title: Altered gene expression profiles in the brain, kidney, and lung of deceased neonatal cloned pigs.
abstract: Limited studies have been published analyzing the gene expression patterns of cloned pigs. We compared the expression profiles of brain, kidney, and lung tissues, representing each of the three germ layers, of deceased neonatal cloned pigs with those of age-matched controls using a 13K oligonucleotide microarray. We found 42 (0.7% of total genes analyzed), 178 (2.9%), and 121 (1.9%) genes differentially expressed in the brain, kidney, and lung of clones, respectively, when compared with the corresponding organs from controls (fold change >1.5, p < 0.05, false discovery rate (FDR) = 0.05). These expression aberrations could potentially cause the following pathological anomalies in clones: diabetic nephropathy in the kidney and dysregulated surfactant homeostasis in the lung. Interestingly, upregulated expression of genes belonging to the MAPK pathway was observed in all three organs. To investigate whether the differences in levels of gene expression were caused by differential DNA methylation, the global DNA methylation level was measured by high-performance liquid chromatography. In controls, global concentration of methylated cytosine was 5.35%, whereas clones had significantly hypomethylated genomic DNA (4.57%). Bisulfite-pyrosequencing analyses of the promoter regions of differentially expressed candidate genes, c-MYC, Period 1 (PER1), Cathepsin L (CTSL), and Follistatin (FS), however, did not show any differences in the degree of DNA methylation between controls and clones. Our findings demonstrate that deceased neonatal cloned pigs have considerable gene expression abnormalities, which may have contributed to the death of the animals.
authors: Center for Regenerative Biology, Department of Animal Science, University of Connecticut , Storrs, USA.
authors: Joonghoon
authors: J
authors: Park
authors: Sadie L
authors: SL
authors: Marjani
authors: Liangxue
authors: L
authors: Lai
authors: Melissa
authors: M
authors: Samuel
authors: David
authors: D
authors: Wax
authors: Steven R
authors: SR
authors: Davis
authors: Richard S
authors: RS
authors: Bruno
authors: Randall S
authors: RS
authors: Prather
authors: Xiangzhong
authors: X
authors: Yang
authors: Xiuchun Cindy
authors: XC
authors: Tian
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001921:Brain / Q000378:metabolism; D019976:Cloning, Organism / Q000662:veterinary*; D019175:DNA Methylation; D020869:Gene Expression Profiling; D007668:Kidney / Q000378:metabolism; D008168:Lung / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D034421:Sus scrofa / Q000235:genetics* / Q000378:metabolism