Skip to main content
. 2014 Oct 14;4(10):e464. doi: 10.1038/tp.2014.90

Figure 3.

Figure 3

Intracellular AMPH delivery induces trafficking in hDAT A559V cells measured by transient charge movement. (a) Cells were voltage clamped in the whole-cell configuration at a baseline of −20 mV and the current recorded following a voltage step to −140 mV. DAT-mediated currents were defined by subtracting the nonspecific current obtained in the presence of the DAT blocker cocaine (10 μM) from those recorded in the absence of cocaine. Top, DAT-mediated control currents (dotted lines) before AMPH treatment are plotted and compared with currents recorded after AMPH treatment (10 μM AMPH for 10 min, solid lines). Extracellular AMPH induces DAT trafficking (compare AMPH solid lines with control dotted lines) in hDAT but not hDAT A559V cells. Bottom, the transient charge (Q) was obtained by integrating the area under transient current as an index of the number of transporters at the cell surface. AMPH significantly decreases Q in hDAT cells (*P<0.05, QAMPH vs Qcontrol Student's t-test, n=5) but not hDAT A559V cells (P>0.05, QAMPH vs Qcontrol Student's t-test, n=3). (b) AMPH was applied intracellularly through perfusion by the whole-cell patch clamp pipette internal solution. Top, DAT-mediated control currents immediately after achieving whole-cell access (dotted lines) are plotted compared with currents after perfusing the cell for 10 min with AMPH (10 μM, solid lines). AMPH does induce DAT trafficking of hDAT A559V cells comparable to that of hDAT cells. Bottom, AMPH significantly decreases Q in both hDAT and hDAT A559V cells (*P<0.05, QAMPH vs Qcontrol Student's t-test, n=3). AMPH, amphetamine; DAT, dopamine transporter; hDAT, human dopamine transporter.