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. 2012 Jun 25;197(7):897–905. doi: 10.1083/jcb.201203062

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Copyright © 2012 by the Rockefeller University Press

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Figure 3. — Human sperm bind to huZP2 rescue eggs. (A) Human sperm binding to human ZP1 (huZP1), huZP2, and huZP3 rescue and huZP4 transgenic eggs. Noninseminated human oocytes (human) and Zp3^EGFP transgenic eggs (green zona) were positive and negative controls, respectively. Images obtained by confocal and DIC microscopy after staining with Hoechst were merged. Insets show magnification of a single humanized egg. (B) Same as A but with mouse sperm. Positive and negative controls were Zp3^EGFP transgenic eggs and two-cell embryos, respectively. (C) 3D reconstruction of a z stack of confocal images of human sperm binding to the huZP2 rescue egg. Sperm heads binding to the zona pellucid were visualized by Hoechst staining, and the projection of sperm tails into the media was detected by single wavelength reflection. (D) Same as A but with human ZP1 (huZP1), huZP2, and huZP3 transgenic eggs before crossing into the corresponding null background. (E) Human sperm binding to huZP2 rescue eggs after sperm capacitation for 30 min, 2 h, 3 h, and 4 h. Confocal z projections (top) and DIC (bottom) images after staining with Hoechst.