Analysis of human Aurora C Kinase variants in oocyte meiosis

The birth of a healthy child is often a priority for couples, yet many couples will unfortunately suffer from infertility. Aneuploidy is the leading genetic abnormality that causes infertility. Aneuploidies in sperm and eggs arise from errors during chromosome segregation in meiosis. Though many of the critical players are known and mutants characterized, the regulatory components involved in directing accurate chromosome segregation are still not entirely clear. Aurora kinase C is one of the critial players in chromosomes segregation in meiosis and in this thesis, I establish the use of Aurkc-/- mouse oocytes as an effective model for studying human AURKC ex vivo. By comparing the rescue of the knockout phenotype when hAURKC and mAURKC are expressed in the oocytes, I show that hAURKC functions similarly in female meiosis as does mAURKC. I also characterized three splice variants of hAURKC in this model after establishing their simultaneous expression in human oocytes. The three splice variants have different abilities and stabilities in meiosis, but promote optimal chromosome segregation when expressed simultaneously during oocyte meiosis. Finally, I used the verified model to characterize the phenotype of three sterility-associated mutations in human AURKC. I show through functional assays that the three mutations have differing abilities in meiosis, but all result in a loss of function of the AURKC-CPC, which causes meiotic arrest and polyploid gametes. This work contributes to the field of reproductive biology by providing a tractable model for studying human AURKC and for further analysis of mutants and variants.Ph.D.Includes bibliographical referencesby Jessica Fellmet