Sperm histone code and epigenetic-mediated subfertility

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It is now generally accepted that a sperm-specific epigenetic code is established during spermiogenesis and transmitted to the oocyte. Aberrations may represent one reason for idiopathic male infertility resulting in decreased IVF/ICSI outcome. In a previous study, gene promoters associated with histone H4 acetylated at lysine 12 (H4K12ac) and also exonic sequences associated with histone H3 acetylated at lysine 9 (H3K9ac) have been identified in ejaculated spermatozoa from fertile donors and subfertile patients applying ChlP-on-chip HG18 promoter array and ENCODE array, respectively. In the present study, the following aims will be addressed: 1) To clarify whether H4K12ac-associated gene promoters colocalize with other histones, such as H3K9ac, H3K4me3 and H3K27me3, or protamine applying Re-ChIP; 2) To determine binding sequences for protamine-1 iri fertile donors and subfertile patients; 3) To analyze the methylation status of CpG dinucleotides within gene promoters of developmental genes associated with H4K12ac and protamine-1 applying pyrosequencing; 4) To analyze the possible role of H4K12ac and H3K9ac on paternal imprinted loci; 5) To analyze expression of H4K12ac-associated genes in the paternal pronucleus of mouse zygote using in-situ hybridization and immunofluorescence in combination with confocal microscopy; 6) To perform a genome-wide bioinformatic-based data comparision of own data from H4K12ac ChlP-on-chip and protamine-1 ChlPsequencing with transcriptomes from human preimplantation embryos/embryonic stem cells and data generated in bovine model providing paternal contributed candidate genes for eariy development (coop, with project 7).