The Histone Deacetylase Inhibitor, CI994, Improves Nuclear Reprogramming and In Vitro Developmental Potential of Cloned Pig Embryos

Abstract

Epigenetic reprogramming and somatic cell nuclear transfer (SCNT) cloning efficiency were recently enhanced using histone deacetylase inhibitors (HDACis). In this study, we investigated the time effect of CI994, an HDACi, on the blastocyst formation rate, acetylation levels of H3K9 and H4K12, DNA methylation levels of anti-5-methylcytosine (5mC), and some mRNA expression of pluripotency-related genes in pig SCNT embryos. Treatment with 10 μM CI994 for 24 hours significantly improved the blastocyst formation rate of SCNT embryos in comparison with the untreated group (p < 0.05). Moreover, average fluorescence intensities of H3K9 and H4K12 in CI994-treated embryos were remarkably increased at the pseudo-pronuclear stage, but not at the blastocyst stage. The intensity of POU5F1 was higher in CI994-treated blastocysts than in control blastocysts, whereas that of 5mC did not differ between the two groups. The percentage of apoptotic cells in blastocysts was significantly higher in the untreated group than in the CI994-treated group. mRNA levels of POU5F1 and SOX2 were significantly increased in the CI994-treated group. These observations suggest that optimum exposure (10 μM for 24 hours) to CI994 after activation elevates the level of histone acetylation and subsequently improves the in vitro development of pig SCNT embryos.

Effects of resveratrol on in vitro maturation of porcine oocytes and subsequent early embryonic development following somatic cell nuclear transfer

“…Porcine somatic cell nuclear transfer (SCNT) is a required tool for human disease modelling to study pig‐to‐human xenotransplantation and for the improvement and breeding of disease‐resistant pig strains (Nagashima & Matsunari, ; Samiec & Skrzyszowska, ). The efficiency of SCNT in pigs is affected, to the highest degree, by factors such as (a) nuclear (meiotic), epigenomic and cytoplasmic maturity of nuclear recipient oocytes (Samiec & Skrzyszowska, ; Wang et al, ; Xu, Qian, Si, Qu, & Li, ); (b) epigenetic reprogrammability of somatic cell‐inherited nuclear genome (Jin et al, , ; Samiec & Skrzyszowska, ); (c) capability of nuclear‐transferred (NT) oocytes to be artificially activated (De Macedo et al, ; Samiec & Skrzyszowska, , ); (d) intergenomic communication between nuclear and mitochondrial compartments in NT oocytes and resultant cloned embryos (Estrada et al, ; Opiela, Samiec, & Romanek, ; Samiec, , ); and (e) incidence of apoptotic cell death in nuclear donor cells and cloned embryos (Ju, Rui, Lu, Lin, & Guo, ; Lee et al, ; Samiec & Skrzyszowska, ). In this regard, oocyte maturation at the nuclear, epigenomic and cytoplasmic levels seems to play a pivotal role in determining molecular quality and suitability of nuclear recipient oocytes for SCNT procedure (Jeon et al, ; Lin, Oqani, Lee, Shin, & Jin, ).…”

The HDAC inhibitor CI-994 acts as a molecular memory aid by facilitating synaptic and intra-cellular communication after learning

“…We focused on H3K27ac, a known marker of active enhancers that is enriched at activity-dependent regulatory elements after neuronal activation (30, 42, 52-54), correlates with gene transcription (30,55) and often co-occurs with H3K9ac, a marker of active promoters (56). Furthermore, in line with previous studies (15,25,57,58), we found that HDACi treatment increased global H3K27ac, alongside H3K9ac and H4K12ac as revealed by western blotting (Two-way ANOVA, F(3,60) = 22.47, P = 1.11e-13) (Supplemental Fig. 11).…”

Kidney ECM Pregel Nanoarchitectonics for Microarrays to Accelerate Harvesting Gene-Edited Porcine Primary Monoclonal Spheres

“… 35 The activity of somatic cells undergoing such an operation is significantly reduced, and their nuclear reprogramming ability is affected. 36 Furthermore, it will cause the surrogate sow to miscarry and the fetus to be deformed. Therefore, when preparing porcine-modified somatic cells, it is generally required to use high-activity cells with early generation and short cumulative culture times.…”