title: Trichostatin A modulates apoptotic-related gene expression and improves embryo viability in cloned bovine embryos.
abstract: Low efficiency of somatic cell nuclear transfer (SCNT) is attributed to incomplete reprogramming of transferred nuclei into oocytes. Trichostatin A (TSA), a histone deacetylase inhibitor, has been used to enhance nuclear reprogramming following SCNT. However, the molecular mechanism of TSA for the improvement of the preimplantation embryo and fetal development following SCNT is not known. The present study investigates embryo viability and gene expression of cloned bovine preimplantation embryos in the presence and absence of TSA compared to embryos produced by in vitro fertilization or parthenogenetic activation. Our results indicated that TSA treatment significantly improved total and inner cell mass (ICM) cell number and ratio of ICM:trophectoderm (TE) and also decreased the apoptotic index including total, ICM, and ratio of ICM:TE. Four apoptotic-related genes, Bcl-xL, survivin, Bcl2-associated X protein (Bax), and caspase 3 (Casp3), and four pluripotency/differentiation related genes, Oct4, SRY (sex determining region Y)-box 2 (Sox2), Cdx2, and colony-stimulating factor 1 receptor (Csf1r), were measured by real-time RT-PCR. TSA treatment resulted in the high expression of antiapoptotic gene Bcl-xL and low expression of pro-apoptotic gene Bax compared to untreated NT embryos, fertilized embryos, or parthenotes. Furthermore, mRNA expression of Cdx2 was higher in NT-TSA embryos than in NT and in vitro fertilization (IVF) counterparts. Additionally, low expression of microRNA (mir)-21 in NT embryos was enhanced following TSA treatment. These results suggest that TSA positively regulates nuclear reprogramming, and TSA may increased resistance or reduced signal for induction of apoptosis.
authors.affiliation: Center for Laboratory Animal, Jilin University, 5333 Xi'an Road, Changchun, Jilin Province, People's Republic of China. xscui88@hotmail.com
authors.forename: Xiang-Shun
authors.initials: XS
authors.lastname: Cui
authors.forename: Yong-Nan
authors.initials: YN
authors.lastname: Xu
authors.forename: Xing-Hui
authors.initials: XH
authors.lastname: Shen
authors.forename: Li-Qun
authors.initials: LQ
authors.lastname: Zhang
authors.forename: Jia-Bao
authors.initials: JB
authors.lastname: Zhang
authors.forename: Nam-Hyung
authors.initials: NH
authors.lastname: Kim
mesh_terms: D000818:Animals; D051017:Apoptosis Regulatory Proteins / Q000096:biosynthesis*; D002417:Cattle; D019976:Cloning, Organism; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006877:Hydroxamic Acids / Q000494:pharmacology*; D008297:Male; D035683:MicroRNAs / Q000096:biosynthesis; D053652:Nuclear Transfer Techniques; D010312:Parthenogenesis / Q000187:drug effects; D012333:RNA, Messenger / Q000096:biosynthesis
chemical_list: D051017:Apoptosis Regulatory Proteins; D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; D035683:MicroRNAs; D012333:RNA, Messenger; C012589:trichostatin A