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Cep55 regulates spindle organization and cell cycle progression in meiotic oocyte.

Scientific reports (2015-11-20)
Zhao-Yang Xu, Xue-Shan Ma, Shu-Tao Qi, Zhen-Bo Wang, Lei Guo, Heide Schatten, Qing-Yuan Sun, Ying-Pu Sun
ZUSAMMENFASSUNG

Cep55 is a relatively novel member of the centrosomal protein family. Here, we show that Cep55 is expressed in mouse oocytes from the germinal vesicle (GV) to metaphase II (MII) stages. Immuostaining and confocal microscopy as well as time lapse live imaging after injection of mRNA encoding fusion protein of Cep55 and GFP identified that Cep55 was localized to the meiotic spindle, especially to the spindle poles at metaphase, while it was concentrated at the midbody in telophase in meiotic oocytes. Knockdown of Cep55 by specific siRNA injection caused the dissociation of γ-tubulin from the spindle poles, resulting in severely defective spindles and misaligned chromosomes, leading to metaphase I arrest and failure of first polar body (PB1) extrusion. Correspondingly, cyclin B accumulation and spindle assembly checkpoint (SAC) activation were observed in Cep55 knockdown oocytes. Our results suggest that Cep55 may act as an MTOC-associated protein regulating spindle organization, and thus cell cycle progression during mouse oocyte meiotic maturation.

MATERIALIEN
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Produktbeschreibung

Sigma-Aldrich
Tyrode′s Solution, Acidic, liquid, sterile-filtered, suitable for mouse embryo cell culture
Sigma-Aldrich
Anti-γ-Tubulin-Antikörper, monoklonaler Antikörper der Maus in Maus hergestellte Antikörper, clone GTU-88, ascites fluid