title: Altering histone acetylation status in donor cells with suberoylanilide hydroxamic acid does not affect dog cloning efficiency.
abstract: Although dog cloning technology has been applied to conservation of endangered canids, propagation of elite dogs, and production of transgenic dogs, the efficiency of cloning is still very low. To help overcome this problem, we evaluated the effect of treating donor cells with suberoylanilide hydroxamic acid (SAHA), a histone deacetylase inhibitor, on dog cloning efficiency. Relative messenger RNA expressions of the bax1/bcl2 ratio and Dnmt1 in fibroblasts treated with different concentrations (0, 1, 10, 50 μM) of SAHA and durations (0, 20, 44 hours) were compared. Treatment with 1 μM for 20 hours showed significantly lower bax1/bcl2 and Dnmt1 transcript abundance. Acetylation of H3K9 was significantly increased after SAHA treatment, but H4K5, H4K8 and H4K16 were not changed. After SCNT using control or donor cells treated with SAHA, a total of 76 and 64 cloned embryos were transferred to seven and five recipients, respectively. Three fetuses were diagnosed in both control and SAHA-treated groups by ultrasonography 29 days after the embryo transfer, but there was no significant difference in the pregnancy rate (4.2% vs. 4.3%). In conclusion, although SAHA treatment as used in this study significantly decreased bax1/bcl2 and Dnmt1 transcripts of donor nuclei, as well as increased H3 acetylation, it was not enough to increase in vivo developmental competence of cloned dog embryos.
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Gwanak-gu, Seoul, Republic of Korea.
authors.forename: Min Jung
authors.initials: MJ
authors.lastname: Kim
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Gwanak-gu, Seoul, Republic of Korea.
authors.forename: Hyun Ju
authors.initials: HJ
authors.lastname: Oh
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Gwanak-gu, Seoul, Republic of Korea.
authors.forename: Geon A
authors.initials: GA
authors.lastname: Kim
authors.affiliation: Department of Veterinary Physiology, College of Veterinary Medicine and Research Institute for Veterinary Science, Seoul National University, Gwanak-gu, Seoul, Republic of Korea.
authors.forename: Han Na
authors.initials: HN
authors.lastname: Suh
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Gwanak-gu, Seoul, Republic of Korea.
authors.forename: Young Kwang
authors.initials: YK
authors.lastname: Jo
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Gwanak-gu, Seoul, Republic of Korea.
authors.forename: Yoo Bin
authors.initials: YB
authors.lastname: Choi
authors.affiliation: Animal Biotechnology Division, National Institute of Animal Science, Wanju-gun, Jeollabuk-do, Korea.
authors.forename: Dong Hoon
authors.initials: DH
authors.lastname: Kim
authors.affiliation: Department of Veterinary Physiology, College of Veterinary Medicine and Research Institute for Veterinary Science, Seoul National University, Gwanak-gu, Seoul, Republic of Korea.
authors.forename: Ho Jae
authors.initials: HJ
authors.lastname: Han
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Gwanak-gu, Seoul, Republic of Korea. Electronic address: bclee@snu.ac.kr.
authors.forename: Byeong Chun
authors.initials: BC
authors.lastname: Lee
mesh_terms: D000107:Acetylation; D000818:Animals; D017209:Apoptosis / Q000235:genetics; D065150:Cellular Reprogramming / Q000235:genetics; D019976:Cloning, Organism / Q000379:methods / Q000662:veterinary*; D004285:Dogs / Q000196:embryology*; D004305:Dose-Response Relationship, Drug; D004624:Embryo Transfer / Q000662:veterinary; D047108:Embryonic Development / Q000187:drug effects; D056727:Endangered Species; D005260:Female; D005347:Fibroblasts / Q000737:chemistry / Q000187:drug effects / Q000648:ultrastructure; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006657:Histones / Q000378:metabolism; D006877:Hydroxamic Acids / Q000008:administration & dosage / Q000494:pharmacology*; D053652:Nuclear Transfer Techniques / Q000662:veterinary; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis; D000077337:Vorinostat
chemical_list: D056572:Histone Deacetylase Inhibitors; D006657:Histones; D006877:Hydroxamic Acids; D012333:RNA, Messenger; D000077337:Vorinostat
keywords: Cloning; Dog; Histone deacetylase inhibitor; SCNT; Suberoylanilide hydroxamic acid