Expression and characterization of three Aurora kinase C splice variants found in human oocytes

• Published in: Molecular human reproduction Vol. 21; no. 8; pp. 633 - 644
• Main Authors: Fellmeth, Jessica E, Gordon, Derek, Robins, Christian E, Scott, Jr, Richard T, Treff, Nathan R, Schindler, Karen
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 01.08.2015
• Subjects: Aurora Kinase C - genetics; Cells, Cultured; Cumulus Cells - metabolism; Female; Fertility - genetics; Fertility - physiology; Gene Expression Regulation, Enzymologic; Humans; Immunoblotting; Immunohistochemistry; Meiosis - genetics; Meiosis - physiology; Oocytes - enzymology; Polymerase Chain Reaction; RNA Splicing - genetics; meiosis; oocyte maturation; female fertility; Aurora kinase C
• ISSN: 1360-9947; 1460-2407
• DOI: 10.1093/molehr/gav026

Chromosome segregation is an extensively choreographed process yet errors still occur frequently in female meiosis, leading to implantation failure, miscarriage or offspring with developmental disorders. Aurora kinase C (AURKC) is a component of the chromosome passenger complex and is highly expressed in gametes. Studies in mouse oocytes indicate that AURKC is required to regulate chromosome segregation during meiosis I; however, little is known about the functional significance of AURKC in human oocytes. Three splice variants of AURKC exist in testis tissue. To determine which splice variants human oocytes express, we performed quantitative real-time PCR using single oocytes and found expression of all three variants. To evaluate the functional differences between the variants, we created green fluorescent protein-tagged constructs of each variant to express in oocytes from Aurkc(-/-) mice. By quantifying metaphase chromosome alignment, cell cycle progression, phosphorylation of INCENP and microtubule attachments to kinetochores, we found that AURKC_v1 was the most capable of the variants at supporting metaphase I chromosome segregation. AURKC_v3 localized to chromosomes properly and supported cell cycle progression to metaphase II, but its inability to correct erroneous microtubule attachments to kinetochores meant that chromosome segregation was not as accurate compared with the other two variants. Finally, when we expressed the three variants simultaneously, error correction was more robust than when they were expressed on their own. Therefore, oocytes express three variants of AURKC that are not functionally equivalent in supporting meiosis, but fully complement meiosis when expressed simultaneously.