Adult murine neurons: their chromatin and chromosome changes and failure to support embryonic development as revealed by nuclear transfer

Osada, Tomoharu; Kusakabe, Hirokazu; Akutsu, Hidenori; Yagi, Takeshi; Yanagimachi, Ryuzo

doi:10.1159/000064037

Cited by 40 publications

(43 citation statements)

References 38 publications

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“…transfer of cortical neuronal nuclei into oocytes indicated that 64% of nuclei contained deviations from a euploid karyotype (16). Despite these considerations, the rate of sex chromosome aneuploidy documented here, Ϸ0.2% for combined X or Y hyperploidy, is substantially greater than other neurobiologically important phenomena, such as adult neurogenesis, where Sphase cells have a prevalence of Ϸ0.03% to 0.05% (17,18).…”

Section: Discussionmentioning

confidence: 53%

Aneuploid neurons are functionally active and integrated into brain circuitry

Kingsbury

Friedman

McConnell

et al. 2005

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

156

130

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The existence of aneuploid cells within the mammalian brain has suggested the influence of genetic mosaicism on normal neural circuitry. However, aneuploid cells might instead be glia, nonneural, or dying cells, which are irrelevant to direct neuronal signaling. Combining retrograde labeling with FISH for chromosome-specific loci, distantly labeled aneuploid neurons were observed in expected anatomical projection areas. Coincident labeling for immediate early gene expression indicated that these aneuploid neurons were functionally active. These results demonstrate that functioning neurons with aneuploid genomes form genetically mosaic neural circuitries as part of the normal organization of the mammalian brain.cerebral cortex ͉ chromosome paint ͉ mosacism A neuploidy, the loss and͞or gain of chromosomes producing numerical deviation from haploid multiples, has documented effects on cellular physiology, particularly in pathophysiological settings such as cancer and Down's syndrome (1, 2). Whereas even the quantitative change of a single-gene product can produce major changes in cellular signaling (3), the effects associated with loss or gain of Ϸ1,000 gene copies on an average chromosome should have even more pronounced changes on a cell's physiology. Approximately 33% of neural progenitor cells display genetic variability, manifested as chromosomal aneuploidy that encompasses both loss and gain of chromosomes (4-7). In the mature brain, aneuploid cells that express neuronal markers have been observed (4, 6). However, an unanswered question is whether these neurons represent functional rather than dying cells, with death being a common fate for aneuploid cells in other systems (8,9). Toward determining their relevance for adult brain function, we examined aneuploid cells in the normal brain for anatomical connectivity and functional activity. MethodsProcedures involving live animals were conducted at the University of California at San Diego (UCSD), approved by the Animal Subjects Committee at UCSD, and conform to National Institutes of Health guidelines and public law.Activation Paradigm for Immediate Early Gene (IEG) Induction. All animals were transferred from the home cage to a clean cage 1 h before killing. To maximize the activation of cortical cells, two mice received additional stimulation that included exposure to a novel male (10) and to various odors (11). Specifically, a novel male was placed in the same cage as the subject male, and the males were allowed to interact for 1-3 min. After the removal of the novel male, subject males were then exposed sequentially to peppermint and banana odors for 15 min each while remaining in the clean cage. Animals were killed 1 h after the beginning of the exposure paradigm, perfused with 0.9% saline, followed by 4% paraformaldehyde in 0.1 M PBS (pH 7.4), and brain tissue was processed as described below.Retrograde Tracer Injections. Ten male 8-week-old mice from three different strains (BalbC͞CR, Swiss Webster, and c129SvJ) received tracer injections. Mice were ane...

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Section: Discussionmentioning

confidence: 53%

Aneuploid neurons are functionally active and integrated into brain circuitry

Kingsbury

Friedman

McConnell

et al. 2005

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

156

130

View full text Add to dashboard Cite

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“…While these data suggest support for the differentiation-hypothesis, numbers were too small to be significant (Yamazaki et al, 2001). Initial attempts to clone viable offspring from fully differentiated cerebral cortex neurons have repeatedly failed (Wakayama et al, 1998;Osada et al, 2002). Until now, no more than a couple of normal looking midgestation fetuses have been cloned from adult cortical neurons using conventional NT procedures, though not for want of trying by, among others, the same highly experienced laboratory that successfully cloned neural progenitors and immature neurons (Wakayama et al, 1998;Makino et al, 2005).…”

Section: Somatic Cellsmentioning

confidence: 96%

Donor cell differentiation, reprogramming, and cloning efficiency: Elusive or illusive correlation?

Oback

Wells

2006

Molecular Reproduction Devel

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Compared to other assisted reproductive technologies, mammalian nuclear transfer (NT) cloning is inefficient in generating viable offspring. It has been postulated that nuclear reprogramming and cloning efficiency can be increased by choosing less differentiated cell types as nuclear donors. This hypothesis is mainly supported by comparative mouse cloning experiments using early blastomeres, embryonic stem (ES) cells, and terminally differentiated somatic donor cells. We have re-evaluated these comparisons, taking into account different NT procedures, the use of donor cells from different genetic backgrounds, sex, cell cycle stages, and the lack of robust statistical significance when post-blastocyst development is compared. We argue that while the reprogrammability of early blastomeres appears to be much higher than that of somatic cells, it has so far not been conclusively determined whether differentiation status affects cloning efficiency within somatic donor cell lineages. Mol. Reprod. Dev. 74: 646-654, 2007. ß

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“…Although several cloned adult mice have been obtained using the nuclei of embryonic, differentiated neural cells (Yamazaki et al, 2001), no live offspring have been cloned from the cerebral cortical neural cells of newborn animals by direct nuclear transfer to enucleated oocytes (Makino et al, 2005). To date, no live offspring have been produced from the cerebral cortical neurons of adult mice (Wakayama et al, 1998;Osada et al, 2002). Here, we report the production of fertile offspring from the cerebral cortical neurons of juvenile mice by first converting the neurons into embryonic stem (ES) cells, then injecting their nuclei into the cytoplasm of enucleated oocytes or transferring the ES cells into the blastocoeles of tetraploid embryos.…”

Section: Introductionmentioning

confidence: 99%

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

Osada¹,

Tamamaki²,

Song³

et al. 2005

J. Neurosci.

Self Cite

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Nuclei isolated from green fluorescent protein-marked neurons in the cerebral cortex of juvenile mice (14 -21 d after birth) were injected into enucleated oocytes that were allowed to develop into blastocysts. Embryonic stem (ES) cell lines were established from the inner cell mass of 76 cloned blastocysts after injecting 2026 neuronal nuclei. Some ES cells were injected individually into enucleated oocytes (nuclear transfer). Other ES cells were transferred into the blastocoeles of tetraploid blastocysts (tetraploid complementation). Two-cell embryos after nuclear transfer were transferred to the oviducts of surrogate mothers. Four (1.5%) of 272 nuclear-transferred two-cell embryos developed to term, and two (0.7%) developed into fertile adults. Nineteen (1.9%) of 992 tetraploid blastocysts receiving ES cells reached term, and 10 (1.0%) developed into adults. These findings demonstrate that some of the nuclei of differentiated neurons in the cerebral cortex of juvenile mice maintain developmental pluripotency.

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Adult murine neurons: their chromatin and chromosome changes and failure to support embryonic development as revealed by nuclear transfer

Cited by 40 publications

References 38 publications

Aneuploid neurons are functionally active and integrated into brain circuitry

Aneuploid neurons are functionally active and integrated into brain circuitry

Donor cell differentiation, reprogramming, and cloning efficiency: Elusive or illusive correlation?

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

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