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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) Does not affect pASC maintenance or proliferation. (A) The distribution of pASCs assessed by Smedwi-2 expression found to be consistent between mbd2/3(RNAi) animals and their controls during homeostasis even in animals with regressed anterior regions. Smed-H2B expressing cells are also maintained in regenerating and animals undergoing normal homeostasis (B) and (C). Numbers of Smed-H2B expressing cells are not significantly different from controls in these animals (D). Numbers of mitotic cells are not significantly different from controls during regeneration or homeostasis, except at 5 days of regeneration (E) and (F). Scale bars 500 µm, accept in (C) and (D) 100 µm. ⁎ Indicates p<0.05, determined by Student's t-test. Standard error bars are shown.
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f0020: mbd2/3(RNAi) Does not affect pASC maintenance or proliferation. (A) The distribution of pASCs assessed by Smedwi-2 expression found to be consistent between mbd2/3(RNAi) animals and their controls during homeostasis even in animals with regressed anterior regions. Smed-H2B expressing cells are also maintained in regenerating and animals undergoing normal homeostasis (B) and (C). Numbers of Smed-H2B expressing cells are not significantly different from controls in these animals (D). Numbers of mitotic cells are not significantly different from controls during regeneration or homeostasis, except at 5 days of regeneration (E) and (F). Scale bars 500 µm, accept in (C) and (D) 100 µm. ⁎ Indicates p<0.05, determined by Student's t-test. Standard error bars are shown.

Mentions: We observed no effect on the distribution, proliferation or relative numbers of pASCs during tissue homeostasis or regeneration in mbd2/3(RNAi) animals relative to their controls (Fig. 4A–F; Supplementary Fig. 3A–C). Even animals that had regressed to tail stumps after 3 weeks of homeostasis had normal Smedwi-2 expression in pASCs and normal levels of mitosis (Fig. 4A and Supplementary Fig. 3A). We subjected mbd2/3(RNAi) animals to a further round of standard RNAi injections and regeneration and also observed no defect in proliferation (Supplementary Fig. 3C).


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) Does not affect pASC maintenance or proliferation. (A) The distribution of pASCs assessed by Smedwi-2 expression found to be consistent between mbd2/3(RNAi) animals and their controls during homeostasis even in animals with regressed anterior regions. Smed-H2B expressing cells are also maintained in regenerating and animals undergoing normal homeostasis (B) and (C). Numbers of Smed-H2B expressing cells are not significantly different from controls in these animals (D). Numbers of mitotic cells are not significantly different from controls during regeneration or homeostasis, except at 5 days of regeneration (E) and (F). Scale bars 500 µm, accept in (C) and (D) 100 µm. ⁎ Indicates p<0.05, determined by Student's t-test. Standard error bars are shown.
© Copyright Policy
Related In: Results  -  Collection

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

f0020: mbd2/3(RNAi) Does not affect pASC maintenance or proliferation. (A) The distribution of pASCs assessed by Smedwi-2 expression found to be consistent between mbd2/3(RNAi) animals and their controls during homeostasis even in animals with regressed anterior regions. Smed-H2B expressing cells are also maintained in regenerating and animals undergoing normal homeostasis (B) and (C). Numbers of Smed-H2B expressing cells are not significantly different from controls in these animals (D). Numbers of mitotic cells are not significantly different from controls during regeneration or homeostasis, except at 5 days of regeneration (E) and (F). Scale bars 500 µm, accept in (C) and (D) 100 µm. ⁎ Indicates p<0.05, determined by Student's t-test. Standard error bars are shown.
Mentions: We observed no effect on the distribution, proliferation or relative numbers of pASCs during tissue homeostasis or regeneration in mbd2/3(RNAi) animals relative to their controls (Fig. 4A–F; Supplementary Fig. 3A–C). Even animals that had regressed to tail stumps after 3 weeks of homeostasis had normal Smedwi-2 expression in pASCs and normal levels of mitosis (Fig. 4A and Supplementary Fig. 3A). We subjected mbd2/3(RNAi) animals to a further round of standard RNAi injections and regeneration and also observed no defect in proliferation (Supplementary Fig. 3C).

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