TAILIEUCHUNG - Báo cáo khoa học: Cell type-specific transgene expression of the prion protein in Xenopus intermediate pituitary cells

The cellular form of prion protein (PrP C ) is anchored to the plasma mem-brane of the cell and expressed in most tissues, but predominantly in the brain, including in the pituitary gland. Thus far, the biosynthesis of PrP C has been studied only in cultured (transfected) tumour cell lines and not in primary cells. | ềFEBS Journal Cell type-specific transgene expression of the prion protein in Xenopus intermediate pituitary cells Jos W. G. van Rosmalen and Gerard J. M. Martens Department of Molecular AnimalPhysiology Nijmegen Center for Molecular Life Sciences and Institute for Neuroscience Radboud University Nijmegen the Netherlands Keywords intermediate pituitary melanotrope cell post-translationalmodification prion protein biosynthesis transgenesis Xenopus laevis Correspondence . Martens Department of Molecular AnimalPhysiology Nijmegen Center for Molecular Life Sciences NCMLS and Institute for Neuroscience Radboud University Nijmegen Geert Grooteplein Zuid 28 6525 GA Nijmegen the Netherlands Fax 31 24 3615317 Tel 31 24 3610564 E-mail Received 5 October 2005 revised 20 December 2005 accepted 22 December 2005 doi The cellular form of prion protein PrPC is anchored to the plasma membrane of the cell and expressed in most tissues but predominantly in the brain including in the pituitary gland. Thus far the biosynthesis of PrPC has been studied only in cultured transfected tumour cell lines and not in primary cells. Here we investigated the intracellular fate of PrPC in vivo by using the neuroendocrine intermediate pituitary melanotrope cells of the South-African claw-toed frog Xenopus laevis as a model system. These cells are involved in background adaptation of the animal and produce high levels of its major secretory cargo proopiomelanocortin POMC when the animal is black-adapted. The technique of stable Xenopus transgenesis in combination with the POMC gene promoter was used as a tool to express Xenopus PrPC amino-terminally tagged with the green fluorescent protein GFP-PrPC specifically in the melanotrope cells. The GFP-PrPC fusion protein was expressed from stage-25 tadpoles onwards to juvenile frogs the expression was induced on a black background and the fusion protein was subcellularly located mainly in the .

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