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Epithelial and endothelial expression of the green fluorescent protein reporter gene under the control of bovine prion protein (PrP) gene regulatory sequences in transgenic mice
- Lemaire-Vielle, C., Schulze, T., Podevin-Dimster, V., Follet, J., Bailly, Y., Blanquet-Grossard, F., Decavel, J.P., Heinen, E., Cesbron, J.Y.
- Proceedings of the National Academy of Sciences of the United States of America 2000 v.97 no.10 pp. 5422-5427
- recombinant DNA, animal proteins, Scyphozoa, promoter regions, small intestine, fluorescence, lymphocytes, gene expression, messenger RNA, alternative splicing, peripheral nerves, neurons, cattle, mice, kidneys, keratinocytes, exons, endothelium, medulla oblongata, thymus gland, transgenic animals, reporter genes, cerebellum
- The expression of the cellular form of the prion protein (PrPc) gene is required for prion replication and neuroinvasion in transmissible spongiform encephalopathies. The identification of the cell types expressing PrPc is necessary to understanding how the agent replicates and spreads from peripheral sites to the central nervous system. To determine the nature of the cell types expressing PrPc, a green fluorescent protein reporter gene was expressed in transgenic mice under the control of 6.9 kb of the bovine PrP gene regulatory sequences. It was shown that the bovine PrP gene is expressed as two populations of mRNA differing by alternative splicing of one 115-bp 5' untranslated exon in 17 different bovine tissues. The analysis of transgenic mice showed reporter gene expression in some cells that have been identified as expressing PrP, such as cerebellar Purkinje cells, lymphocytes, and keratinocytes. In addition, expression of green fluorescent protein was observed in the plexus of the enteric nervous system and in a restricted subset of cells not yet clearly identified as expressing PrP: the epithelial cells of the thymic medullary and the endothelial cells of both the mucosal capillaries of the intestine and the renal capillaries. These data provide valuable information on the distribution of PrPc at the cellular level and argue for roles of the epithelial and endothelial cells in the spread of infection from the periphery to the brain. Moreover, the transgenic mice described in this paper provide a model that will allow for the study of the transcriptional activity of the PrP gene promoter in response to scrapie infection.