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The Journal of Neuroscience, September 14, 2005, 25(37):8368-8374; doi:10.1523/JNEUROSCI.1591-05.2005

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Development/Plasticity/Repair
Developmental Pluripotency of the Nuclei of Neurons in the Cerebral Cortex of Juvenile Mice

Tomoharu Osada,^1,2 Nobuaki Tamamaki,³ Si-Young Song,⁴ Naoki Kakazu,⁵ Yukiko Yamazaki,⁶ Hatsune Makino,^1,2 Ayako Sasaki,^1,2 Teruyoshi Hirayama,^1,2 Shun Hamada,^1,2 Klaus-Armin Nave,⁷ Ryuzo Yanagimachi,⁶ and Takeshi Yagi^1,2,8

¹Core Research for Evolutional Science and Technology Research Agency, Japan Science and Technology Agency, Kawaguchi, Saitama 332-0012, Japan, and ²KOKORO Biology Group, Department of Integrated Biology, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan, ³Department of Morphological Neural Science, Graduate School of Medical Sciences, Kumamoto University, Kumamoto, Kumamoto 860-8556, Japan, ⁴Mitsubishi Kagaku Institute of Life Sciences, Machida, Tokyo 194-8511, Japan, ⁵Department of Molecular-Targeting Cancer Prevention, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, Kyoto 602-8566, Japan, ⁶The Institute for Biogenesis Research, Department of Anatomy and Reproductive Biology, John A. Burns School of Medicine, University of Hawaii, Honolulu, Hawaii 96822, ⁷Max Planck Institute of Experimental Medicine, Department of Neurogenetics, D-69120 Heidelberg, Germany, and ⁸Laboratory of Neurobiology and Behavioral Genetics, National Institute for Physiological Sciences, Okazaki, Aichi 444-8585, Japan

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.

Key words: pyramidal neurons; GABAergic neurons; cloning; nuclear transfer; embryonic stem (ES) cells; developmental plasticity

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Received April 22, 2005; revised July 29, 2005; accepted July 31, 2005.

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