TY - JOUR
T1 - Salvinorin A regulates dopamine transporter function via a kappa opioid receptor and ERK1/2-dependent mechanism
AU - Kivell, Bronwyn
AU - Uzelac, Zeljko
AU - Sundaramurthy, Santhanalakshmi
AU - Rajamanickam, Jeyaganesh
AU - Ewald, Amy
AU - Chefer, Vladimir
AU - Jaligam, Vanaja
AU - Bolan, Elizabeth
AU - Simonson, Bridget
AU - Annamalai, Balasubramaniam
AU - Mannangatti, Padmanabhan
AU - Prisinzano, Thomas E.
AU - Gomes, Ivone
AU - Devi, Lakshmi A.
AU - Jayanthi, Lankupalle D.
AU - Sitte, Harald H.
AU - Ramamoorthy, Sammanda
AU - Shippenberg, Toni S.
N1 - Funding Information:
This work was supported by the National Institutes of Health and National Institute on Drug Abuse Intramural Research Program (T.S.S), NIH grants, MH083928 , MH091633 (S.R), DA018151 (T.E.P), DA019521 , DA08863 , GM071558 (L.A.D.), The Health Research Council of New Zealand (B.K) and Austrian Science Fund/FWF , P23658 (H.H.S). CCRP-YFP was kindly provided by Dr Michel Bouvier (University of Montreal). DAT constructs were provided by Dr. Jonathan Javitch.
PY - 2014/8/19
Y1 - 2014/8/19
N2 - Salvinorin A (SalA), a selective κ-opioid receptor (KOR) agonist, produces dysphoria and pro-depressant like effects. These actions have been attributed to inhibition of striatal dopamine release. The dopamine transporter (DAT) regulates dopamine transmission via uptake of released neurotransmitter. KORs are apposed to DAT in dopamine nerve terminals suggesting an additional target by which SalA modulates dopamine transmission. SalA produced a concentration-dependent, nor-binaltorphimine (BNI)- and pertussis toxin-sensitive increase of ASP+ accumulation in EM4 cells coexpressing myc-KOR and YFP-DAT, using live cell imaging and the fluorescent monoamine transporter substrate, trans 4-(4-(dimethylamino)-styryl)-N- methylpyridinium) (ASP+). Other KOR agonists also increased DAT activity that was abolished by BNI pretreatment. While SalA increased DAT activity, SalA treatment decreased serotonin transporter (SERT) activity and had no effect on norepinephrine transporter (NET) activity. In striatum, SalA increased the Vmax for DAT mediated DA transport and DAT surface expression. SalA up-regulation of DAT function is mediated by KOR activation and the KOR-linked extracellular signal regulated kinase- (ERK1/2) pathway. Co-immunoprecipitation and BRET studies revealed that DAT and KOR exist in a complex. In live cells, DAT and KOR exhibited robust FRET signals under basal conditions. SalA exposure caused a rapid and significant increase of the FRET signal. This suggests that the formation of KOR and DAT complexes is promoted in response to KOR activation. Together, these data suggest that enhanced DA transport and decreased DA release resulting in decreased dopamine signalling may contribute to the dysphoric and pro-depressant like effects of SalA and other KOR agonists.
AB - Salvinorin A (SalA), a selective κ-opioid receptor (KOR) agonist, produces dysphoria and pro-depressant like effects. These actions have been attributed to inhibition of striatal dopamine release. The dopamine transporter (DAT) regulates dopamine transmission via uptake of released neurotransmitter. KORs are apposed to DAT in dopamine nerve terminals suggesting an additional target by which SalA modulates dopamine transmission. SalA produced a concentration-dependent, nor-binaltorphimine (BNI)- and pertussis toxin-sensitive increase of ASP+ accumulation in EM4 cells coexpressing myc-KOR and YFP-DAT, using live cell imaging and the fluorescent monoamine transporter substrate, trans 4-(4-(dimethylamino)-styryl)-N- methylpyridinium) (ASP+). Other KOR agonists also increased DAT activity that was abolished by BNI pretreatment. While SalA increased DAT activity, SalA treatment decreased serotonin transporter (SERT) activity and had no effect on norepinephrine transporter (NET) activity. In striatum, SalA increased the Vmax for DAT mediated DA transport and DAT surface expression. SalA up-regulation of DAT function is mediated by KOR activation and the KOR-linked extracellular signal regulated kinase- (ERK1/2) pathway. Co-immunoprecipitation and BRET studies revealed that DAT and KOR exist in a complex. In live cells, DAT and KOR exhibited robust FRET signals under basal conditions. SalA exposure caused a rapid and significant increase of the FRET signal. This suggests that the formation of KOR and DAT complexes is promoted in response to KOR activation. Together, these data suggest that enhanced DA transport and decreased DA release resulting in decreased dopamine signalling may contribute to the dysphoric and pro-depressant like effects of SalA and other KOR agonists.
KW - Dopamine transporter
KW - Dysphoric
KW - Kappa opioid receptor
KW - Pro-depressant
KW - Salvinorin A
KW - Serotonin transporter
KW - Trafficking
UR - http://www.scopus.com/inward/record.url?scp=84907359962&partnerID=8YFLogxK
U2 - 10.1016/j.neuropharm.2014.07.016
DO - 10.1016/j.neuropharm.2014.07.016
M3 - Article
C2 - 25107591
AN - SCOPUS:84907359962
SN - 0028-3908
VL - 86
SP - 228
EP - 240
JO - Neuropharmacology
JF - Neuropharmacology
ER -