Generation of Cloned Mice from Adult Neurons by Direct Nuclear Transfer1
| Autor: | Narumi Ogonuki, Mami Oikawa, Ryutaro Hirasawa, Hiroaki Nagatomo, Shogo Matoba, Kuniya Abe, Kimiko Inoue, Eiji Mizutani, Atsuo Ogura, Atsu Aiba, Satoshi Kamimura, Hirosuke Shiura, Hidetoshi Kassai, Teruhiko Wakayama |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
Cloning
Cerebellum medicine.drug_class Dentate gyrus Histone deacetylase inhibitor Cerebellar Purkinje cell Cell Biology General Medicine Biology Molecular biology Histone H3 Trichostatin A medicine.anatomical_structure nervous system Reproductive Medicine medicine Somatic cell nuclear transfer medicine.drug |
| Zdroj: | Biology of Reproduction. 92 |
| ISSN: | 1529-7268 0006-3363 |
| DOI: | 10.1095/biolreprod.114.123455 |
| Popis: | Whereas cloning mammals by direct somatic cell nuclear transfer has been successful using a wide range of donor cell types, neurons from adult brain remain ‘‘unclonable’’ for unknown reasons. Here, using a combination of two epigenetic approaches, we examined whether neurons from adult mice could be cloned. First, we used a specific antibody to discover cell types with reduced amounts of a repressive histone mark— dimethylated histone H3 lysine 9 (H3K9me2)—and identified CA1 pyramidal cells in the hippocampus and Purkinje cells in the cerebellum as candidates. Second, reconstructed embryos were treated with trichostatin A (TSA), a potent histone deacetylase inhibitor. Using CA1 cells, cloned offspring were obtained at high rates, reaching 10.2% and 4.6% (of embryos transferred) for male and female donors, respectively. Cerebellar Purkinje cell nuclei were too large to maintain their genetic integrity during nuclear transfer, leading to developmental arrest of embryos. However, gene expression analysis using cloned blastocysts corroborated a high rate of genomic reprogrammability of CA1 pyramidal and Purkinje cells. Neurons from the hippocampal dentate gyrus and cerebral cortex, which had higher amounts of H3K9me2, could also be used for producing cloned offspring, but the efficiencies were low. A more thorough analysis revealed that TSA treatment was essential for cloning adult neuronal cells. This study demonstrates, to our knowledge for the first time, that adult neurons can be cloned by nuclear transfer. Furthermore, our data imply that reduced amounts of H3K9me2 and increased histone acetylation appear to act synergistically to improve the development of cloned embryos assisted reproductive technology, CA1 pyramidal cells, cloning, developmental biology, early development, hippocampus, mouse, neural cells, nuclear transfer, reprogramming |
| Databáze: | OpenAIRE |
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