Growth regulation, imprinting, and epigenetic transcription-related gene expression differs in lung of deceased transgenic cloned and normal goats Li Meng a,1 , Ruo-Xin Jia a,1 , Yan-Yan Sun b , Zi-Yu Wang a , Yong-Jie Wan a , Yan-Li Zhang a , Bu-Shuai Zhong a , Feng Wang a, * a Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing, PR China b Animal Breeding and Genomics Centre, Wageningen University, Wageningen, The Netherlands article info Article history: Received 5 August 2013 Received in revised form 20 October 2013 Accepted 22 October 2013 Keywords: Growth-regulation Imprinting Epigenetic transcription Transgenic cloned goat qRT-PCR abstract Somatic cell nuclear transfer (SCNT) is a promising technique to produce mammalian transgenic clones. Only a small proportion of manipulated embryos, however, can develop into viable offspring. The abnormal growth and development of cloned animals, further- more, are accompanied by aberrant lung development. Our objective was to investigate molecular background of lung developmental problems in transgenic (random insertion of exogenous DNA) cloned goats. We examined expression of 15 genes involved in growth regulation, imprinting, and epigenetic transcription in lung tissue of deceased transgenic cloned and normal goats of various ages. Compared with normal goats of the same age from conventional reproduction, expression of 13 genes (BMP4, FGF10, GHR, HGFR, PDGFR, RABP , VEGF, H19, CDKN I C, PCAF, MeCP 2 , HDAC1 , and Dnmt3b) decreased in transgenic cloned goats that died at or shortly after birth; Expression of eight genes (FGF10, PDGFR, RABP , VEGF, PCAF, HDAC1 , MeCP 2 , and Dnmt3b) decreased in fetal death of transgenic cloned goats. Expression of two epigenetic transcription genes (PCAF and Dnmt3b) decreased in disease death of transgenic cloned goats (1–4 months old). Disruptions in gene expression might be associated with the high neonatal mortality in transgenic cloned animals. These findings have implications in understanding the low efficiency of transgenic cloning. Ó 2014 Elsevier Inc. All rights reserved. 1. Introduction Somatic cell nuclear transfer (SCNT) [1,2], which uses preselected genetically modified cells as donor nuclei [3], opens new horizons for application of transgenic technol- ogies in livestock animals. The production of the first transgenic cloned goat using SCNT [4] came after reports of the first transgenic cloned sheep, cow, and mouse, and was followed by cloning of many other large animal species. Cloning of other species can reach the efficiency up to 4% (total live kids/embryos transferred), whereas average efficiency in goats is only 2.6% [5]. The low overall success rate is a cumulative result of inefficiencies at each stage of the process, including embryonic, fetal, prenatal, and neonatal loss, and production of abnormal offspring. Initial and subsequent SCNT-derived large animals including goats, furthermore, still have a lot of health issues and a shortened life span [6,7]. The primary reason for low efficiency of cloning and health problems in cloned animals is incomplete reprog- ramming of donor somatic cell nuclei, which might lead to abnormal or even lack of expression of some impor- tant growth regulation, imprinting, and epigenetic transcription-related genes [8–11]. Deviations in expres- sion of these genes, furthermore, are known to lead to phenotypic abnormalities of cloned animals [12,13]. The * Corresponding author. Tel.: þ86 25 84395381; fax: þ86 25 84395314. E-mail address: caeet@njau.edu.cn (F. Wang). 1 These two authors are co-first authors. Contents lists available at ScienceDirect Theriogenology journal homepage: www.theriojournal.com 0093-691X/$ – see front matter Ó 2014 Elsevier Inc. All rights reserved. http://dx.doi.org/10.1016/j.theriogenology.2013.10.023 Theriogenology 81 (2014) 459–466