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First published online February 24, 2006
doi: 10.1242/10.1242/dev.02283

Development 133, 1113-1123 (2006)
Published by The Company of Biologists 2006
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Direct visualization of nucleogenesis by precerebellar neurons: involvement of ventricle-directed, radial fibre-associated migration

Daisuke Kawauchi1,2,*, Hiroki Taniguchi3,{dagger}, Haruyasu Watanabe1, Tetsuichiro Saito4,* and Fujio Murakami1,{ddagger}

1 Laboratory of Neuroscience, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan.
2 SORST, Japan Science and Technology, Kawaguchi, Saitama 332-0012, Japan.
3 Division of Behavior and Neurobiology, National Institute for Basic Biology, Myodaiji-cho, Okazaki 444-8585, Japan.
4 Department of Development and Differentiation, Institute for Frontier Medical Sciences, Kyoto University, Kyoto 606-8507, Japan.

{ddagger} Author for correspondence (e-mail: murakami{at}fbs.osaka-u.ac.jp)

Accepted 13 January 2006

Nuclei are aggregates of neurons distributed in the central nervous system and are fundamental functional units that share anatomical and physiological features. Despite their importance, the cellular basis that leads to nucleogenesis is only poorly understood. Using exo utero electroporation with an enhanced yellow fluorescent protein (EYFP) gene, we show that the precerebellar neurons derived from the lower rhombic lip (lRL) undergo multiple migration steps to form nuclei. After the unilateral transfer of EYFP to the lRL of embryonic day 12.5 mice, EYFP-labelled neurons migrate tangentially from the lRL in two distinct streams, one towards the ventral metencephalon and the other towards the ventral myelencephalon. These neurons cross the ventral midline and then become radially directed. Labelled neurons in the tangential migratory streams form contralateral clusters in the external cuneate nucleus (ECN) and lateral reticular nucleus (LRN) in the myelencephalon, and bilateral clusters in the pontine grey nucleus (PGN) and reticulotegmental nucleus (RTN) in the metencephalon. Before forming the clusters, EYFP-labelled neurons begin to migrate radially towards the ventricle in close apposition to nestin-positive radial fibres, and then they aggregate as they detach from the fibres. Inhibition of cadherin function in ECN and LRN progenitors caused ipsilateral formation of the ECN and LRN, implying that the transition of their migration from tangential to radial involves a cell-intrinsic mechanism. These observations suggest that nucleogenesis of precerebellar neurons is a result of multi-phasic migration, and that ventricle-directed radial glia-guided migration is a key step for nucleogenesis.

Key words: Neuronal migration, Precerebellar neurons, Nucleogenesis, In vivo electroporation, Midline crossing, Ventricle-directed migration, Mouse


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