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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 homeostasis. Smed-NB.21.11e expressing early progeny cells during homeostasis are maintained and they accumulate at sites of anterior regression (black arrow) in mbd2/3(RNAi) animals (A). Reduction in the number of Smed-AGAT-1 expression late pASC progeny cells during tissue homeostasis in mbd2/3(RNAi) animals, with loss in an anterior to posterior direction (B). Scale bars, 500 µm. (C) Double FISH on intact animals during homeostasis to visualize neoblast progeny cell formation. +: eyes. Scale bar, 200 µm. Quantification of (C) with n=3. ⁎⁎ Indicates p<0.01 as determined by Student's t-test. (D) The alternate early progeny marker Smed-NB.32.1g is also maintained while the alternate late progeny marker Smed-CYP1A1-1 is depleted (E). Scale bars, 500 µm.
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f0030: mbd2/3(RNAi) Leads to an accumulation of NB.21.11e+ cells and a decline Smed-AGAT-1+ during homeostasis. Smed-NB.21.11e expressing early progeny cells during homeostasis are maintained and they accumulate at sites of anterior regression (black arrow) in mbd2/3(RNAi) animals (A). Reduction in the number of Smed-AGAT-1 expression late pASC progeny cells during tissue homeostasis in mbd2/3(RNAi) animals, with loss in an anterior to posterior direction (B). Scale bars, 500 µm. (C) Double FISH on intact animals during homeostasis to visualize neoblast progeny cell formation. +: eyes. Scale bar, 200 µm. Quantification of (C) with n=3. ⁎⁎ Indicates p<0.01 as determined by Student's t-test. (D) The alternate early progeny marker Smed-NB.32.1g is also maintained while the alternate late progeny marker Smed-CYP1A1-1 is depleted (E). Scale bars, 500 µm.

Mentions: During tissue homeostasis in intact animals Smed-NB.21.11e+ cells were maintained in mbd2/3(RNAi) animals and accumulated at the sites of anterior regression (Fig. 6A, C and D). On the other hand, later Smed-AGAT-1+ stem cell progeny were depleted in mbd2/3(RNAi) animals (Fig. 6B–D). This resulted in animals with accumulated Smed-NB.21.11e+ cells and depleted Smed-AGAT-1+ cells by 3 weeks of homeostasis (Fig. 6C). The data observed for Smed-NB.21.11e+ and Smed-AGAT-1+ cells was supported by an increase in Smed-NB.21.11e transcript levels and a decrease in Smed-AGAT-1 transcript levels (Supplementary Fig. 4A and B). An alternate early progeny marker Smed-NB.32.1g (Eisenhoffer et al., 2008) was also maintained at the third week of homeostasis following mbd2/3(RNAi) (Fig. 6E). The loss of late neoblast progeny in mbd2/3(RNAi) animals during homeostasis was confirmed using the Smed-CYP1A1 marker gene (Fig. 6F). These data suggest that pASCs lacking mbd2/3 are able to give rise to early stem cell progeny, but that these progeny are unable to differentiate correctly, with later stem cell progeny failing to form.


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 homeostasis. Smed-NB.21.11e expressing early progeny cells during homeostasis are maintained and they accumulate at sites of anterior regression (black arrow) in mbd2/3(RNAi) animals (A). Reduction in the number of Smed-AGAT-1 expression late pASC progeny cells during tissue homeostasis in mbd2/3(RNAi) animals, with loss in an anterior to posterior direction (B). Scale bars, 500 µm. (C) Double FISH on intact animals during homeostasis to visualize neoblast progeny cell formation. +: eyes. Scale bar, 200 µm. Quantification of (C) with n=3. ⁎⁎ Indicates p<0.01 as determined by Student's t-test. (D) The alternate early progeny marker Smed-NB.32.1g is also maintained while the alternate late progeny marker Smed-CYP1A1-1 is depleted (E). Scale bars, 500 µm.
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Related In: Results  -  Collection

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

f0030: mbd2/3(RNAi) Leads to an accumulation of NB.21.11e+ cells and a decline Smed-AGAT-1+ during homeostasis. Smed-NB.21.11e expressing early progeny cells during homeostasis are maintained and they accumulate at sites of anterior regression (black arrow) in mbd2/3(RNAi) animals (A). Reduction in the number of Smed-AGAT-1 expression late pASC progeny cells during tissue homeostasis in mbd2/3(RNAi) animals, with loss in an anterior to posterior direction (B). Scale bars, 500 µm. (C) Double FISH on intact animals during homeostasis to visualize neoblast progeny cell formation. +: eyes. Scale bar, 200 µm. Quantification of (C) with n=3. ⁎⁎ Indicates p<0.01 as determined by Student's t-test. (D) The alternate early progeny marker Smed-NB.32.1g is also maintained while the alternate late progeny marker Smed-CYP1A1-1 is depleted (E). Scale bars, 500 µm.
Mentions: During tissue homeostasis in intact animals Smed-NB.21.11e+ cells were maintained in mbd2/3(RNAi) animals and accumulated at the sites of anterior regression (Fig. 6A, C and D). On the other hand, later Smed-AGAT-1+ stem cell progeny were depleted in mbd2/3(RNAi) animals (Fig. 6B–D). This resulted in animals with accumulated Smed-NB.21.11e+ cells and depleted Smed-AGAT-1+ cells by 3 weeks of homeostasis (Fig. 6C). The data observed for Smed-NB.21.11e+ and Smed-AGAT-1+ cells was supported by an increase in Smed-NB.21.11e transcript levels and a decrease in Smed-AGAT-1 transcript levels (Supplementary Fig. 4A and B). An alternate early progeny marker Smed-NB.32.1g (Eisenhoffer et al., 2008) was also maintained at the third week of homeostasis following mbd2/3(RNAi) (Fig. 6E). The loss of late neoblast progeny in mbd2/3(RNAi) animals during homeostasis was confirmed using the Smed-CYP1A1 marker gene (Fig. 6F). These data suggest that pASCs lacking mbd2/3 are able to give rise to early stem cell progeny, but that these progeny are unable to differentiate correctly, with later stem cell progeny failing to form.

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
Related in: MedlinePlus