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Acta Physiologica 2010; Volume 200, Supplement 681
Abstracts of the 61st National Congress of the Italian Physiological Society
9/15/2010-9/17/2010
Varese, Italy
REGULATORY MECHANISMS OF COPPER HOMEOSTASIS IN NEUROBLASTOMA CELLS: THE IMPORTANCE OF PRION PROTEIN
Abstract number: P17
URSO1 E, RIZZELLO1 A, DANIELI1 A, ACIERNO1 R, STORELLI1 C, MAFFIA1 M
1Dept Biological and Environmental Science and Technology, Salento Univ., Lecce, Italy
Neuronal functions critically depend on the maintenance of cell copper (Cu) homeostasis, but a full knowledge of the regulatory mechanisms has not been reached yet. Cu excess/deficiency is known to activate in vitro a transcriptional response, mainly involving the oxidative stress genes, but the effects on the kinetics of cell Cu import are largely unexplored. It's currently accepted that the integral membrane protein Ctr1 functions as the major Cu importer, but our previous studies demonstrated in a neuron-like cell model the involvement of the surface prion protein (PrPC) in driving cell Cu internalization. This work focused on the adaptive response of a rat neuroblastoma cell line (B104) subjected to an inadequate Cu supply, with special attention to PrPC role. Preliminarily, the health condition of Cu-depleted cells was investigated by metabolic assays (i.e. oxygen consumption rate), then the transcriptional response (Cu import proteins; CCS, Atox1 and Cox17 chaperons; ATP7A efflux pump) was analyzed by RT-PCR, and the more relevant results confirmed by proteomic studies. Remarkably, an increase in PrPC expression was detected in Cu-depleted cultures, associated to an enhanced cell Cu uptake activity, measured by a fluorimetric technique. This finding evidenced the importance of PrPC as a cell Cu importer under ion deficiency. Work is in progress to accurately quantify its contribution in Cu transport processes using PrPC knocked-down neuroblastoma cell models.
To cite this abstract, please use the following information:
Acta Physiologica 2010; Volume 200, Supplement 681 :P17