Multi-locus DNA methylation analysis of imprinted genes in cattle from somatic cell nuclear transfer

Multi-locus methylation defects (MLMDs) in imprinted loci have been reported in Beckwith-Wiedemann Syndrome (BWS). Large offspring syndrome (LOS), a phenotypic subgroup of abnormal offspring syndrome (AOS), is considered a molecular and phenotypic model for BWS. Both LOS and BWS have presented epige...

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Published in:	Theriogenology Vol. 186; pp. 95 - 107
Main Authors:	Mangiavacchi, Paula Magnelli, Caldas-Bussiere, Maria Clara, Mendonça, Mariana da Silva, Rumpf, Rodolfo, Lemos Júnior, Paulo Edson Soares, Alves, Carla Soares, Carneiro, Warlei da Silva, Dias, Angelo José Burla, Rios, Álvaro Fabrício Lopes
Format:	Journal Article
Language:	English
Published:	United States Elsevier Inc 01-07-2022
Subjects:	Abnormal offspring syndrome Animals Beckwith-wiedemann Beckwith-Wiedemann Syndrome - genetics Beckwith-Wiedemann Syndrome - veterinary Cattle - genetics Cattle Diseases - genetics DNA Methylation Epigenesis, Genetic Genomic Imprinting Large offspring syndrome Multi-locus imprinting disturbances Multi-locus methylation defects Nuclear Transfer Techniques - veterinary Beckwith-wiedemann Abnormal offspring syndrome Multi-locus methylation defects Large offspring syndrome Multi-locus imprinting disturbances
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Summary:	Multi-locus methylation defects (MLMDs) in imprinted loci have been reported in Beckwith-Wiedemann Syndrome (BWS). Large offspring syndrome (LOS), a phenotypic subgroup of abnormal offspring syndrome (AOS), is considered a molecular and phenotypic model for BWS. Both LOS and BWS have presented epigenetic defects in some common imprinted loci. In this study, methylation-specific restriction digestion assay - quantitative PCR was used to analyze the DNA methylation pattern in differentially methylated regions (DMRs) of the H19 (H19-DMR), KCNQ1OT1 (KvDMR1) and PEG1/MEST (PEG1-DMR) genes in bovine clone tissues from calves that did not survive after birth. Individual and tissue-specific changes in DNA methylation levels in the bovine KvDMR1, H19-DMR, and PEG1-DMR were observed. In contrast to what has been reported in the literature on BWS and AOS/LOS, the KvDMR1 showed gain (GOM) and loss (LOM) of DNA methylation. LOM and GOM events were found in the DMRs studied in animals produced by the same nucleus donor cell line. This is the first report of epimutations in the PEG1-DMR and GOM at the KvDMR1 found in bovine clones. The findings showed that epigenetic modification in imprinted loci in cloned cattle occurred in a multi-locus pattern similar to that seen in human imprinting disorders. Other multi-locus analyzes must be done to elucidate the MLMD pattern in AOS in bovine clones. •Bovine clones that did not survive after births with LOS/AOS characteristics.•This is the first study to detect GOM pattern in KvDMR1 region in AOS clones.•MLMD event was observed in a tissue-specific manner in bovine clones.•5 mC% difference in clones from the same nucleus donor occurred to errors in epigenetics reprogramming.•The PEG1-DMR region showed a significant increase in DNA methylation in AOS clones.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0093-691X 1879-3231
DOI:	10.1016/j.theriogenology.2022.04.006