Efficient adenoviral vector-directed expression of a foreign gene to neurons and sustentacular cells in the mouse olfactory neuroepithelium
| Autor: | Anthony Holtmaat, Michael G. Kaplitt, A. Beate Oestreicher, Willem Hendrik Gispen, Wim T. J. M. C. Hermens, Joost Verhaagen |
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| Rok vydání: | 1996 |
| Předmět: |
Male
Olfactory system Recombinant Fusion Proteins Transgene Genetic Vectors Cytomegalovirus Transfection Olfactory Receptor Neurons Viral vector Mice Cellular and Molecular Neuroscience Olfactory Mucosa Genes Reporter medicine Animals Infusions Parenteral Promoter Regions Genetic Molecular Biology Administration Intranasal Reporter gene biology Adenoviruses Human Stem Cells Genetic transfer Defective Viruses beta-Galactosidase Virology Cell biology medicine.anatomical_structure biology.protein Expression cassette Olfactory marker protein Olfactory epithelium |
| Zdroj: | Molecular Brain Research. 41:148-156 |
| ISSN: | 0169-328X |
| DOI: | 10.1016/0169-328x(96)00085-x |
| Popis: | Replication deficient recombinant adenoviral vectors are efficient gene transfer agents for postmitotic cells, including neurons and glial cells. In this paper we have examined the effectiveness of adenoviral vector-mediated gene transfer to the olfactory epithelium of adult mice. We show that Ad-LacZ, a prototype first generation adenoviral vector containing an expression cassette for the reporter gene LacZ, directs transgene expression to mature and immature olfactory neurons and to sustentacular cells. The technique to apply the vector to the nasal cavity and the amount of viral vector per mouse are important variables that determine the success of viral vector-mediated gene transfer to the mouse olfactory neuroepithelium. Slow infusion of the viral vector solution in fully anaesthetized mice yields the best result in terms of the number of epithelial cells transduced. Infection of the olfactory neuroepithelium with a moderate amount of viral vector (10(9) plaque-forming units (PFU)) results in transgene expression in many cells throughout the epithelium for 8-12 days, followed by a decline in transduced cells at 25 days postinstillation of the virus This decrement in transgene expression is consistent with the natural turnover process that occurs in the epithelium throughout adulthood. At high viral loads (1.3 x 10(10) PFU) extinction of transgene expression occurs as early as 8 days postinjection and is accompanied by epithelial degeneration indicating that the vector dose used should be carefully chosen. Taken together, the current observations demonstrate that adenoviral vectors are effective tools to genetically modify the adult mouse olfactory neuroepithelium in vivo. |
| Databáze: | OpenAIRE |
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