Aim: 

Several reports established a relationship between systemic copper imbalance and the onset of invalidating pathologies of the central nervous system. To contribute to comprehension of such a link, we experimented a situation of copper impairment in a rat neuroblastoma cell model, B104, by using trientine, a well-known copper chelating agent clinically used for treatment of Wilson's disease.

Methods: 

Cells were routinely cultured in a medium supplemented with trientine for 48 and 96 hours. Transcript and protein levels of potentially copper-conditioned genes (CTR1, DCT1, ATP7A, PrP^C) were assessed in response to cell ion impairment by Real Time PCR and Western Blotting analysis. A fluorimetric method has been employed for a comparative estimation of the rate of copper influx in trientine-treated cells.

Results: 

There was no effect of trientine on CTR1, DCT1 and ATP7A mRNA and protein levels, while PrP^C gene expression was elevated after both 48 (p < 0.05) and 96 (p < 0.01) hour exposure of B104 cells to copper deprivation. Catalytic efficiency (V_max/K_m) of cell copper transport for the shorter treatment was greater than calculated for the control condition. For the longer period of treatment a dramatic reduction of both the maximal velocity and the K_m constant was observed, leaving the ratio substantially unaltered, presumably as a consequence of a moderate cell loss.

Conclusion: 

Experimental data demonstrate that in B104 neuroblastoma cell line copper status modulates the expression of prion protein gene in a time-dependent way, with presumed effects on kinetics of cell ion import.