TY - JOUR
T1 - MicroRNA regulation via DNA methylation during the morula to blastocyst transition in mice
AU - Lee, Yee Ming
AU - Chen, Huei Wen
AU - Maurya, Pawan Kumar
AU - Su, Ching Mao
AU - Tzeng, Chii Ruey
N1 - Funding Information:
This work was supported by grants from the National Science Council Grants (96-2628-B-038-008-MY3; 99-2314-B-038-003-MY3).
PY - 2012/4
Y1 - 2012/4
N2 - Epigenetic regulation is responsible for transcriptional silencing of genes and parental imprinting. This study addresses the question whether microRNAs (miRNAs) could be affected by DNA methylation during morula-blastocyst transition. Mouse embryos were treated with/without a DNA methyltransferase inhibitor (5-aza-2′-deoxycytidine, 5-aza-dC, 10 nM-5 μM). Changes of miRNAs were analyzed by quantitative real-time (Q-PCR)-based megaplex pre-amp microRNA assays. Development from morula to blastocyst in mice was inhibited by 5-aza-dC in a dose-dependent manner (10 nM-5 μM), with half of the embryos arrested at morula stage when treated with levels of 5-aza-dC as low as 50 nM. In total, 48 down-regulated microRNAs and 17 up-regulated microRNAs (≥5-fold changes) were identified after 5-aza-dC treatment, including let-7e, mir-20a, mir-21, mir-34b, mir-128b and mir-452. Their predicted targets were selected based on software analysis, published databases and further confirmed by Q-PCR. At least eight targets, including dnmt3a, jagged 1, sp1, edg2, abcg4, numa1, tmsb10 and csf1r were confirmed. In conclusion, 5-aza-dC-modified microRNA profiles and identification of the microRNA's targets during the morula to blastocyst stage in mice provide information that helps us to explore the relationship between fertility, microRNA regulation and epigenetic intervention.
AB - Epigenetic regulation is responsible for transcriptional silencing of genes and parental imprinting. This study addresses the question whether microRNAs (miRNAs) could be affected by DNA methylation during morula-blastocyst transition. Mouse embryos were treated with/without a DNA methyltransferase inhibitor (5-aza-2′-deoxycytidine, 5-aza-dC, 10 nM-5 μM). Changes of miRNAs were analyzed by quantitative real-time (Q-PCR)-based megaplex pre-amp microRNA assays. Development from morula to blastocyst in mice was inhibited by 5-aza-dC in a dose-dependent manner (10 nM-5 μM), with half of the embryos arrested at morula stage when treated with levels of 5-aza-dC as low as 50 nM. In total, 48 down-regulated microRNAs and 17 up-regulated microRNAs (≥5-fold changes) were identified after 5-aza-dC treatment, including let-7e, mir-20a, mir-21, mir-34b, mir-128b and mir-452. Their predicted targets were selected based on software analysis, published databases and further confirmed by Q-PCR. At least eight targets, including dnmt3a, jagged 1, sp1, edg2, abcg4, numa1, tmsb10 and csf1r were confirmed. In conclusion, 5-aza-dC-modified microRNA profiles and identification of the microRNA's targets during the morula to blastocyst stage in mice provide information that helps us to explore the relationship between fertility, microRNA regulation and epigenetic intervention.
KW - Blastocyst
KW - DNA methylation
KW - Epigenetic
KW - MicroRNA
KW - Morula
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U2 - 10.1093/molehr/gar072
DO - 10.1093/molehr/gar072
M3 - Article
C2 - 22053057
AN - SCOPUS:84863378439
SN - 1360-9947
VL - 18
SP - 184
EP - 193
JO - Molecular Human Reproduction
JF - Molecular Human Reproduction
IS - 4
M1 - gar072
ER -