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Effects of Plk1 on Meiotic Spindle Structure of Porcine (Sus scrofa) Oocytes During Metaphase Ⅰ Stage |
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Abstract Polo-like kinase 1 (Plk1) has been determined to have a variety of roles in the precise regulation of centrosome duplication and separation, spindle assembly, chromosome detachment, however, much less is known about its expression and function during the meiotic progression in oocytes. The present study was conducted to investigate the dynamic expression and the possible roles of Plk1 in the metaphase Ⅰ (MⅠ) stage spindle formation in porcine oocytes. Immunofluorescent staining coupled with confocal microscopy imaging techniques was used to examined the dynamic changes of Plk1. Then, the specific inhibitor GSK461364 was used to explore the potential functions of Plk1 during the meiotic maturation of porcine oocytes. Results showed that Plk1 was observed at germinal vesicle breakdown (GVBD), MⅠ and anaphase I and telophase I (ATI) stage, with a similar distribution of α-tubulin location. GSK461364 treatment resulted in the failure of polar body extrusion in pig oocytes in a concentration-dependent manner, and the first polar body emission rate was significantly decreased (39.37+5.46)% VS (81.10+7.69)% (P<0.01) when the concentration of GSK461364 reached 0.3 mol/L, which accompanied by misaligned chromosomes. Additional confocal microscopy results showed that GSK461364 treatment resulted in a significantly higher percentage of the treated oocytes with aberrant spindles during the first meiotic division. This study show that distribution of Plk1 is closely associated with the α-tubulin expression, and the regulation is related to its effects on spindle formation during the first meiotic division in porcine oocytes. The results of this study will provide an experimental basis for further understanding the regulation mechanism of oocyte meiosis and further improve the in vitro maturation efficiency of porcine oocytes.
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Received: 15 August 2017
Published: 25 March 2018
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