Paternal Maternal crosstalk: How the sperm chromatin is handled by egg factors at fertilisation.

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Sperm is responsible for delivering the paternal genome to the embryo. However, it also contributes additional epigenetic cues that have the capacity to influence development. To decipher the mechanism of paternally derived transmission of information to the embryo, we aimed to identify the maternal factors that are responsible for the remodelling of sperm chromatin at fertilisation. To this end, we employed the Xenopus model, in which egg extract can be utilised to emulate the event occurring at fertilisation. Permeabilised sperm nuclei are incubated in egg extract to enable paternal chromatin remodelling, and replication, by maternal factors. Control sample sets were generated using spermatid, the immediate precursor of sperm during spermiogenesis, to pinpoint sperm-specific binding event. Additionally, replication inhibition was carried out to define replication-independent binding events. Egg factors bound to paternal chromatin were identified and differentially quantified by dia-PASEF analysis. Our analysis captured known events and yielded score of new interactors including chromatin modifiers as well as RNA regulators. Binding of selected candidates (i.e. HDAC1, DDPA4, HMBOX1) was confirmed through western blot analysis. Functional relevance of egg factor binding to sperm chromatin will next be evaluated via trim-away depletion in frog and bovine embryos.

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