. Here we demonstrate that developmental regulation of alternative splicing at the Spo11 locus governs the sequential expression of SPO11 isoforms in male meiotic prophase. Protein quantification in juvenile mice and in prophase mutants indicates that early spermatocytes synthesize primarily SPO11β. Estimation of the number of SPO11 dimers (ββ/αβ/αα) in mutants in which spermatocytes undergo a normal number of double strand breaks but arrest in midprophase due to inefficient repair argues for a role for SPO11β-containing dimers in introducing the breaks in leptonema. Expression kinetics in males suggested a role for SPO11α in pachytene/diplotene spermatocytes. Nevertheless, we found that both alternative transcripts can be detected in oocytes throughout prophase I, arguing against a male-specific function for this isoform. Altogether, our data support a role for SPO11α in mid- to late prophase, presumably acting as a topoisomerase, that would be conserved in male and female meiocytes.

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Supplemental material for this article may be found at http://mcb.asm.org/.

The Intramural Research Program of the NIDDK, NIH, supported this research.

We thank Bruce Raaka for his help with the FACS of testicular cells, Eric Anderson for his help with the mass spectrometry analysis, and Huiyan Lu for her help with oocyte isolation. We are grateful to Michael Lichten for a critical reading of our manuscript. Our thanks go to John Schimenti for providing us with Dmc1^−/− mice, to Florencia Pratto for countless insightful discussions, and to Linda Robinson for her assistance.