Dopamine D1 receptor interaction with arrestin3 in neostriatal neurons.
Dopamine D1 receptor interactions with arrestins have been characterized using heterologously expressed D1 receptor and arrestins. The purpose of this study was to investigate the interaction of the endogenous D1 receptor with endogenous arrestin2 and 3 in neostriatal neurons. Endogenous arrestin2 and 3 in striatal homogenates bound to the C-terminus of the D1 receptor in a glutathione-S-transferase (GST) pulldown assay, with arrestin3 binding more strongly. The D1 C-terminus and, to a lesser extent, the third cytoplasmic loop also bound purified arrestin2 and 3. In neostriatal neurons, 2, 5, and 20 min agonist treatment increased the colocalization of the D1 receptor and arrestin3 immunoreactivity without altering the colocalization of the D1 receptor and arrestin2. Further, agonist treatment for 5 and 20 min caused translocation of arrestin3, but not arrestin2, to the membrane. The binding of arrestin3, but not arrestin2, to the D1 receptor was increased as assessed by coimmunoprecipitation after agonist treatment for 5 and 20 min. Agonist treatment of neurons induced D1 receptor internalization (35-45%) that was maximal within 2-5 min, a time-course similar to that of the increase in colocalization of the D1 receptor with arrestin3. These data indicate that the D1 receptor preferentially interacts with arrestin3 in neostriatal neurons.
Macey
Tara A
TA
Department of Behavioral Neuroscience, Oregon Health & Science University and Veterans Affairs Medical Center, Portland, Oregon 97239-2999, USA.
Liu
Yong
Y
Gurevich
Vsevolod V
VV
Neve
Kim A
KA
D000818:Animals; D000831:Animals, Newborn; D019390:Arrestins / Q000378:metabolism*; D001552:Benzazepines / Q000494:pharmacology; D015153:Blotting, Western / Q000379:methods; D002478:Cells, Cultured; D018491:Dopamine Agonists / Q000494:pharmacology; D047468:Immunoprecipitation / Q000379:methods; D017072:Neostriatum / Q000166:cytology* / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism; D021381:Protein Transport / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D017447:Receptors, Dopamine D1 / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013997:Time Factors
D019390:Arrestins; D001552:Benzazepines; D018491:Dopamine Agonists; D010750:Phosphoproteins; D017447:Receptors, Dopamine D1; D011994:Recombinant Proteins; C082333:arrestin3; C071262:SK&F 82958