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Planarian MBD2/3 is required for adult stem cell pluripotency independently of DNA methylation.

Jaber-Hijazi F, Lo PJ, Mihaylova Y, Foster JM, Benner JS, Tejada Romero B, Chen C, Malla S, Solana J, Ruzov A, Aziz Aboobaker A - Dev. Biol. (2013)

Bottom Line: Here we show the planarian methyl-CpG Binding Domain 2/3 (mbd2/3) gene is required for pASC differentiation during regeneration and tissue homeostasis.The genome does not have detectable levels of 5-methylcytosine (5(m)C) and we find no role for a potential DNA methylase.We conclude that MBD proteins may have had an ancient role in broadly controlling animal stem cell pluripotency, but that DNA methylation is not involved in planarian stem cell differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Zoology, Tinbergen Building, South Parks Road, University of Oxford, Oxford OX1 3PS, United Kingdom.

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mbd2/3(RNAi) Leads to an accumulation of NB.21.11e+ cells and a decline Smed-AGAT-1+ during regeneration. Double FISH on middle pieces shows maintenance of Smed-NB.21.11e+ cells and loss of Smed-AGAT-1+ cells during regeneration in mbd2/3(RNAi) animals (A). Anterior to the left. Scale bar, 200 µm. mbd2/3(RNAi) leads a significant increase in NB.21.11e+ cells and a significant decrease in Smed-AGAT-1+ cells (B), which is quantification of (A); n=3. Standard error bars are shown. ⁎ Indicates p<0.05. Accumulation of NB.21.11e+ cells and a decline in AGAT-1+ cells is apparent throughout regenerating head, middle and tail pieces (C) and (D) Scale bar 500 µm. RT-qPCR analyses of both progeny marker transcripts correlates with the accumulation and loss of early and late stem cell progeny (E). The alternate progeny marker Smed-CYP1A1-1 is also depleted during regeneration by mbd2/3(RNAi) during regeneration (F). Scale bar, 200 µm.
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f0025: mbd2/3(RNAi) Leads to an accumulation of NB.21.11e+ cells and a decline Smed-AGAT-1+ during regeneration. Double FISH on middle pieces shows maintenance of Smed-NB.21.11e+ cells and loss of Smed-AGAT-1+ cells during regeneration in mbd2/3(RNAi) animals (A). Anterior to the left. Scale bar, 200 µm. mbd2/3(RNAi) leads a significant increase in NB.21.11e+ cells and a significant decrease in Smed-AGAT-1+ cells (B), which is quantification of (A); n=3. Standard error bars are shown. ⁎ Indicates p<0.05. Accumulation of NB.21.11e+ cells and a decline in AGAT-1+ cells is apparent throughout regenerating head, middle and tail pieces (C) and (D) Scale bar 500 µm. RT-qPCR analyses of both progeny marker transcripts correlates with the accumulation and loss of early and late stem cell progeny (E). The alternate progeny marker Smed-CYP1A1-1 is also depleted during regeneration by mbd2/3(RNAi) during regeneration (F). Scale bar, 200 µm.

Mentions: We observed that Smed-NB.21.11e+ cells were produced during regeneration in mbd2/3(RNAi) animals, and accumulated in regeneration blastemas when compared to control animals (Fig. 5A–C). However, the later Smed-AGAT-1+ stem cell progeny were depleted in mbd2/3(RNAi) animals during regeneration (Fig. 5A, B and D. These data correlated with increased transcript levels for Smed-NB.21.11e and decreased transcript levels of Smed-AGAT-1 (Fig. 5E). Also during regeneration we observed a reduction in the expression of Smed-CYP1A1-1, another marker of late pASC progeny (Eisenhoffer et al., 2008) (Fig. 5F).


Planarian MBD2/3 is required for adult stem cell pluripotency independently of DNA methylation.

Jaber-Hijazi F, Lo PJ, Mihaylova Y, Foster JM, Benner JS, Tejada Romero B, Chen C, Malla S, Solana J, Ruzov A, Aziz Aboobaker A - Dev. Biol. (2013)

mbd2/3(RNAi) Leads to an accumulation of NB.21.11e+ cells and a decline Smed-AGAT-1+ during regeneration. Double FISH on middle pieces shows maintenance of Smed-NB.21.11e+ cells and loss of Smed-AGAT-1+ cells during regeneration in mbd2/3(RNAi) animals (A). Anterior to the left. Scale bar, 200 µm. mbd2/3(RNAi) leads a significant increase in NB.21.11e+ cells and a significant decrease in Smed-AGAT-1+ cells (B), which is quantification of (A); n=3. Standard error bars are shown. ⁎ Indicates p<0.05. Accumulation of NB.21.11e+ cells and a decline in AGAT-1+ cells is apparent throughout regenerating head, middle and tail pieces (C) and (D) Scale bar 500 µm. RT-qPCR analyses of both progeny marker transcripts correlates with the accumulation and loss of early and late stem cell progeny (E). The alternate progeny marker Smed-CYP1A1-1 is also depleted during regeneration by mbd2/3(RNAi) during regeneration (F). Scale bar, 200 µm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3824064&req=5

f0025: mbd2/3(RNAi) Leads to an accumulation of NB.21.11e+ cells and a decline Smed-AGAT-1+ during regeneration. Double FISH on middle pieces shows maintenance of Smed-NB.21.11e+ cells and loss of Smed-AGAT-1+ cells during regeneration in mbd2/3(RNAi) animals (A). Anterior to the left. Scale bar, 200 µm. mbd2/3(RNAi) leads a significant increase in NB.21.11e+ cells and a significant decrease in Smed-AGAT-1+ cells (B), which is quantification of (A); n=3. Standard error bars are shown. ⁎ Indicates p<0.05. Accumulation of NB.21.11e+ cells and a decline in AGAT-1+ cells is apparent throughout regenerating head, middle and tail pieces (C) and (D) Scale bar 500 µm. RT-qPCR analyses of both progeny marker transcripts correlates with the accumulation and loss of early and late stem cell progeny (E). The alternate progeny marker Smed-CYP1A1-1 is also depleted during regeneration by mbd2/3(RNAi) during regeneration (F). Scale bar, 200 µm.
Mentions: We observed that Smed-NB.21.11e+ cells were produced during regeneration in mbd2/3(RNAi) animals, and accumulated in regeneration blastemas when compared to control animals (Fig. 5A–C). However, the later Smed-AGAT-1+ stem cell progeny were depleted in mbd2/3(RNAi) animals during regeneration (Fig. 5A, B and D. These data correlated with increased transcript levels for Smed-NB.21.11e and decreased transcript levels of Smed-AGAT-1 (Fig. 5E). Also during regeneration we observed a reduction in the expression of Smed-CYP1A1-1, another marker of late pASC progeny (Eisenhoffer et al., 2008) (Fig. 5F).

Bottom Line: Here we show the planarian methyl-CpG Binding Domain 2/3 (mbd2/3) gene is required for pASC differentiation during regeneration and tissue homeostasis.The genome does not have detectable levels of 5-methylcytosine (5(m)C) and we find no role for a potential DNA methylase.We conclude that MBD proteins may have had an ancient role in broadly controlling animal stem cell pluripotency, but that DNA methylation is not involved in planarian stem cell differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Zoology, Tinbergen Building, South Parks Road, University of Oxford, Oxford OX1 3PS, United Kingdom.

Show MeSH