Effect of Hsp27 on early embryonic development in the mouse

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Presentation transcript:

Effect of Hsp27 on early embryonic development in the mouse Shan Liu, Xiaonan Dai, Lingbo Cai, Xiang Ma, Jinjuan Liu, Shiwen Jiang, Jiayin Liu, Yugui Cui Reproductive BioMedicine Online Volume 26, Issue 5, Pages 491-499 (May 2013) DOI: 10.1016/j.rbmo.2013.01.005 Copyright © 2013 Reproductive Healthcare Ltd. Terms and Conditions

Figure 1 Localization of Hsp27 in zygotes and early embryos by immunofluorescence. Embryo were cultured in vitro for 0h (PN), 24h (2-cell), 48h (4-cell), 72h (8-cell, morula), 96h (blastocyst) and stained with specific Hsp27 antibody (green, visualized at 488nm) and propidium iodide (PI, red, visualized at 561nm) (scaling: X 0.26μm, Y 0.26μm; stack size: X 265.69μm, Y 265.69μm; pinhole: Ch1 90μm, Ch2 90μm). Bars=20μm. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.) Reproductive BioMedicine Online 2013 26, 491-499DOI: (10.1016/j.rbmo.2013.01.005) Copyright © 2013 Reproductive Healthcare Ltd. Terms and Conditions

Figure 2 Localization and expression of Hsp27 in 4-cell and 8-cell embryos after microinjection with AdCMV-Hsp27 or AdsiRNA-Hsp27 into zygotes. (A, E) Hsp27 in the control group. (B, F) Hsp27 in the AdCMV-Hsp27 group. (C, G) Hsp27 in the AdsiRNA-Hsp27 group; see legend for Figure 1 for staining and visualization details; bars=20μm. (D, H) Results of real-time RT-PCR showing Hsp27 expression in the AdCMV-Hsp27 and AdsiRNA-Hsp27 groups. Expression level was calculated from the Ct values by the 2(–Delta Delta Ct) method, and the mRNA ratio (arbitrary units) of Hsp27 was calculated with respect to that of controls; values are mean±SD; ∗P<0.05 versus control. Reproductive BioMedicine Online 2013 26, 491-499DOI: (10.1016/j.rbmo.2013.01.005) Copyright © 2013 Reproductive Healthcare Ltd. Terms and Conditions

Figure 3 Cell apoptosis of the mouse blastocyst derived from the zygotes microinjected with AdCMV-Hsp27 or AdsiRNA-Hsp27. (a–c) Control embryos. (d–f) Embryos cultured for 96h after microinjection with AdCMV-Hsp27. (g–i) Embryos cultured for 96h after microinjection with AdsiRNA-Hsp27. (j–l) Positive control blastocysts, treated with DNase before incubation with TUNEL reagents. Fragmented DNA were labelled by the TUNEL reaction (green) and chromatin content was determined by staining with propidium iodide (PI, red); merged stains appear yellow. Bars=20μm. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.) Reproductive BioMedicine Online 2013 26, 491-499DOI: (10.1016/j.rbmo.2013.01.005) Copyright © 2013 Reproductive Healthcare Ltd. Terms and Conditions

Figure 4 Evaluation of embryo quality. (A) Blastocyst morphology: blastocysts from up-regulated (AdCMV-Hsp27) and silenced (AdsiRNA-Hsp27) zygotes were subjected to nuclear Hoechest 33342 staining (blue) and the cell nuclei were counted; treated groups showed similar morphology to the controls. Bars=20μm. (B, C) Total cell number (B) and cell death index (C) per embryo were similar in the three groups. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.) Reproductive BioMedicine Online 2013 26, 491-499DOI: (10.1016/j.rbmo.2013.01.005) Copyright © 2013 Reproductive Healthcare Ltd. Terms and Conditions