Immunochemical analysis of dopamine transporter protein in Parkinson's disease.
The plasma membrane dopamine transporter (DAT) is considered to be a reliable marker of presynaptic dopaminergic terminal loss. Previous in vivo imaging and postmortem binding studies have detected a loss in striatal DAT binding in Parkinson's diseased (PD) brain; however, these techniques have poor spatial resolution and may suffer from nonspecific binding of some ligands. In this study, we use novel highly specific monoclonal antibodies to distinct epitopes of human DAT to quantify and localize the protein. Western blot analysis revealed marked reductions in DAT immunoreactivity in putamen, caudate, and nucleus accumbens of PD brain compared with control cases, and the reductions were significantly correlated to disease duration. Immunohistochemistry revealed DAT-immunoreactive fibers and puncta that were dense throughout the striatum of control brains but that were drastically reduced in putamen of PD brains. Caudate from PD brains showed a significant degree of sparing along the border of the ventricle, and the nucleus accumbens was relatively preserved. An unexpected finding was that discrete islands of DAT immunoreactivity were preserved within the matrix of PD putamen. Thus, immunological analysis of DAT protein provides novel and sensitive means for localizing and quantifying DAT protein in PD and other neurological disorders involving dopaminergic systems.
Miller
G W
GW
Department of Neurology, Emory University School of Medicine, Atlanta, GA 30322, USA.
Staley
J K
JK
Heilman
C J
CJ
Perez
J T
JT
Mash
D C
DC
Rye
D B
DB
Levey
A I
AI
D000368:Aged; D000369:Aged, 80 and over; D000911:Antibodies, Monoclonal / Q000032:analysis; D002352:Carrier Proteins / Q000032:analysis*; D002421:Caudate Nucleus / Q000737:chemistry; D003342:Corpus Striatum / Q000737:chemistry; D050483:Dopamine Plasma Membrane Transport Proteins; D005260:Female; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry / Q000379:methods; D008297:Male; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D008875:Middle Aged; D009419:Nerve Tissue Proteins / Q000032:analysis*; D010300:Parkinson Disease / Q000473:pathology*; D011699:Putamen / Q000737:chemistry; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D050483:Dopamine Plasma Membrane Transport Proteins; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D009419:Nerve Tissue Proteins; C493214:SLC6A3 protein, human