Selective transport of monoamine neurotransmitters by human plasma membrane monoamine transporter and organic cation transporter 3.
The plasma membrane monoamine transporter (PMAT) and organic cation transporter 3 (OCT3) are the two most prominent low-affinity, high-capacity (i.e., uptake(2)) transporters for endogenous biogenic amines. Using the Flp-in system, we expressed human PMAT (hPMAT) and human OCT3 (hOCT3) at similar levels in human embryonic kidney 293 cells. Parallel and detailed kinetics analysis revealed distinct and seemingly complementary patterns for the two transporters in transporting monoamine neurotransmitters. hPMAT is highly selective toward serotonin (5-HT) and dopamine, with the rank order of transport efficiency (V(max)/K(m)) being: dopamine, 5-HT ≫ histamine, norepinephrine, epinephrine. The substrate preference of hPMAT toward these amines is substantially driven by large (up to 15-fold) distinctions in its apparent binding affinities (K(m)). In contrast, hOCT3 is less selective than hPMAT toward the monoamines, and the V(max)/K(m) rank order for hOCT3 is: histamine > norepinephrine, epinephrine > dopamine >5-HT. It is noteworthy that hOCT3 demonstrated comparable (≤2-fold difference) K(m) toward all amines, and distinctions in V(max) played an important role in determining its differential transport efficiency toward the monoamines. Real-time reverse transcription-polymerase chain reaction revealed that hPMAT is expressed at much higher levels than hOCT3 in most human brain areas, whereas hOCT3 is selectively and highly expressed in adrenal gland and skeletal muscle. Our results suggest that hOCT3 represents a major uptake(2) transporter for histamine, epinephrine, and norepinephrine. hPMAT, on the other hand, is a major uptake(2) transporter for 5-HT and dopamine and may play a more important role in transporting these two neurotransmitters in the central nervous system.
Duan
Haichuan
H
Department of Pharmaceutics, University of Washington, H272J Health Science Bldg, Seattle, WA 98195, USA.
Wang
Joanne
J
D015655:1-Methyl-4-phenylpyridinium / Q000378:metabolism; D000311:Adrenal Glands / Q000378:metabolism; D000818:Animals; D015306:Biogenic Monoamines / Q000378:metabolism*; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology*; D001921:Brain / Q000378:metabolism; D004254:DNA Nucleotidyltransferases / Q000235:genetics; D004298:Dopamine / Q000378:metabolism; D004837:Epinephrine / Q000378:metabolism; D033705:Equilibrative Nucleoside Transport Proteins / Q000235:genetics / Q000378:metabolism*; D005260:Female; D015870:Gene Expression / Q000235:genetics; D057809:HEK293 Cells; D006632:Histamine / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008297:Male; D026901:Membrane Transport Proteins / Q000378:metabolism; D051379:Mice; D008815:Mice, Inbred Strains; D018482:Muscle, Skeletal / Q000378:metabolism; D018377:Neurotransmitter Agents / Q000378:metabolism*; D009638:Norepinephrine / Q000378:metabolism; D027701:Organic Cation Transport Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D012701:Serotonin / Q000378:metabolism; D013574:Synaptosomes / Q000187:drug effects / Q000378:metabolism; D019789:Tetraethylammonium / Q000378:metabolism; D014162:Transfection; D000099254:Organic Cation Transporter 3
D015655:1-Methyl-4-phenylpyridinium; D015306:Biogenic Monoamines; D004254:DNA Nucleotidyltransferases; D004298:Dopamine; D004837:Epinephrine; D033705:Equilibrative Nucleoside Transport Proteins; D006632:Histamine; D026901:Membrane Transport Proteins; D018377:Neurotransmitter Agents; D009638:Norepinephrine; D027701:Organic Cation Transport Proteins; D012701:Serotonin; D019789:Tetraethylammonium; D000099254:Organic Cation Transporter 3; C495988:SLC29A4 protein, human; C513828:Slc29a4 protein, mouse; C045119:FLP recombinase