Abstract

SETD2 (SET domain containing protein 2) acts as a histone H3 lysine 36 (H3K36)-specific methyltransferase and may play important roles in active gene transcription in human cells. However, its expression and role in porcine oocytes and preimplantation embryos are not well understood. Here, we used immunofluorescence and laser scanning confocal microscopy to examine SETD2 expression in porcine fetal fibroblasts, oocytes, and preimplantation embryos derived from in vitro fertilization (IVF), parthenogenetic activation (PA), and somatic cell nuclear transfer (SCNT). In porcine fetal fibroblasts, SETD2 expression was detected in interphase cells, but not in M (mitotic)-phase cells. The SETD2 signal was observed in non-surrounded nucleolus (NSN)-stage oocytes, but not in surrounded nucleolus (SN)-, metaphase I (MI)-, or metaphase II (MII)-stage oocytes. The SETD2 signal was detectable in sperm, and undetectable immediately after fertilization, detectable at the 2-cell stage, and peaked at the 4-cell stage of IVF embryos in which porcine embryonic genome is activated. Similar to the pattern found in IVF embryos, the SETD2 signal was absent from PA embryos at the 1-cell stage, but it was detected at the 2-cell stage and thereafter maintained to the blastocyst stage. Interestingly, unlike the IVF and PA embryos, the SETD2 signal was detected throughout the development of SCNT embryos, including at the 1-cell stage. These data suggest that SETD2 may be functional for embryonic gene transcription in porcine preimplantation embryos. It is further speculated that the aberrant expression of SETD2 at the 1-cell stage of porcine SCNT embryos may be a factor in the low efficiency of cloning in pig.

Highlights

  • Histone modifications, including acetylation, phosphorylation, methylation, and ubiquitination, etc., are connected with many fundamental biological processes in cells [1]

  • SETD2 signal in porcine oocyte and embryo always detected in non-surrounded nucleolus (NSN)-stage oocytes (100%, 12 of 12), whereas little (9.1%, 1 of 11) or no such signal (90.9%, 10 of 11) was detected in surrounded nucleolus (SN)-stage oocytes (Fig 2A and 2B)

  • We examined the SETD2 status in parthenogenetic activation (PA) embryos using an antibody against SETD2

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Summary

Introduction

Histone modifications, including acetylation, phosphorylation, methylation, and ubiquitination, etc., are connected with many fundamental biological processes in cells [1]. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Methods
Results
Conclusion
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