Abstract

KIF20A (Kinesin-like family member 20A), also called mitotic kinesin-like proteins 2 (MKLP2), is a mammalian mitotic kinesin-like motor protein of the Kinesin superfamily proteins (KIFs), which was originally involved in Golgi apparatus dynamics and thought to essential for cell cycle regulation during successful cytokinesis. In the present study, we investigated whether KIF20A has roles on porcine oocyte meiotic maturation and subsequent early embryo development. By immunofluorescence staining, KIF20A was found to exhibit a dynamic localization pattern during meiosis. KIF20A was restricted to centromeres after germinal vesicle breakdown (GVBD), transferred to the midbody at telophase I (TI), and again associated with centromeres at metaphase II (MII). Inhibition of endogenous KIF20A via a specific inhibitor, Paprotrain, resulted in failure of polar body extrusion. Further cell cycle analysis showed that the percentage of oocytes that arrested at early metaphase I (MI) stage increased after KIF20A activity inhibition; however, the proportion of oocytes at anaphase/telophase I (ATI) and MII stages decreased significantly. Our results also showed that KIF20A inhibition did not affect spindle morphology. In addition, KIF20A was localized at the nucleus of early embryos, and KIF20A inhibition resulted in failure of early parthenogenetic embryo development. These results demonstrated that KIF20A is critical for porcine oocyte meiotic maturation and subsequent early embryo development.

Highlights

  • To maintain genomic stability, normal multiplication of animal cell must undergo replication of the genome, chromosome segregation and subsequent cytokinesis

  • The subcellular localization of KIF20A was examined at different stages of meiotic maturation by immunofluorescent staining with a KIF20A antibody

  • KIF20A exhibited a dynamic localization pattern during meiosis. It localized at the centromeres in early metaphase I (MI) stage, and moved to the midbody in dividing oocytes during telophase I (TI); KIF20A again accumulated at centromeres at metaphase II (MII)

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Summary

Introduction

Normal multiplication of animal cell must undergo replication of the genome, chromosome segregation and subsequent cytokinesis. Cytokinesis is achieved by separating of sister chromatids to ensure faithful segregation of the chromosomal content [1]. The oocyte divides to form a highly polarized large MII arrested oocyte with a tiny first polar body. This process is essential for retention of maternal components for early embryo development [2]. Mitosis begins with the separation of centrosomes at prophase to ensure accurate chromosomal movement and segregation [3,4]. The central spindle, formed between segregating chromosomes during anaphase, is essential for alignment and accurate segregation of chromosomes. Results from several studies showed that MKLPs associated with the central spindle are required for proper cytokinesis [9,10,11]

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