title: Membrane potential and pH-dependent accumulation of decynium-22 (1,1'-diethyl-2,2'-cyanine iodide) flourencence through OCT transporters in astrocytes.
abstract: 1,1 '-Diethyl-2,2'-cyanine iodide (decynium22; D22) is a potent blocker of the organic cation family of transporters (EMT/OCT) known to move endogenous monoamines like dopamine and norepinephrine across cell membranes. Decynium22 is a cation with a relatively high affinity for all members of the OCT family in both human and rat cells. The mechanism through which decynium22 blocks OCT transporters are poorly understood. We tested the hypothesis that denynium22 may compete with monoamines utilizing OCT to permeate the cells. Using the ability of D22 to aggregate and produce fluorescence at 570 nm, we measured D22 uptake in cultured astrocytes. The rate of D22 uptake was strongly depressed by acid pH and by elevated external K+. The rate of uptake was similar to that displayed by 4-(4-(dimethylamino)-styryl)-N-methylpyridinium (ASP+), a well established substrate for OCT and high-affinity Na+-dependent monoamine transporters. These data were supported by measurement of electrogenic uptake using whole cell voltage clamp recording. Decynium22 depressed norepinephrine, but not glutamate uptake. These data are also consistent with the described OCT transporter characteristics. Taken together, our results suggest that decynium22 accumulation might be a useful instrument to study monoamine transport in the brain, and particularly in astrocytes, where they may play a prominent role in monoamine uptake during brain dysfunction related to monoamines (like Parkinson disease) and drug addiction.
authors.affiliation: Department of Physiology, Universidad Central del Caribe, School of Medicine, Bayamon, PR.
authors.forename: Mikhail
authors.initials: M
authors.lastname: Inyushin
authors.forename: Yuri
authors.initials: Y
authors.lastname: Kucheryaykh
authors.forename: Lilia
authors.initials: L
authors.lastname: Kucheryavykh
authors.forename: Priscilla
authors.initials: P
authors.lastname: Sanabria
authors.forename: Carlos
authors.initials: C
authors.lastname: Jiménez-Rivera
authors.forename: Irina
authors.initials: I
authors.lastname: Struganova
authors.forename: Misty
authors.initials: M
authors.lastname: Eaton
authors.forename: Serguei
authors.initials: S
authors.lastname: Skatchkov
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000502:physiology*; D002478:Cells, Cultured; D005453:Fluorescence; D006863:Hydrogen-Ion Concentration; D008564:Membrane Potentials; D027701:Organic Cation Transport Proteins / Q000502:physiology*; D011804:Quinolines / Q000378:metabolism*; D051381:Rats
chemical_list: D027701:Organic Cation Transport Proteins; D011804:Quinolines; C100254:pseudoisocyanine