bioRxiv Science⌕ Search

Biology subjects

Nikalayevich, E.

Publications and source records attributed to Nikalayevich, E..

2 recordsLinked to original sources

An interpretable and versatile machine learning approach for oocyte phenotyping

Meiotic maturation is a crucial step of oocyte development allowing its potential fertilization and embryo development. Elucidating this process is important both for fundamental research and assisted reproductive technology. However, only few computational tools, based on non-invasive measurements, are currently available to characterize oocyte meiotic maturation. Here, we develop a computational framework to phenotype oocytes based on images or movies acquired exclusively in transmitted light. We first trained neural networks to segment the contours of oocytes and their zona pellucida using a diverse cohort of both mouse and human oocytes. We then defined a comprehensive set of morphological features to describe a single oocyte. We have implemented these steps in a versatile and user-friendly open source Fiji plugin available to the mouse and human oocyte community. Then, we present a machine learning pipeline based on selected features to automatically recognize oocyte populations and determine their morphological differences. Its first application is a novel approach to screen oocyte strains and automatically identify their morphological characteristics. We demonstrate its potential by phenotyping a well characterized genetically modified mouse oocyte strain. Its second application is to predict and characterize the maturation potential of oocytes. Importantly, we identify two new features to assess mouse oocyte maturation potential, consisting in the texture of the zona pellucida and the cytoplasmic particles size. Eventually, we tested whether these mouse oocyte quality features were applicable to human oocytes developmental potential.

bioinformatics↗

Aurora B/C-dependent phosphorylation promotesRec8 cleavage in mammalian oocytes

To generate haploid gametes, cohesin is removed in a step-wise manner from chromosome arms in meiosis I and the centromere region in meiosis II, to segregate chromosomes and sister chromatids, respectively. Meiotic cohesin removal requires cleavage of the meiosis-specific kleisin subunit Rec8 by the protease Separase[1, 2]. In yeast, Rec8 is kept in a non-phosphorylated state by the action of PP2A-B56, which is localised to the centromere region, thereby preventing cohesin removal from this region in meiosis I[3-5]. However, it is unknown whether Rec8 has to be equally phosphorylated for cleavage, and whether centromeric cohesin protection is indeed brought about by dephosphorylation of Rec8 preventing cleavage, in mammalian meiosis. The identity of one or several potential Rec8-specific kinase(s) is also unknown. This is due to technical challenges, as Rec8 is poorly conserved preventing a direct translation of the knowledge gained from model systems such as yeast and C. elegans to mammals, and additionally, there is no turn-over of Rec8 after cohesion establishment, preventing phosphomutant analysis of functional Rec8. To address how Rec8 cleavage is brought about in mammals, we adapted a biosensor for Separase to study Rec8 cleavage in single mouse oocytes by live imaging, and identified phosphorylation sites promoting cleavage. We found that Rec8 cleavage by Separase depends on Aurora B/C kinase activity, and identified a residue promoting cleavage and being phosphorylated in an Aurora B/C kinase-dependent manner. Accordingly, inhibition of Aurora B/C kinase during meiotic maturation impairs endogenous Rec8 phosphorylation and chromosome segregation.

cell biology↗