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Figure 2


Fig. 2. Parthenogenesis follows normal meiotic progression in rPLCZ1-expressing hemizygous female mice. (A) Immunofluorescence microscopy of oocytes matured in vitro at <2 (GV) or 8-10 (mI) hours after meiotic resumption, or ~13 hours post-hCG (mII). Oocytes were from hemizygotes (CS) and age-matched non-transgenic littermates (wt). TUBA2 labelling is green and genomic DNA red. Arrows and arrowheads respectively mark the first polar body (Pb1) and mII plate. (B) Proportion of age-matched oocytes upon collection at mII ~13 (white) and 24 (grey) hours post-hCG. (C) Hofmann image of F4 oocytes 13.5-14 hours post-hCG, showing metaphase or anaphase-telophase distortions (arrowheads) and Pb1 (arrows). (D) Immunofluorescence microscopy of different oocytes from C, ~14 hours post-hCG, at early (upper) and late (beneath) stages of spindle rotation, represented diagrammatically to the right. Staining and key to arrow and arrowheads are as for A. (E) Ratiometric Fura 2-AM [Ca2+]i imaging of representative oocytes at different stages, performed as for A but with mII oocytes 14.5 hours post-hCG. Oocytes were from non-transgenic control (wt, upper) and age-matched transgenic (CS, beneath) females. (F,G) Hofmann images (F) and bar chart (G) showing development in vitro of CS16 F3 (CS) and SrCl2-induced wild-type (wt) haploid parthenogenotes at the times shown after oocyte collection or activation. Pronuclei and Pb2 in F are respectively indicated with arrowheads and arrows. (H) Preimplantation development following nuclear transfer (nt) from CS16 F4 cumulus cell nuclei into enucleated wild-type oocytes without exogenous activation, shown at the times indicated (hours) post-nt. Pronuclei are indicated with arrowheads. (I) Developmental rates in vitro following nt of CS16 F3 and F4, or age-matched non-transgenic or pCAG->mtVenus transgenic (Shoji et al., 2006) cumulus cells (control) into enucleated wild-type oocytes. m/b (in G,I), morula/blastocyst. Scale bars: 20 µm.





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