Trichostatin A affects histone acetylation and gene expression in porcine somatic cell nucleus transfer embryos.
Epigenetic aberrancies likely preclude correct and complete nuclear reprogramming after somatic cell nucleus transfer (SCNT) and may underlie the observed reduced viability of cloned embryos. In the current study, we tested the effects of the histone deacetylase-inhibitor trichostatin A (TSA) on preimplantation development and on histone acetylation and the gene expression of nucleus transfer (NT) porcine (Sus scrofa) embryos. Our results showed that 5 nM TSA for 26 h after reconstitution resulted in embryos (NTTSA) that reached the blastocyst stage at a higher level (48.1% vs. 20.2%) and increased number of cells (105.0 vs. 75.3) than that of the control (NTC) embryos. In addition, and unlike the NTC embryos, the treated embryos displayed a global acetylated histone H4 at lysine 8 profile similar to the in vitro-fertilized (IVF) and cultured embryos during the preimplantation development. Finally, we determined that several transcription factors exert a dramatic amount of genetic control over pluripotency, including Oct4, Nanog, Cdx2, and Rex01, the imprinting genes Igf2 and Igf2r, and the histone deacetyltransferase gene Hdac2. The NT blastocysts showed similar levels of Oct4, Cdx2, and Hdac2 but lower levels of Nanog than those of the IVF blastocyst. However, the NTTSA blastocysts showed similar levels of Rex01, Igf2, and Igf2r as those of IVF blastocysts, whereas the NTC blastocysts showed significantly lower levels for those genes. Our results suggest that TSA improves porcine SCNT preimplantation development and affects the acetylated status of the H4K8, rendering acetylation levels similar to those of the IVF counterparts.
Cervera
R P
RP
Cellular Reprogramming Laboratory, Centro de Investigación Príncipe Felipe, Valencia, Spain. rcervera@cipf.es
Martí-Gutiérrez
N
N
Escorihuela
E
E
Moreno
R
R
Stojkovic
M
M
D000107:Acetylation; D000818:Animals; D002467:Cell Nucleus / Q000187:drug effects* / Q000378:metabolism; D019976:Cloning, Organism; D046149:Embryo Culture Techniques; D004624:Embryo Transfer; D004622:Embryo, Mammalian / Q000187:drug effects / Q000378:metabolism; D047108:Embryonic Development / Q000187:drug effects; D044127:Epigenesis, Genetic / Q000187:drug effects; D015870:Gene Expression / Q000187:drug effects; D006657:Histones / Q000378:metabolism*; D006877:Hydroxamic Acids / Q000494:pharmacology*; D008239:Lysine / Q000378:metabolism; D053652:Nuclear Transfer Techniques; D034421:Sus scrofa / Q000196:embryology*; D014157:Transcription Factors / Q000502:physiology
D006657:Histones; D006877:Hydroxamic Acids; D014157:Transcription Factors; C012589:trichostatin A; D008239:Lysine