Characterization of cocaine and antidepressant-sensitive norepinephrine transporters in rat placental trophoblasts.
1. This paper reports on a primary cell culture system that predominantly expresses native norepinephrine (NE) transporters (NETs), and is amenable to biophysical as well as biochemical analyses. 2. Previous research has identified human and rat placentas as rich sources of NET. We have exploited this to develop primary cultures of rat placental trophoblasts. NE uptake in these cultures is about 10 times higher when compared to 5HT uptake. The presence of NET protein is revealed by immunoblot analysis, while there is no detectable SERT protein. 3. NE transport in rat trophoblasts is sensitive to NET-specific antagonists, desipramine (DS) and nisoxetine (NX), but not to the dopamine-transporter (DAT) specific antagonist, GBR12909 or to the serotonin (5HT) transporter (SERT) specific antagonist paroxetine (PX). Drugs of abuse such as cocaine and amphetamine also inhibit NE transport in these cells. Together these results suggest that rat placental trophoblasts predominantly express NET over other monoamine transporters. 4. Patch-clamp analysis reveals that NETs in intact rat trophoblasts are electrogenic. Comparison of NE uptake with NE-induced currents suggests that these two modes of transporter activity are differentially regulated.
Jayanthi
Lankupalle D
LD
Department of Pharmacology, Vanderbilt University School of Medicine, Center for Molecular Neuroscience, Nashville, Tennessee, TN 37232, USA. jayanthi@musc.edu
Vargas
Gricelly
G
DeFelice
Louis J
LJ
D000818:Animals; D000928:Antidepressive Agents / Q000378:metabolism*; D001665:Binding Sites; D002478:Cells, Cultured; D003042:Cocaine / Q000378:metabolism*; D018765:Dopamine Uptake Inhibitors / Q000378:metabolism; D005260:Female; D005473:Fluoxetine / Q000031:analogs & derivatives* / Q000378:metabolism; D008564:Membrane Potentials / Q000502:physiology; D009638:Norepinephrine / Q000037:antagonists & inhibitors / Q000378:metabolism* / Q000502:physiology; D050484:Norepinephrine Plasma Membrane Transport Proteins; D018408:Patch-Clamp Techniques; D011247:Pregnancy; D051381:Rats; D017207:Rats, Sprague-Dawley; D027981:Symporters / Q000096:biosynthesis / Q000378:metabolism*; D014327:Trophoblasts / Q000166:cytology / Q000378:metabolism*
D000928:Antidepressive Agents; D018765:Dopamine Uptake Inhibitors; D050484:Norepinephrine Plasma Membrane Transport Proteins; C493226:Slc6a2 protein, rat; D027981:Symporters; D005473:Fluoxetine; C011386:nisoxetine; D003042:Cocaine; D009638:Norepinephrine