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The Journal of Neuroscience, January 7, 2004, 24(1):286-296; doi:10.1523/JNEUROSCI.2529-03.2004

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Development/Plasticity/Repair
Neuronal Birthdate-Specific Gene Transfer with Adenoviral Vectors

Mitsuhiro Hashimoto1 and Katsuhiko Mikoshiba1,2

1Laboratory for Developmental Neurobiology, RIKEN Brain Science Institute, 2-1 Hirosawa, Wako-shi, Saitama 351-0198, Japan, and 2Department of Molecular Neurobiology, The Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan

The multilayered structure of the cerebral cortex has been studied in detail. Early-born neurons migrate into the inner layer and late-born neurons migrate into more superficial layers, thus establishing an inside-out gradient. The progenitor cells appear to acquire layer-specific properties at the time of neuronal birth; however, the molecular mechanisms of cell-fate acquisition are still unclear, because it has been difficult to identify a cohort of birthdate-related progenitor cells. Using replication-defective adenoviral vectors, we successfully performed "pulse gene transfer" into progenitor cells in a neuronal birthdate-specific manner. When adenoviral vectors were injected into the midbrain ventricle of mouse embryos between embryonic day 10.5 (E10.5) and E14.5, the adenoviral vectors introduced a foreign gene into a specific cohort of birthdate-related progenitor cells. The virally infected cohorts developed normally into cortical neurons and formed the canonical cortical layers in an inside-out manner. This technique allows us to distinguish a cohort of birthdate-related progenitor cells from other progenitor cells with different birthdates and to introduce a foreign gene into specific subsets of cortical layers by performing adenoviral injection at specific times. This adenovirus-meditated gene transfer technique will enable us to examine the properties of each subset of progenitor cells that share the same neuronal birthdate.

Key words: neuronal birthdate; gene transfer; cerebral cortex; adenoviral vector; development; laminar structure

Received May 21, 2003; revised November 6, 2003; accepted November 6, 2003.




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