PMID- 21753193 OWN - NLM STAT- MEDLINE DCOM- 20120320 LR - 20121115 IS - 1529-7268 (Electronic) IS - 0006-3363 (Linking) VI - 85 IP - 5 DP - 2011 Nov TI - Histone deacetylase inhibition improves activation of ribosomal RNA genes and embryonic nucleolar reprogramming in cloned mouse embryos. PG - 1048-56 LID - 10.1095/biolreprod.110.089474 [doi] AB - Our group found that the treatment of embryos with histone deacetylase inhibitors (HDACi), including trichostatin A, Scriptaid, suberoylanilide hydroxamic acid, and oxamflatin, after cloning by somatic cell nuclear transfer (SCNT) resulted in significantly improved efficiency. Although many researchers have investigated the use of HDACi treatment to improve the quality of cloned mouse embryos, the mechanism underlying this treatment has not been completely understood. We believe that the effect of HDACi on embryonic gene activation (EGA) is important for normal development of cloned embryos. In the present study, using highly sensitive fluorescence in situ hybridization (FISH) with probes complementary to mouse rDNA, the effect of Scriptaid on the onset of rRNA synthesis was examined in cloned embryos. In addition, to determine how Scriptaid affects pre-rRNA processing machinery in SCNT embryos with activated rDNA transcription, functional nucleolar formation was analyzed in detail by combined assessment of rRNA synthesis and nucleolar protein allocation in preimplantation embryos. In this experiment, at least part of the rRNA localization by FISH was substituted by 5-bromouridine 5'-triphosphate staining after alpha-amanitin treatment. The results show that in the late 2-cell stage, a number of SCNT embryos initiated transcriptional activation while having one blastomere showing inactivated rRNA transcription and another blastomere showing activated rRNA transcription and despite both nuclei being in interphase. In addition, in some SCNT embryos, the same nuclei contained a mixture of inactively and actively transcribed rRNA, which was rarely observed in intracytoplasmic sperm injection embryos. This asynchronous transcription induced a delay of one cell cycle in SCNT embryo activation of functional nucleoli. Scriptaid can overcome this failure in the timely onset of embryonic gene transcription by activation of rRNA genes and promotion of nucleolar protein allocation during the early phase of EGA. FAU - Bui, Hong-Thuy AU - Bui HT AD - Department of Animal Biotechnology, College of Animal Bioscience and Biotechnology/Animal Resources Research Center, Konkuk University, Seoul, Republic of Korea. FAU - Seo, Hyeon-Jeong AU - Seo HJ FAU - Park, Mi-Rung AU - Park MR FAU - Park, Jong-Yi AU - Park JY FAU - Thuan, Nguyen Van AU - Thuan NV FAU - Wakayama, Teruhiko AU - Wakayama T FAU - Kim, Jin-Hoi AU - Kim JH LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20110713 PL - United States TA - Biol Reprod JT - Biology of reproduction JID - 0207224 RN - 0 (Enzyme Inhibitors) RN - 0 (Hydroxamic Acids) RN - 0 (Hydroxylamines) RN - 0 (Quinolines) RN - 0 (RNA, Ribosomal) RN - 0 (oxamflatin) RN - 0 (scriptaid) RN - 3X2S926L3Z (trichostatin A) RN - EC 3.5.1.98 (Histone Deacetylases) SB - IM MH - Animals MH - Cell Nucleolus/*physiology MH - Cloning, Organism/methods MH - Embryonic Development/*physiology MH - Enzyme Inhibitors/*pharmacology MH - Histone Deacetylases/*drug effects MH - Hydroxamic Acids/pharmacology MH - Hydroxylamines/pharmacology MH - Mice MH - Mice, Inbred C57BL MH - Mice, Inbred DBA MH - Mice, Inbred Strains MH - Models, Animal MH - *Nuclear Transfer Techniques MH - Quinolines/pharmacology MH - RNA, Ribosomal/*genetics MH - Transcriptional Activation/physiology EDAT- 2011/07/15 06:00 MHDA- 2012/03/21 06:00 CRDT- 2011/07/15 06:00 PHST- 2011/07/15 06:00 [entrez] PHST- 2011/07/15 06:00 [pubmed] PHST- 2012/03/21 06:00 [medline] AID - biolreprod.110.089474 [pii] AID - 10.1095/biolreprod.110.089474 [doi] PST - ppublish SO - Biol Reprod. 2011 Nov;85(5):1048-56. doi: 10.1095/biolreprod.110.089474. Epub 2011 Jul 13.