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The effect of paraquat on the expression of D2 dopamine receptor in CAD cells
Author(s) -
Kim OkJin J,
Megaris Michael Paul,
Azadani Zahra Crystal
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.717.6
Subject(s) - hek 293 cells , western blot , microbiology and biotechnology , radioligand , dopamine receptor d2 , proximity ligation assay , transfection , chemistry , biology , receptor , cell culture , gene , biochemistry , genetics
We identified a protein kinase Czeta interacting protein (ZIP1) as an interacting protein of the D2 dopamine receptor (DAR) through a yeast two hybrid assay. We confirmed that the functional interaction between D2 DAR and ZIP1 in human embryonic kidney 293 (HEK 293) cells by using immunoprecipitation/western blot analysis, cAMP assays and radioligand binding assays. Significantly, we also demonstrated that D2 DAR and ZIP1 were coimmunoprecipitated from mouse brain and colocalized in mouse brain. The role of ZIP1 has not been clearly elucidated although overexpression of ZIP1 resulted in the reduction of membrane associated D2 DAR in HEK 293T cells. Our studies also demonstrated that ZIP1 was induced by an herbicide paraquat in HEK 293T cells and Cath.a.Differentiated (CAD) dopaminergic neuronal cells. The expression of membrane associated D2 DAR was reduced in CAD cells stably transfected with D2 DAR (CAD‐D2 DAR) in the presence of paraquat by radioligand binding assays. Subsequent radioligand binding assay and Western blot analysis studies also appeared to indicate that paraquat reduces the expression of membrane associated D2 DAR in the presence of ZIP1 in CAD‐D2 DAR cells. The further characterization of the effect of paraquat on the expression and trafficking of D2 DAR are still in the progress. This work was supported by the University of Kansas General Research Fund to OJK and Undergraduate Research Assistantship fund to MM.