Figure 1.
Subcellular localization of KIF20A in porcine oocytes.
Immunofluorescent staining was employed to show the subcellular localization of KIF20A. KIF20A exhibited a dynamic localization pattern during porcine oocyte meiotic maturation. KIF20A localized at centromeres at the germinal vesicle breakdown (GVBD) stage and metaphase I (MI) stage, then transferred to the midbody in dividing oocytes during telophase (TI), finally, KIF20A was accumulated at centromeres again at metaphase II (MII). Red, KIF20A; blue, chromatin.
Figure 2.
Disrupting KIF20A activity inhibits the meiosis competence and cell cycle progression of porcine oocytes.
(A) Granulosa cell diffusion of COCs was inhibited during meiosis. Almost all of COCs in control groups were surrounded by more than 5 layers of intact cumulus cells, whereas the COCs in 10, 20 and 50 µM Paprotrain treatment groups showed weakened diffusion. (B) Rates of first polar body extrusion of porcine oocytes after 44 h culture with different treatments (Control, 10 µM, 20 µM, and 50 µM). Oocytes failed to extrude the first polar body after treatment with KIF20A inhibitor. * significant difference (p<0.05). (C) Effects of KIF20A disruption on cell cycle during oocyte meiosis maturation. The cell cycle progression of porcine oocytes was disturbed during meiosis. The oocytes arrested at GVBD or MI stage and cannot perform the proper progression of cell cycle after KIF20A disruption. * significant difference (p<0.05).
Figure 3.
Effects of KIF20A inhibition on spindle structure and chromosome alignment of porcine oocytes.
(A) Normal Spindle structure and chromosome alignment were investigated after KIF20A inhibition. (B) Rates of abnormal oocytes (displaying chromosome misalignment and aberrant spindles) after KIF20A inhibition.
Figure 4.
Localization of KIF20A in porcine early embryo.
Samples were collected from zygote the to the 4-cell stage. KIF20A was primarily accumulated at the nucleus from the zygote to the 4-cell embryos stages. Green, KIF20A; blue, chromatin. Bar = 20 µm.
Figure 5.
KIF20A inhibition affects the divisions of embryos.
(A) Schematic diagram of cleavage of early embryos in different groups. The black arrow indicated a 4-cell embryo in control group.(B) Effect of KIF20A inhibition on cleavage ability of zygotes. The embryonic development rate has dropped after KIF20A activity inhibition. Almost all of the embryos cleaved normally in the control groups, while the cleavage of embryos was blocked in treatment groups, especially in the 50 µM groups. * significant difference (p<0.05).