Fertility and Sterility Arsenite induces aberrations in meiosis that can be prevented by coadministration of N- acetylcysteine in mice  Paula A.A.S. Navarro, M.D., Ph.D., Lin Liu, Ph.D., Rui A. Ferriani, M.D., Ph.D., David L. Keefe, M.D.  Fertility and Sterility  Volume 85, Pages 1187-1194 (April 2006) DOI: 10.1016/j.fertnstert.2005.08.060 Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 1 Immunofluorescence images of spindles and chromosomes of CD-1 mouse oocytes from control (A) and in vitro arsenite-treated groups (B–D). (A) Chromosome alignment over the metaphase II plate of a normal spindle. (B) Chromosome misalignments over a disrupted meiosis I spindle 12–14 hours in culture after in vitro treatment with 2 μg/mL of arsenite for 2 hours. (C) Chromosome misalignments over a disrupted telophase I spindle 12–14 hours in culture after in vitro treatment with 2 μg/mL arsenite for 2 hours. (D) Severe chromosome spreading over a disrupted meiosis II spindle 15–17 hours in culture after in vitro treatment with 2 μg/mL arsenite for 2 hours. Green, meiosis spindles stained by anti-β-tubulin and fluorescein isothiocyanate–conjugated secondary antibody; blue, chromosomes stained by Hoechst 33342; red, actin-filaments. Scale bar = 10 μm. Navarro. Arsenite induces aberrations in meiosis. Fertil Steril 2006. Fertility and Sterility 2006 85, 1187-1194DOI: (10.1016/j.fertnstert.2005.08.060) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 2 PolScope imaging of CD-1 mouse oocytes from control (A) and in vitro arsenite-treated groups (B, C). (A) Normal meiosis II spindles. (B) Disrupted meiosis I spindles 12–14 hours in vitro maturation (IVM) culture after in vitro treatment with 2 μg/mL arsenite for 2 hours. (C) Disrupted meiosis II spindles at 15–17 hours in IVM culture after in vitro treatment with 2 μg/mL arsenite for 2 hours. Scale bar = 10 μm. Navarro. Arsenite induces aberrations in meiosis. Fertil Steril 2006. Fertility and Sterility 2006 85, 1187-1194DOI: (10.1016/j.fertnstert.2005.08.060) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 3 PolScope (upper panel) and immunofluorescence images (lower panel) of CD-1 mouse oocytes 12–14 hours in vitro maturation (IVM) culture. Control oocytes show normal meiosis II (MII) spindles and chromosome alignment at the metaphase plate. Oocytes treated with 2 μg/mL of arsenite (As) for 2 hours exhibit disrupted spindles and/or chromosome misalignment (aberrant meiosis I is shown). Oocytes treated with 2 μg/mL of arsenite plus 30 mM N-acetylcysteine (NAC) for 2 hours (As + NAC) display normal MII spindles and chromosome alignment at the metaphase plate, comparable to those of controls. Green, spindles stained by anti-β-tubulin and fluorescein isothiocyanate–conjugated secondary antibody; blue, chromosomes stained by Hoechst 33342. Scale bar = 10 μm. Navarro. Arsenite induces aberrations in meiosis. Fertil Steril 2006. Fertility and Sterility 2006 85, 1187-1194DOI: (10.1016/j.fertnstert.2005.08.060) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions