Determination of Cellular Localization of Expression of Flavin-Containing Monooxygenase Genes in Mouse Tissues by In Situ Hybridization
Methods are described for the cellular localization of expression of flavin-containing monooxygenase (FMO) genes in various mouse tissues by in situ hybridization. These include the production of digoxigenin (DIG)-labeled antisense and sense RNA probes by transcription from FMO cDNA templates, the preparation of paraffin wax-embedded and cryostat tissue sections, the hybridization of RNA probes to tissue sections, and the specific detection of hybridized probes using an antibody to DIG.
Key WordsFlavin-containing monooxygenase antisense RNA probes in situ hybridization mouse digoxigenin-labeled probes gene expression
This work was supported by a grant from the Wellcome Trust (no. 053590).
- 5.Dolphin, C. T., Cullingford, T. E., Shephard, E. A., Smith, R. L., and Phillips, I. R. (1996) Differential developmental and tissue-specific regulation of expression of the genes encoding three members of the flavin-containing monooxygenase family of man, FMO1, FMO3 and FMO4. Eur. J. Biochem. 235, 683–689.CrossRefPubMedGoogle Scholar
- 16.Rodriguez, R. J. and Acosta, D. Jr. (1997) Metabolism of ketoconazole and deacetylated ketoconazole by rat hepatic microsomes and flavin-containing monooxygenases. Drug Metab, Dispos, 25, 772–777.Google Scholar
- 18.Wilkinson, D. G. (1992) The Theory and Practice of In Situ Hybridization, IRL, Oxford, UK, pp. 1–13.Google Scholar