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Gene activation-associated long noncoding RNAs function in mouse preimplantation development.

Hamazaki N, Uesaka M, Nakashima K, Agata K, Imamura T - Development (2015)

Bottom Line: Expression of these bidirectional promoter-associated noncoding RNAs (pancRNAs) was strongly associated with the upregulation of their cognate genes.Conversely, knockdown of three abundant pancRNAs led to reduced mRNA expression, accompanied by sustained DNA methylation even in the presence of enzymes responsible for DNA demethylation.Thus, this novel class of lncRNAs can modulate the transcription machinery in cis to activate zygotic genes and is important for preimplantation development.

View Article: PubMed Central - PubMed

Affiliation: Department of Biophysics and Global COE Program, Graduate School of Science, Kyoto University, Kitashirakawa-Oiwake, Sakyo-ku, Kyoto 606-8502, Japan Division of Basic Stem Cell Biology, Department of Stem Cell Biology and Medicine, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.

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Effect of pancRNA knockdown on the expression of the counterpart gene during early development. (A) 5′-regions of mouse Il17d, Mospd3 and Tbc1d22a. Vertical lines mark the locations of CpG dinucleotides. Thick horizontal lines denote the regions analyzed by bisulfite sequencing. Primer positions are numbered relative to the TSS of each gene. (B) qPCR analysis of pancRNA and mRNA at the Il17d, Mospd3 and Tbc1d22a loci in MII oocytes and in fertilized 1-cell, 2-cell and 4-cell embryos. Error bars indicate s.e.m. (C) DNA methylation levels of the promoter regions of Il17d, Mospd3 and Tbc1d22a in MII oocytes, sperm and fertilized 1-cell and 2-cell embryos. Filled and open circles indicate methylated and unmethylated cytosines, respectively. *P<0.05; **P<0.01; ***P<0.001.
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DEV116996F2: Effect of pancRNA knockdown on the expression of the counterpart gene during early development. (A) 5′-regions of mouse Il17d, Mospd3 and Tbc1d22a. Vertical lines mark the locations of CpG dinucleotides. Thick horizontal lines denote the regions analyzed by bisulfite sequencing. Primer positions are numbered relative to the TSS of each gene. (B) qPCR analysis of pancRNA and mRNA at the Il17d, Mospd3 and Tbc1d22a loci in MII oocytes and in fertilized 1-cell, 2-cell and 4-cell embryos. Error bars indicate s.e.m. (C) DNA methylation levels of the promoter regions of Il17d, Mospd3 and Tbc1d22a in MII oocytes, sperm and fertilized 1-cell and 2-cell embryos. Filled and open circles indicate methylated and unmethylated cytosines, respectively. *P<0.05; **P<0.01; ***P<0.001.

Mentions: To examine the function of the ZGA-associated pancRNA, we selected highly expressed pancRNAs that were upregulated at the 2-cell stage and whose expression was maintained at a high level in ESCs (supplementary material Table S3), and characterized the three most highly expressed in ESCs, namely those partnered with Il17d (pancIl17d), Mospd3 (pancMospd3) and Tbc1d22a (pancTbc1d22a) (Fig. 2A). First, we examined their expression patterns in the MII oocyte, sperm, and fertilized 1-cell and 2-cell embryo by RT-qPCR (Fig. 2B). We confirmed that all of these pancRNAs were expressed at the 2-cell stage, and found that the expression of Mospd3 and Tbc1d22a mRNAs was also upregulated at the 2-cell stage (Fig. 2B, middle and right panels), whereas the expression of Il17d mRNA was first detected at the 4-cell stage (Fig. 2B, left panel). Thus, expression of the pancRNA preceded or occurred simultaneously with that of the mRNA at these loci during early embryogenesis.Fig. 2.


Gene activation-associated long noncoding RNAs function in mouse preimplantation development.

Hamazaki N, Uesaka M, Nakashima K, Agata K, Imamura T - Development (2015)

Effect of pancRNA knockdown on the expression of the counterpart gene during early development. (A) 5′-regions of mouse Il17d, Mospd3 and Tbc1d22a. Vertical lines mark the locations of CpG dinucleotides. Thick horizontal lines denote the regions analyzed by bisulfite sequencing. Primer positions are numbered relative to the TSS of each gene. (B) qPCR analysis of pancRNA and mRNA at the Il17d, Mospd3 and Tbc1d22a loci in MII oocytes and in fertilized 1-cell, 2-cell and 4-cell embryos. Error bars indicate s.e.m. (C) DNA methylation levels of the promoter regions of Il17d, Mospd3 and Tbc1d22a in MII oocytes, sperm and fertilized 1-cell and 2-cell embryos. Filled and open circles indicate methylated and unmethylated cytosines, respectively. *P<0.05; **P<0.01; ***P<0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4352986&req=5

DEV116996F2: Effect of pancRNA knockdown on the expression of the counterpart gene during early development. (A) 5′-regions of mouse Il17d, Mospd3 and Tbc1d22a. Vertical lines mark the locations of CpG dinucleotides. Thick horizontal lines denote the regions analyzed by bisulfite sequencing. Primer positions are numbered relative to the TSS of each gene. (B) qPCR analysis of pancRNA and mRNA at the Il17d, Mospd3 and Tbc1d22a loci in MII oocytes and in fertilized 1-cell, 2-cell and 4-cell embryos. Error bars indicate s.e.m. (C) DNA methylation levels of the promoter regions of Il17d, Mospd3 and Tbc1d22a in MII oocytes, sperm and fertilized 1-cell and 2-cell embryos. Filled and open circles indicate methylated and unmethylated cytosines, respectively. *P<0.05; **P<0.01; ***P<0.001.
Mentions: To examine the function of the ZGA-associated pancRNA, we selected highly expressed pancRNAs that were upregulated at the 2-cell stage and whose expression was maintained at a high level in ESCs (supplementary material Table S3), and characterized the three most highly expressed in ESCs, namely those partnered with Il17d (pancIl17d), Mospd3 (pancMospd3) and Tbc1d22a (pancTbc1d22a) (Fig. 2A). First, we examined their expression patterns in the MII oocyte, sperm, and fertilized 1-cell and 2-cell embryo by RT-qPCR (Fig. 2B). We confirmed that all of these pancRNAs were expressed at the 2-cell stage, and found that the expression of Mospd3 and Tbc1d22a mRNAs was also upregulated at the 2-cell stage (Fig. 2B, middle and right panels), whereas the expression of Il17d mRNA was first detected at the 4-cell stage (Fig. 2B, left panel). Thus, expression of the pancRNA preceded or occurred simultaneously with that of the mRNA at these loci during early embryogenesis.Fig. 2.

Bottom Line: Expression of these bidirectional promoter-associated noncoding RNAs (pancRNAs) was strongly associated with the upregulation of their cognate genes.Conversely, knockdown of three abundant pancRNAs led to reduced mRNA expression, accompanied by sustained DNA methylation even in the presence of enzymes responsible for DNA demethylation.Thus, this novel class of lncRNAs can modulate the transcription machinery in cis to activate zygotic genes and is important for preimplantation development.

View Article: PubMed Central - PubMed

Affiliation: Department of Biophysics and Global COE Program, Graduate School of Science, Kyoto University, Kitashirakawa-Oiwake, Sakyo-ku, Kyoto 606-8502, Japan Division of Basic Stem Cell Biology, Department of Stem Cell Biology and Medicine, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.

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