Developmental Pluripotency of the Nuclei of Neurons in the Cerebral Cortex of Juvenile Mice

Osada, Tomoharu; Tamamaki, Nobuaki; Song, Si Young; Kakazu, Naoki; Yamazaki, Yukiko; Makino, Hidenari; Sasaki, Ayako; Hirayama, Teruyoshi; Hamada, Shin; Nave, Klaus Armin; Yanagimachi, Ryuzo; Yagi, Takeshi

doi:10.1523/jneurosci.1591-05.2005

Cited by 12 publications

(21 citation statements)

References 28 publications

(32 reference statements)

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“…Although our data showed that aneuploid or fragmented chromosomes were more common, translocations in the DNA of the N-NT embryos were also detected. Since we did not detect translocations in the C-NT DNA, such changes may be specific for N-NT embryos, not for NT embryos in general, and are therefore possibly responsible for the observed inefficient developmental rate of the N-NT embryos [Osada et al, 2005].…”

Section: Discussionmentioning

confidence: 58%

“…Donor neurons were collected from B6 transgenic mice that expressed green fluorescence protein (GFP) specifically in pyramidal neurons resulting from the use of a genetic marking system [Osada et al, 2005]. Six ntES cell lines derived from the nuclei of neurons (Cam-1, -2, Nex-1, -2, and Gad67-1, -2) and 1 ntES cell line from cumulus cells (Cum-1) were examined in this study ( table 1 ).…”

Section: Mice and Es Cellsmentioning

confidence: 99%

“…Nuclear transfer experiments were carried out as described previously [Osada et al, 2005]. To obtain the condensed chromatids, cells were collected and fixed 2-3 h after injection of the donor nuclei into enucleated oocytes.…”

Section: Nuclear Transfer and Mitotic Spread Preparationmentioning

confidence: 99%

“…We also used ntES cells derived from N-NT embryos at metaphase for SKY karyotyping [Osada et al, 2005]. SKY analysis of 8 ntES cell lines detected 2 clonal chromosomal abnormalities in Cam-2 and Nex-2 ntES cell lines ( table 1 ).…”

Section: Sky Karyotyping Of the Ntes Cells Derived From N-nt Embryo (mentioning

confidence: 99%

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The Chromosomal Constitution of Postmitotic Neurons, Assessed by Neuronal Nuclear Transfer into Oocytes and in ES Cell Lines Derived from Them

Osada

Kakazu

Watanabe

et al. 2009

Cytogenet Genome Res

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Spectral karyotyping (SKY) was used to assess the chromosomal constitution of embryos generated by nuclear transfer (NT) of neuronal nuclei (N-NT) or cumulus cell nuclei (C-NT) into oocytes and of their embryonic stem cell derivatives (ntES cells). We detected chromosomal changes during the first mitotic cleavage and in the condensed chromatids of NT embryos. We also found clonal translocations in the ntES cells that were derived from NT embryos cloned from neuronal nuclei. The differentiation potentials of the ntES cells showing chromosomal rearrangements were partly restricted. Our findings indicate that balanced or unbalanced chromosomal translocations can occur in early NT embryogenesis, suggesting that a DNA repair system is activated during both NT embryogenesis and ntES cell establishment. We observed a higher incidence of chromosomal changes in N-NT than in C-NT embryos, which may reflect a higher frequency of double-stranded (ds) DNA breaks in the neuronal genome.

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Section: Discussionmentioning

confidence: 58%

Section: Mice and Es Cellsmentioning

confidence: 99%

Section: Nuclear Transfer and Mitotic Spread Preparationmentioning

confidence: 99%

Section: Sky Karyotyping Of the Ntes Cells Derived From N-nt Embryo (mentioning

confidence: 99%

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The Chromosomal Constitution of Postmitotic Neurons, Assessed by Neuronal Nuclear Transfer into Oocytes and in ES Cell Lines Derived from Them

Osada

Kakazu

Watanabe

et al. 2009

Cytogenet Genome Res

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“…Unexpectedly, the best organ for source of donor nuclei was the brain, although the specific cell type (or types) of the donor cells has not been determined. Cloned mice have been obtained using the nuclei of embryonic or newborn neural cells (19)(20)(21). However, to our knowledge, no cloned mice have been generated from adult brain cells, and we have also failed to generate cloned mice from fresh adult brain tissues.…”

Section: Discussionmentioning

confidence: 99%

Production of healthy cloned mice from bodies frozen at −20°C for 16 years

Wakayama¹,

Ohta²,

Hikichi³

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

112

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Cloning animals by nuclear transfer provides an opportunity to preserve endangered mammalian species. However, it has been suggested that the ''resurrection'' of frozen extinct species (such as the woolly mammoth) is impracticable, as no live cells are available, and the genomic material that remains is inevitably degraded. Here we report production of cloned mice from bodies kept frozen at ؊20°C for up to 16 years without any cryoprotection. As all of the cells were ruptured after thawing, we used a modified cloning method and examined nuclei from several organs for use in nuclear transfer attempts. Using brain nuclei as nuclear donors, we established embryonic stem cell lines from the cloned embryos. Healthy cloned mice were then produced from these nuclear transferred embryonic stem cells by serial nuclear transfer. Thus, nuclear transfer techniques could be used to ''resurrect'' animals or maintain valuable genomic stocks from tissues frozen for prolonged periods without any cryopreservation.brain ͉ cloning ͉ cryopreservation ͉ nuclear transfer ͉ reprogramming C urrent animal cloning techniques using nuclear transfer (1) can reconstruct animals from donor cells; clones so produced differ only in their mitochondrial DNA (2). This technology has produced a variety of cloned animals for scientific and commercial purposes. However, it is not known whether this method could be applied to the recovery of animals from frozen extinct species. Although most, if not all, mammalian cells lose their functional integrity if frozen without cryoprotective reagents, it is conceivable that some aspects of genomic information remain intact even in such dead bodies. By contrast, the genomes of spermatozoa are resistant to freezing (3), and even freeze-drying (4) without cryoprotectants. However, it has been difficult to demonstrate this in somatic cells, due to technical difficulties in assessing genomic integrity.One recent paper has reported on the nuclear integrity of dead cells after freeze-thawing by improved techniques (5). This study successfully produced cloned mouse embryos and generated germ line chimerae via nuclear transferred embryonic stem cell (ntES) cells. However, although all donor cells were dead after thawing, the donor cells were frozen in a medium including polyvinylpyrrolidone, which is known to exhibit cryoprotectant effect (6). Freeze-dried cells were also used for these nuclear transfer attempt, and all cells were dead after rehydration (7,8). Although cloned embryos were obtained from those dead cells following nuclear transfer, no cloned offspring were obtained. Moreover, so far all studies of nuclear transfer using ''dead'' material have used isolated single cells frozen in the laboratory using special media. In dead specimens frozen in natural conditions such as permafrost tundra, the cells of tissue will presumably bind strongly to each other and freeze gradually after death due to the large body size. It remains to be shown whether nuclei can be collected from whole bodies frozen without cryopro...

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Brains in metamorphosis: reprogramming cell identity within the central nervous system

Arlotta

Berninger

2014

Current Opinion in Neurobiology

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During embryonic development, uncommitted pluripotent cells undergo progressive epigenetic changes that lock them into a final differentiated state. Can mammalian cells change identity within the living organism? Direct lineage reprogramming of cells has attracted attention as a means to achieve organ regeneration. However, it is unclear whether cells in the CNS are endowed with the plasticity to reprogram. Neurons in particular are considered among the most immutable cell types, able to retain their class-specific traits for the lifespan of the organism. Here we focus on two experimental paradigms, glia-to-neuron and neuron-to-neuron conversion, to consider how lineage reprogramming has challenged the notion of CNS immutability, paving the way for the application of reprogramming strategies to reshape neurons and circuits in vivo.

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Developmental Pluripotency of the Nuclei of Neurons in the Cerebral Cortex of Juvenile Mice

Cited by 12 publications

References 28 publications

The Chromosomal Constitution of Postmitotic Neurons, Assessed by Neuronal Nuclear Transfer into Oocytes and in ES Cell Lines Derived from Them

The Chromosomal Constitution of Postmitotic Neurons, Assessed by Neuronal Nuclear Transfer into Oocytes and in ES Cell Lines Derived from Them

Production of healthy cloned mice from bodies frozen at −20°C for 16 years

Brains in metamorphosis: reprogramming cell identity within the central nervous system

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