Platypus Induced Pluripotent Stem Cells: the Unique Pluripotency Signature of a Monotreme

Deanne Jeanette Whitworth, Jason Ionnis Limnios, Marie-Emilie Gauthier, Prasanna Weeratunga, Dmitry A Ovchinnikov, Gregory Baillie, Sean M Grimmond, Jennifer A M Graves, Ernst Jurgen Wolvetang

Research output: Contribution to journalArticleResearchpeer-review

Abstract

The platypus (Ornithorhynchus anatinus) is an egg-laying monotreme mammal whose ancestors diverged approximately 166 million years ago from the evolutionary pathway that eventually gave rise to both marsupial and eutherian mammals. Consequently, its genome is an extraordinary amalgam of both ancestral reptilian and derived mammalian features. To gain insight into the evolution of mammalian pluripotency we have generated induced pluripotent stem cells (piPSCs) from the platypus. Deep sequencing of the piPSC transcriptome revealed that piPSCs robustly express the core eutherian pluripotency factors OCT4, SOX2 and NANOG. Given the more extensive role of SOX3 over SOX2 in avian pluripotency, our data indicate that between 315 million years and 166 million years ago primitive mammals replaced the role of SOX3 in the vertebrate pluripotency network with SOX2. DAX1/NR0B1 is not expressed in piPSCs and an analysis of the platypus DAX1 promoter revealed the absence of a proximal SOX2-binding DNA motif known to be critical for DAX1 expression in eutherian pluripotent stem cells, suggesting that the acquisition of SOX2 responsiveness by DAX1 has facilitated its recruitment into the pluripotency network of eutherians. Using the RNAseq data, we were also able to demonstrate that in both fibroblasts and piPSCs the expression ratio of X chromosomes to autosomes (X1-5 X1-5:AA) is approximately equal to 1 indicating that there is no upregulation of X-linked genes. Finally, the RNAseq data also allowed us to explore the process of X-linked gene inactivation in the platypus, where we determined that for any given gene there is no preference for silencing of the maternal or paternal allele; that is, within a population of cells the silencing of X-linked genes is not imprinted.

LanguageEnglish
Pages151-164
Number of pages14
JournalStem Cells and Development
Volume28
Issue number3
Early online date9 Jan 2019
DOIs
Publication statusPublished - 1 Feb 2019

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Platypus
monotreme
Induced Pluripotent Stem Cells
Pluripotent Stem Cells
X-Linked Genes
stem cells
stem
Mammals
mammals
mammal
gene
genes
High-Throughput Nucleotide Sequencing
Marsupialia
Nucleotide Motifs
DNA-binding domains
autosomes
X Chromosome
Gene Silencing
X chromosome

Cite this

Whitworth, D. J., Limnios, J. I., Gauthier, M-E., Weeratunga, P., Ovchinnikov, D. A., Baillie, G., ... Wolvetang, E. J. (2019). Platypus Induced Pluripotent Stem Cells: the Unique Pluripotency Signature of a Monotreme. Stem Cells and Development, 28(3), 151-164. https://doi.org/10.1089/scd.2018.0179
Whitworth, Deanne Jeanette ; Limnios, Jason Ionnis ; Gauthier, Marie-Emilie ; Weeratunga, Prasanna ; Ovchinnikov, Dmitry A ; Baillie, Gregory ; Grimmond, Sean M ; Graves, Jennifer A M ; Wolvetang, Ernst Jurgen. / Platypus Induced Pluripotent Stem Cells : the Unique Pluripotency Signature of a Monotreme. In: Stem Cells and Development. 2019 ; Vol. 28, No. 3. pp. 151-164.
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abstract = "The platypus (Ornithorhynchus anatinus) is an egg-laying monotreme mammal whose ancestors diverged approximately 166 million years ago from the evolutionary pathway that eventually gave rise to both marsupial and eutherian mammals. Consequently, its genome is an extraordinary amalgam of both ancestral reptilian and derived mammalian features. To gain insight into the evolution of mammalian pluripotency we have generated induced pluripotent stem cells (piPSCs) from the platypus. Deep sequencing of the piPSC transcriptome revealed that piPSCs robustly express the core eutherian pluripotency factors OCT4, SOX2 and NANOG. Given the more extensive role of SOX3 over SOX2 in avian pluripotency, our data indicate that between 315 million years and 166 million years ago primitive mammals replaced the role of SOX3 in the vertebrate pluripotency network with SOX2. DAX1/NR0B1 is not expressed in piPSCs and an analysis of the platypus DAX1 promoter revealed the absence of a proximal SOX2-binding DNA motif known to be critical for DAX1 expression in eutherian pluripotent stem cells, suggesting that the acquisition of SOX2 responsiveness by DAX1 has facilitated its recruitment into the pluripotency network of eutherians. Using the RNAseq data, we were also able to demonstrate that in both fibroblasts and piPSCs the expression ratio of X chromosomes to autosomes (X1-5 X1-5:AA) is approximately equal to 1 indicating that there is no upregulation of X-linked genes. Finally, the RNAseq data also allowed us to explore the process of X-linked gene inactivation in the platypus, where we determined that for any given gene there is no preference for silencing of the maternal or paternal allele; that is, within a population of cells the silencing of X-linked genes is not imprinted.",
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Whitworth, DJ, Limnios, JI, Gauthier, M-E, Weeratunga, P, Ovchinnikov, DA, Baillie, G, Grimmond, SM, Graves, JAM & Wolvetang, EJ 2019, 'Platypus Induced Pluripotent Stem Cells: the Unique Pluripotency Signature of a Monotreme' Stem Cells and Development, vol. 28, no. 3, pp. 151-164. https://doi.org/10.1089/scd.2018.0179

Platypus Induced Pluripotent Stem Cells : the Unique Pluripotency Signature of a Monotreme. / Whitworth, Deanne Jeanette; Limnios, Jason Ionnis; Gauthier, Marie-Emilie; Weeratunga, Prasanna; Ovchinnikov, Dmitry A; Baillie, Gregory; Grimmond, Sean M; Graves, Jennifer A M; Wolvetang, Ernst Jurgen.

In: Stem Cells and Development, Vol. 28, No. 3, 01.02.2019, p. 151-164.

Research output: Contribution to journalArticleResearchpeer-review

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