title: Interspecies comparison of the functional characteristics of plasma membrane monoamine transporter (PMAT) between human, rat and mouse.
abstract: Plasma membrane monoamine transporter (PMAT) is a newly discovered monoamine transporter belonging to the equilibrative nucleoside transporter family. Highly expressed in the brain, PMAT represents a major uptake2transporter that may play a role in monoamine clearance. Although human PMAT has been functionally characterized at the molecular level, rodent models are often used to evaluate PMAT function in ex vivo and in vivo studies. The aim of this study was to examine if there is potential species difference in the functional characteristics of PMAT between human, rat and mouse. A set of transfected cells stably expressing human PMAT (MDCK/hPMAT), rat Pmat (MDCK/rPmat) and mouse Pmat (Flp293/mPmat) were constructed. In MDCK/hPMAT, MDCK/rPmat and Flp293/mPmat cells, cellular localization analyses revealed that hPMAT, rPmat and mPmat are expressed and mainly localized to the plasma membranes of cells. The uptake of MPP+, serotonin and dopamine by MDCK/hPMAT, MDCK/rPmat and Flp293/mPmat cells was significantly increased compared with those by the mock transfection control. In contrast, two nucleosides, uridine and adenosine, minimally interacted with PMAT/Pmat in all species. The hPMAT-, rPmat- and mPmat-mediated uptakes of MPP+, serotonin and dopamine were saturable, with Kmvalues of 33.7μM, 70.2μM and 49.5μM (MPP+), 116μM, 82.9μM and 231μM (serotonin), and 201μM, 271μM and 466μM (dopamine), respectively, suggesting similar substrate affinities between human and rodent PMAT/Pmat. The prototypical inhibitors, decynium 22 and GBR12935, also showed similar inhibition potencies between species. In conclusion, the present study demonstrated interspecies similarities in the functional characteristics of human and rodent PMAT/Pmat, which indicate a practical utility of rat and mouse animal models for further investigating and extrapolating the in vivo function of PMAT in humans.
authors.affiliation: Department of Pharmaceutics, School of Pharmacy, University of Washington, H272 Health Sciences Building, Seattle, WA 98195-7610, USA; Department of Biopharmaceutics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo, 192-0392, Japan.
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Shirasaka
authors.affiliation: Department of Pharmaceutics, School of Pharmacy, University of Washington, H272 Health Sciences Building, Seattle, WA 98195-7610, USA.
authors.forename: Nora
authors.initials: N
authors.lastname: Lee
authors.affiliation: Department of Pharmaceutics, School of Pharmacy, University of Washington, H272 Health Sciences Building, Seattle, WA 98195-7610, USA.
authors.forename: Haichuan
authors.initials: H
authors.lastname: Duan
authors.affiliation: Department of Pharmaceutics, School of Pharmacy, University of Washington, H272 Health Sciences Building, Seattle, WA 98195-7610, USA.
authors.forename: Horace
authors.initials: H
authors.lastname: Ho
authors.affiliation: Department of Pharmaceutics, School of Pharmacy, University of Washington, H272 Health Sciences Building, Seattle, WA 98195-7610, USA.
authors.forename: Joanna
authors.initials: J
authors.lastname: Pak
authors.affiliation: Department of Pharmaceutics, School of Pharmacy, University of Washington, H272 Health Sciences Building, Seattle, WA 98195-7610, USA. Electronic address: jowang@u.washington.edu.
authors.forename: Joanne
authors.initials: J
authors.lastname: Wang
mesh_terms: D000818:Animals; D006801:Humans; D051379:Mice; D050480:Plasma Membrane Neurotransmitter Transport Proteins / Q000378:metabolism*; D051381:Rats; D013045:Species Specificity
chemical_list: D050480:Plasma Membrane Neurotransmitter Transport Proteins
keywords: Human; Monoamine; Mouse; PMAT; Rat; Species difference