Expression of the extraneuronal monoamine transporter in RPE and neural retina.
OBJECTIVE
Dopamine has several important functions in the retina including a possible role in controlling photoreceptor disk shedding to the RPE. While some cells express a transporter for dopamine, the RPE cell does not, leading us to ask whether the newly described catecholamine transport system, the extraneuronal monoamine transporter (uptake(2)) (also known as organic cation transporter 3 (OCT3), is present in RPE and might function as a transporter for dopamine. OCT3 also accepts histamine as a transportable substrate and so we investigated the interaction of this retinal neurotransmitter with OCT3.


METHODS
OCT3 expression in the mouse eye was analyzed by in situ hybridization, Northern blot analysis and RT-PCR. OCT3 function was analyzed in cultured human ARPE-19 cells by monitoring the uptake of 1-methyl-4-phenyl pyridinium (MPP(+)), a neurotoxin, which is a known substrate for OCT3.


RESULTS
In situ hybridization analysis showed that OCT3 is expressed in mouse RPE and in several cell types of the neural retina, including photoreceptor, ganglion, amacrine, and horizontal cells. The expression of OCT3 in RPE was confirmed by Northern blot analysis and RT-PCR. The characteristics of MPP( +) uptake in cultured ARPE-19 cells included the stimulation of transport by alkaline pH, high affinity (K(t) = 28 +/- 4 microM), competition with several cationic drugs and monoamine neurotransmitters and sensitivity to steroids. In addition, the uptake of MPP(+) in RPE cells was inhibited by dopamine and histamine with IC(50) values (concentration needed for 50% inhibition) of 637 +/- 84 microM and 150 +/- 20 microM, respectively. CONCLUSIONS. This study provides the first report on the expression and function of an organic cation transporter, OCT3, in the eye and in particular the RPE. The data have physiological and pharmacological relevance as it is likely that OCT3 participates in the clearance of dopamine and histamine from the subretinal space and may also play a key role in the disposition of the retinal neurotoxin MPP(+).
Rajan
P D
PD
Department of Biochemistry and Molecular Biology, Medical College of Georgia, Augusta, USA.
Kekuda
R
R
Chancy
C D
CD
Huang
W
W
Ganapathy
V
V
Smith
S B
SB
D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine / Q000494:pharmacology; D015655:1-Methyl-4-phenylpyridinium / Q000493:pharmacokinetics; D000661:Amphetamine / Q000494:pharmacology; D000818:Animals; D015306:Biogenic Monoamines / Q000494:pharmacology; D015152:Blotting, Northern; D002352:Carrier Proteins / Q000235:genetics*; D002478:Cells, Cultured; D005123:Eye / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D006801:Humans; D006863:Hydrogen-Ion Concentration; D017403:In Situ Hybridization; D007700:Kinetics; D008694:Methamphetamine / Q000494:pharmacology; D051379:Mice; D008813:Mice, Inbred ICR; D009538:Nicotine / Q000494:pharmacology; D027701:Organic Cation Transport Proteins; D010857:Pigment Epithelium of Eye / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D011342:Procainamide / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012160:Retina / Q000378:metabolism*; D013256:Steroids / Q000494:pharmacology; D019789:Tetraethylammonium / Q000494:pharmacology; D013997:Time Factors; D000099254:Organic Cation Transporter 3
D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; D015655:1-Methyl-4-phenylpyridinium; D000661:Amphetamine; D015306:Biogenic Monoamines; D002352:Carrier Proteins; D008694:Methamphetamine; D009538:Nicotine; D027701:Organic Cation Transport Proteins; D011342:Procainamide; D012333:RNA, Messenger; D013256:Steroids; D019789:Tetraethylammonium; D000099254:Organic Cation Transporter 3