RESTORATION OF HOLOCERULOPLASMIN SYNTHESIS IN LEC RAT AFTER INFUSION OF RECOMBINANT ADENOVIRUS BEARING WND CDNA

Wilson's disease, an autosomal recessive disorder, is characterized by the excessive accumulation of copper in the liver. WND (ATP7B) gene, which encodes a putative copper transporting P-type ATPase, is defecti ve in the patients, To investigate the in vivo function of WND protein as well as its intracellular localization, WND cDNA was introduced to the Long-Evans Cinnamon rat, known as a rodent model for Wilson's dis ease, by recombinant adenovirus-mediated gene delivery. An immunofluor escent study and a subcellular fractionation study revealed the transg ene expression in liver and its localization to the Golgi apparatus, M oreover, since the synthesis of holoceruloplasmin is disturbed in the Long-Evans Cinnamon rat, the plasma level of holoceruloplasmin, oxidas e-active and copper-bound form, was examined to evaluate the function of WND protein with respect to the copper transport. Consequently, the appearance of holoceruloplasmin in plasma was confirmed by Western bl ot analysis and plasma measurements for the oxidase activity and the c opper content, These findings indicate that introduced WND protein may function in the copper transport coupled with the synthesis of cerulo plasmin and that the Gels apparatus is the likely site for WND protein to manifest its function.