Abstract
Imprinting disorders induced by somatic cell nuclear transfer (SCNT) usually lead to the abnormalities of cloned animals and low cloning efficiency. Histone deacetylase inhibitors have been shown to improve gene expression, genomic methylation reprogramming and the development of cloned embryos, however, the imprinting statuses in these treated embryos and during their subsequent development remain poorly studied. In this study, we investigated the dynamics of H19/Igf2 methylation and transcription in porcine cloned embryos treated with trichostatin A (TSA), and examined H19/Igf2 imprinting patterns in cloned fetuses and piglets. Our results showed that compared with the maintenance of H19/Igf2 methylation in fertilized embryos, cloned embryos displayed aberrant H19/Igf2 methylation and lower H19/Igf2 transcripts. When TSA enhanced the development of cloned embryos, the disrupted H19/Igf2 imprinting was largely rescued in these treated embryos, more similar to those detected in fertilized counterparts. Further studies displayed that TSA effectively rescued the disrupted imprinting of H19/Igf2 in cloned fetuses and piglets, prevented the occurrence of cloned fetus and piglet abnormalities, and enhanced the full-term development of cloned embryos. In conclusion, our results demonstrated that aberrant imprinting induced by SCNT led to the abnormalities of cloned fetuses and piglets and low cloning efficiency, and TSA rescued the disrupted imprinting in cloned embryos, fetuses and piglets, and prevented the occurrence of cloned fetus and piglet abnormalities, thereby improving the development of cloned embryos. This study would have important implications in improving cloning efficiency and the health of cloned animals.
Highlights
Though somatic cell nuclear transfer (SCNT) has been achieved in many species, overall cloning efficiency is still low, and the developmental abnormalities, including low birth rates, placental defect and large offspring syndrome, etc., frequently occur [1,2,3], limiting the application of SCNT technology in basic research, agriculture and medicine.PLOS ONE | DOI:10.1371/journal.pone.0126607 May 11, 2015trichostatin A (TSA) Rescues the Disrupted Imprinting role in study design, data collection and analysis, decision to publish, or preparation of the manuscript
We investigated the dynamics of H19/Igf2 methylation and transcription in porcine cloned embryos treated with trichostatin A (TSA), and examined H19/Igf2 imprinting patterns in cloned fetuses and piglets
Our results demonstrated that aberrant imprinting induced by SCNT led to the abnormalities of cloned fetuses and piglets and low cloning efficiency, and TSA rescued the disrupted imprinting in cloned embryos, fetuses and piglets, and prevented the occurrence of cloned fetus and piglet abnormalities, thereby improving the development of cloned embryos
Summary
Though somatic cell nuclear transfer (SCNT) has been achieved in many species, overall cloning efficiency is still low, and the developmental abnormalities, including low birth rates, placental defect and large offspring syndrome, etc., frequently occur [1,2,3], limiting the application of SCNT technology in basic research, agriculture and medicine.PLOS ONE | DOI:10.1371/journal.pone.0126607 May 11, 2015TSA Rescues the Disrupted Imprinting role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Though somatic cell nuclear transfer (SCNT) has been achieved in many species, overall cloning efficiency is still low, and the developmental abnormalities, including low birth rates, placental defect and large offspring syndrome, etc., frequently occur [1,2,3], limiting the application of SCNT technology in basic research, agriculture and medicine. TSA Rescues the Disrupted Imprinting role in study design, data collection and analysis, decision to publish, or preparation of the manuscript
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