Acetylation and methylation profiles of H3K27 in porcine embryos cultured in vitro

Abstract

Methylation and acetylation of histone H3 at lysine 27 (H3K27) regulate chromatin structure and gene expression during early embryo development. While H3K27 acetylation (H3K27ac) is associated with active gene expression, H3K27 methylation (H3K27me) is linked to transcriptional repression. The aim of this study was to assess the profile of H3K27 acetylation and methylation (mono-, di- and trimethyl) during oocyte maturation and early development in vitro of porcine embryos. Oocytes/embryos were fixed at different developmental stages from germinal vesicle to day 8 blastocysts and submitted to an immunocytochemistry protocol to identify the presence and quantify the immunofluorescence intensity of H3K27ac, H3K27me1, H3K27me2 and H3K27me3. A strong fluorescent signal for H3K27ac was observed in all developmental stages. H3K27me1 and H3K27me2 were detected in oocytes, but the fluorescent signal decreased through the cleavage stages and rose again at the blastocyst stage. H3K27me3 was detected in oocytes, in only one pronucleus in zygotes, cleaved-stage embryos and blastocysts. The nuclear fluorescence signal for H3K27me3 increased from the 2-cell stage to 4-cell stage embryos, decreased at the 8-cell and morula stages and increased again in blastocysts. Different patterns of the H3K27me3 mark were observed at the blastocyst stage. Our results suggest that changes in the H3K27 methylation status regulate early porcine embryo development as previously shown in other species.

Comprehensive histochemical profiles of histone modification in male germline cells during meiosis and spermiogenesis: Comparison of young and aged testes in mice

“…anti-H3K4me2 rabbit polyclonal 1/500 Millipore (07-030) [13][14][15] anti-H3K4me3 rabbit polyclonal 1/500 Abcam (ab8580) [16][17][18] anti-H3K9me3 rabbit polyclonal 1/500 Abcam (ab8898) [19][20][21] anti-H3K27ac rabbit polyclonal 1/500 Abcam (ab4729) [22][23][24] anti-H3K27me2 rabbit polyclonal 1/500 Abcam (ab24684) [25,26] anti-H3K27me3 rabbit polyclonal 1/500 Millipore (07-449) [24,[27][28][29] anti-H3K79me2 rabbit polyclonal 1/500 Abcam (ab3594) [30][31][32] anti-H3K79me3 rabbit polyclonal 1/500 Abcam (ab2621) [33][34][35] anti-SCP3 mouse monoclonal 1/500 Abcam (ab97672) [36][37][38] anti-α-Tubulin (S2 Fig) mouse monoclonal 1/500 Sigma-Aldrich (T6199) [39,40] All antibodies are commercially available, and their specificity has been reported in previous studies.…”

The histone lysine demethylase KDM7A is required for normal development and first cell lineage specification in porcine embryos

“…Moreover, H3K9me3 is also important for repressing ICM-specific genes in the TE lineage [48]. Different levels of H3K27me3 were also observed between ICM and TE cells in pig blastocysts [49,50], which suggests a role in the first cell lineages specification. Findings from this study provide evidence that KDM7A participates in this process by regulating methylation levels on both H3K9 and H3K27.…”

Enhancement of the efficiency of oocyte vitrification through regulation of histone deacetylase 6 expression

“…Sperm chromatin is de-condensed followed by recondensation in the early fertilization, in which histone acetylation alters accordingly. Therefore, improper histone acetylation activity may affect the process of meiosis of oocytes, and subsequently affect preimplantation of the embryos [16][17][18]. In this study, we explored the involvement of HDAC6 in the regulation and control of oocyte survival and maintenance of developmental potential.…”