Establishment of stably transfected rat neuronal cell lines expressing alpha-synuclein GFP fusion proteins.
Mutations in the alpha-synuclein gene have been linked to rare cases of familial Parkinson's disease (PD). alpha-Synuclein, a 140 amino acid polypeptide, is a major component of Lewy bodies (LB), a pathological hallmark of PD. Transgenic mice, Drosophila and marmosets (Challitrix jacchus) expressing either wild type (WT) or mutant human alpha-synuclein develop motor deficits, LB-like inclusions in some neurons and neuronal degeneration. The effects of human alpha-synuclein were investigated in a neuronal rat cell line (B103). Plasmids expressing WT and mutant human alpha-synuclein regulated by the cytomegalovirus (CMV) promoter were prepared and used for creating stably transfected neuronal rat cell lines. For localizing alpha-synuclein expression, stably transfected neuronal rat cell lines, expressing alpha-synuclein enhanced green fluorescent protein fusion proteins, regulated by either the CMV or the human platelet-derived growth factor ss promoter were generated. Over-expression of WT and A53T alpha-synuclein regulated by CMV promoter in stable transfectants resulted in formation of alpha-synuclein-immunopositive inclusion-like structures and mitochondrial alterations. Taken together, these results suggest that abnormal accumulation of alpha-synuclein could lead to mitochondrial alterations that might result in oxidative stress and eventually, cell death.
Schwach
Gert
G
JSW Life Sciences, Grambach, Austria.
Tschemmernegg
Michaela
M
Pfragner
Roswitha
R
Ingolic
Elisabeth
E
Schreiner
Edith
E
Windisch
Manfred
M
D000368:Aged; D000818:Animals; D002460:Cell Line / Q000502:physiology / Q000648:ultrastructure; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D051379:Mice; D009474:Neurons / Q000502:physiology / Q000648:ultrastructure; D010300:Parkinson Disease / Q000473:pathology / Q000503:physiopathology; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D051844:alpha-Synuclein / Q000235:genetics / Q000378:metabolism*
D011993:Recombinant Fusion Proteins; D051844:alpha-Synuclein; D049452:Green Fluorescent Proteins