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sperm Motility Assay
Sperm were diluted at least 1:10 in MVF. The samples were then drawn into
capillary tubes (Microslide 1099, VitroCom Inc; Mt Lakes, NJ) or loaded into
80 micron 2X-CEL chambers (Hamilton Thorne; Beverly, MA). Three fields per
sample were manually selected and analyzed using the Hamilton Thorne IVOS
computerized semen analyzer (Beverly, MA).
embryo cryopreservation
Two-cell embryos were cryopreserved in 1.5 M propylene glycol as described by
Glenister and Rall[29].
embryo transfer
Pseudopregnant CByB6F1/J mice of 9 to 13 weeks of age were used as embryo
transfer recipients. Four to 15 embryos were transferred into one oviduct of each
female as described by Nagy et al.[30].
statistical Analysis
Percentages were arcsine transformed[31], and treatment differences were detect-
ed by analysis of variance (ANOVA) and preplanned comparisons in JMP 6 (SAS
Institute, Cary, NC). For presentation, means and standard errors of percentages
are shown.
Acknowledgements
The authors are grateful for the discussion and insight provided by Shannon By-
ers, Sian Clements, Tom Gridley, Mary Ann Handel, John Kulik and Stanley
Leibo. In addition, they appreciate the assistance provided by the Reproductive
Sciences Service at The Jackson Laboratory. Their skill in mouse husbandry, as
well as performing mouse surgeries, in vitro fertilizations, and sperm recoveries
are second to none.
Author contributions
Conceived and designed the experiments: GCO MVW JSF RAT. Performed the
experiments: GCO JSF. Analyzed the data: GCO MVW JSF RAT. Contributed
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