Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Evid Based Complement Alternat Med
2012 Jan 01;2012:818451. doi: 10.1155/2012/818451.
Show Gene links
Show Anatomy links
Inhibition of Activity of GABA Transporter GAT1 by δ-Opioid Receptor.
Pu L
,
Xu N
,
Xia P
,
Gu Q
,
Ren S
,
Fucke T
,
Pei G
,
Schwarz W
.
???displayArticle.abstract???
Analgesia is a well-documented effect of acupuncture. A critical role in pain sensation plays the nervous system, including the GABAergic system and opioid receptor (OR) activation. Here we investigated regulation of GABA transporter GAT1 by δOR in rats and in Xenopus oocytes. Synaptosomes of brain from rats chronically exposed to opiates exhibited reduced GABA uptake, indicating that GABA transport might be regulated by opioid receptors. For further investigation we have expressed GAT1 of mouse brain together with mouse δOR and μOR in Xenopus oocytes. The function of GAT1 was analyzed in terms of Na(+)-dependent [(3)H]GABA uptake as well as GAT1-mediated currents. Coexpression of δOR led to reduced number of fully functional GAT1 transporters, reduced substrate translocation, and GAT1-mediated current. Activation of δOR further reduced the rate of GABA uptake as well as GAT1-mediated current. Coexpression of μOR, as well as μOR activation, affected neither the number of transporters, nor rate of GABA uptake, nor GAT1-mediated current. Inhibition of GAT1-mediated current by activation of δOR was confirmed in whole-cell patch-clamp experiments on rat brain slices of periaqueductal gray. We conclude that inhibition of GAT1 function will strengthen the inhibitory action of the GABAergic system and hence may contribute to acupuncture-induced analgesia.
Figure 1. Effect of chronic morphine treatment on GABA uptake activity in rats. Uptake activity was measured in synaptosomes of untreated rats (control) and of rats after 10 days of morphine injection twice a day (hatched bars). Data were normalized to the rate of uptake into synaptosomes of controls. For treated rats, rate of GABA uptake was determined 12 hr after the termination of treatment (morphine), with an additional injection of morphine (s.c. 10 mg/kg) 1 hr before sacrificing (morphine + morphine) or with an additional injection of both morphine and naloxone (i.p. 2 mg/kg) 1 hr before sacrificing (morphine + morphine + naloxone). *P < 0.05, **P < 0.01 compared to data from control animals; n = 5 in each group. Error bars represent SEM.
Figure 2. Dependence of GAT1-mediated rate of [3H]GABA uptake on different amounts of coinjected cRNA. For cRNA of δOR (filled squares) or rα2β pumps (filled circles), the amount of injected cRNA for GAT1 was 40 ng. Data were normalized for each batch of oocytes to the respective value obtained from oocytes not coinjected with cRNA for δOR or rα2β and are presented as means ± SEM from 2 to 3 batches of oocytes (with 8–10 oocytes per batch). The dependence of rate of GABA uptake on δOR-cRNA was fitted by (1).
Figure 3. Effect of opioid receptor coexpression on GAT1-mediated rate of GABA uptake. (a) 40 ng of GAT1 cRNA alone or 40 ng of GAT1 and 20 ng of cRNA for δOR were injected into oocytes. Application of 100 nM naloxone gave slight recovery of GAT1 inhibition by the coexpressed δOR. (b) 40 ng of GAT1 cRNA alone or 40 ng of GAT1 and 10 ng of cRNA for δOR or μOR were injected into oocytes. The unspecific rate of uptake was subtracted from the uptake rate of injected oocytes. δOR was activated by application of 100 nM DPDPE, μOR by 100 nM DAMGO. Data were normalized for each batch of oocytes to the respective value obtained from oocytes not coinjected with cRNA for δOR and are presented as means of rates of [3H]GABA uptake ± SEM from 2 to 4 batches of oocytes (with 5–10 oocytes per batch), *P < 0.05.
Figure 4. Effects of opioid receptor coexpression on GAT1-mediated current. (a) Current traces in response to rectangular voltage pulses before, during, and after application of 100 μM GABA to oocytes with expressed GAT1. Effects on the voltage dependence of steady-state GAT1-mediated currents in oocytes coexpressed with δOR or μOR (10 ng cRNA coinjected) (b) and of their activation by 100 nM DPDPE or DAMGO, respectively (c). Data in (b) and (c) represent averages ± SEM of at least 6 oocytes.
Figure 5. Effects of δOR coexpression and its activation by 100 nM DPDPE on GAT1-mediated transient charge movements. 40 ng of GAT1 cRNA alone or 40 ng of GAT1 and 20 ng of δOR cRNA were injected into oocytes. (a) Voltage dependencies of moved charge in response to rectangular potential jumps were obtained by integration of the respective transient currents and were fitted by (2) (see lines). (b) The rate constants were obtained by fitting an exponential to the transient current signal, and the voltage dependencies were fitted by (3) (see lines). Data represent means ± SEM; n = 6-7 oocytes for each group.
Figure 6. GAT1 expression in Xenopus oocyte membrane fractions. (a) Lane 1: noninjected oocytes, lane 2: oocytes injected with 40 ng of GAT1 cRNA and 20 ng of δOR, and lane 3: oocytes injected with 40 ng of GAT1 cRNA alone. The bands at about 60 kDa (see arrow) represent GAT1 monomers. (b) GAT1 monomer band intensities averaged from 5 batches of oocytes and normalized to the respective batch injected with cRNA for GAT1 only.
Figure 7. Effect of δOR activation on GAT1-mediated current in rat brain slices from PAG area. Whole-cell patch-clamp recordings were performed on PAG neurons superfusing with different solutions. (a) Steady-state current voltage dependencies of a single experiment with solution sequence as given in the text. The inset shows the GAT1-mediated current determined as tiagabine-sensitive steady-state current. Open squares represent current in the absence and filled squares in the presence of 100 nM DPDPE. (b) GAT1-mediated current from 5 experiments. Data were normalized to the current at −100 mV in the absence of DPDPE and represent averages (±SEM). The value of “one” corresponds to 41.4 ± 20.0 pA.
Barbaresi,
GABA transporter-1 (GAT-1) immunoreactivity in the cat periaqueductal gray matter.
1998, Pubmed
Barbaresi,
GABA transporter-1 (GAT-1) immunoreactivity in the cat periaqueductal gray matter.
1998,
Pubmed
Beckman,
Neurotransmitter transporters: regulators of function and functional regulation.
1998,
Pubmed
Beckman,
Protein kinase C regulates the interaction between a GABA transporter and syntaxin 1A.
1998,
Pubmed
Bicho,
Rapid substrate-induced charge movements of the GABA transporter GAT1.
2005,
Pubmed
Bie,
Trafficking of central opioid receptors and descending pain inhibition.
2007,
Pubmed
Borden,
GABA transporter heterogeneity: pharmacology and cellular localization.
1996,
Pubmed
Cahill,
Up-regulation and trafficking of delta opioid receptor in a model of chronic inflammation: implications for pain control.
2003,
Pubmed
Chao,
Ionic storm in hypoxic/ischemic stress: can opioid receptors subside it?
2010,
Pubmed
Commons,
Translocation of presynaptic delta opioid receptors in the ventrolateral periaqueductal gray after swim stress.
2003,
Pubmed
Corey,
Protein kinase C modulates the activity of a cloned gamma-aminobutyric acid transporter expressed in Xenopus oocytes via regulated subcellular redistribution of the transporter.
1994,
Pubmed
,
Xenbase
Costantino,
Opioid receptor heteromers in analgesia.
2012,
Pubmed
Deken,
Transport rates of GABA transporters: regulation by the N-terminal domain and syntaxin 1A.
2000,
Pubmed
,
Xenbase
Deng,
Interactions of Na+,K+-ATPase and co-expressed delta-opioid receptor.
2009,
Pubmed
,
Xenbase
Eisinger,
Chronic morphine treatment inhibits opioid receptor desensitization and internalization.
2002,
Pubmed
El Far,
G-protein-coupled receptors for neurotransmitter amino acids: C-terminal tails, crowded signalosomes.
2002,
Pubmed
Erbs,
Distribution of delta opioid receptor-expressing neurons in the mouse hippocampus.
2012,
Pubmed
Fields,
State-dependent opioid control of pain.
2004,
Pubmed
Gong,
GABA transporter-1 activity modulates hippocampal theta oscillation and theta burst stimulation-induced long-term potentiation.
2009,
Pubmed
Han,
Acupuncture and endorphins.
2004,
Pubmed
He,
Differential synaptic localization of the glutamate transporter EAAC1 and glutamate receptor subunit GluR2 in the rat hippocampus.
2000,
Pubmed
Heijna,
Opioid receptor-mediated inhibition of 3H-dopamine and 14C-acetylcholine release from rat nucleus accumbens slices. A study on the possible involvement of K+ channels and adenylate cyclase.
1992,
Pubmed
Hilgemann,
GAT1 (GABA:Na+:Cl-) cotransport function. Database reconstruction with an alternating access model.
1999,
Pubmed
,
Xenbase
Hu,
Hyperalgesic effects of gamma-aminobutyric acid transporter I in mice.
2003,
Pubmed
Illes,
Modulation of transmitter and hormone release by multiple neuronal opioid receptors.
1989,
Pubmed
Jasmin,
GABA puts a stop to pain.
2004,
Pubmed
Kang,
delta-Opioid receptors protect from anoxic disruption of Na+ homeostasis via Na+ channel regulation.
2009,
Pubmed
,
Xenbase
Kavanaugh,
Electrogenic uptake of gamma-aminobutyric acid by a cloned transporter expressed in Xenopus oocytes.
1992,
Pubmed
,
Xenbase
Keynan,
gamma-Aminobutyric acid transport in reconstituted preparations from rat brain: coupled sodium and chloride fluxes.
1988,
Pubmed
Knabl,
Reversal of pathological pain through specific spinal GABAA receptor subtypes.
2008,
Pubmed
Krause,
Identification and selective inhibition of the channel mode of the neuronal GABA transporter 1.
2005,
Pubmed
,
Xenbase
Lafaire,
Voltage dependence of the rheogenic Na+/K+ ATPase in the membrane of oocytes of Xenopus laevis.
1986,
Pubmed
,
Xenbase
Lee,
Dopamine transporter cell surface localization facilitated by a direct interaction with the dopamine D2 receptor.
2007,
Pubmed
Levenson,
Long-term potentiation and contextual fear conditioning increase neuronal glutamate uptake.
2002,
Pubmed
Liu,
Effect of mutation of glycosylation sites on the Na+ dependence of steady-state and transient currents generated by the neuronal GABA transporter.
1998,
Pubmed
,
Xenbase
Loo,
Role of Cl- in electrogenic Na+-coupled cotransporters GAT1 and SGLT1.
2000,
Pubmed
,
Xenbase
Lou,
Modulation of protein kinase C and cAMP-dependent protein kinase by delta-opioid.
1997,
Pubmed
Lou,
Nociceptin/orphanin FQ activates protein kinase C, and this effect is mediated through phospholipase C/Ca2+ pathway.
1997,
Pubmed
Mager,
Steady states, charge movements, and rates for a cloned GABA transporter expressed in Xenopus oocytes.
1993,
Pubmed
,
Xenbase
Mager,
Ion binding and permeation at the GABA transporter GAT1.
1996,
Pubmed
,
Xenbase
Malan,
Spinal GABA(A) and GABA(B) receptor pharmacology in a rat model of neuropathic pain.
2002,
Pubmed
Mansour,
Opioid-receptor mRNA expression in the rat CNS: anatomical and functional implications.
1995,
Pubmed
Marazziti,
GPR37 associates with the dopamine transporter to modulate dopamine uptake and behavioral responses to dopaminergic drugs.
2007,
Pubmed
Matthes,
Loss of morphine-induced analgesia, reward effect and withdrawal symptoms in mice lacking the mu-opioid-receptor gene.
1996,
Pubmed
Mayer,
Antagonism of acupuncture analgesia in man by the narcotic antagonist naloxone.
1977,
Pubmed
Miyamae,
Delta opioid receptor mediates phospholipase C activation via Gi in Xenopus oocytes.
1993,
Pubmed
,
Xenbase
Moore,
Partial peripheral nerve injury promotes a selective loss of GABAergic inhibition in the superficial dorsal horn of the spinal cord.
2002,
Pubmed
Ogimoto,
G protein-coupled receptors regulate Na+,K+-ATPase activity and endocytosis by modulating the recruitment of adaptor protein 2 and clathrin.
2000,
Pubmed
Oguri,
Enhanced binding of morphine and nalorphine to opioid delta receptor by glucuronate and sulfate conjugations at the 6-position.
1987,
Pubmed
Ortiz,
Extracellular signal-regulated protein kinases (ERKs) and ERK kinase (MEK) in brain: regional distribution and regulation by chronic morphine.
1995,
Pubmed
Pomeranz,
Naloxone blockade of acupuncture analgesia: endorphin implicated.
1976,
Pubmed
Pu,
Hippocampal long-term potentiation is reduced by chronic opiate treatment and can be restored by re-exposure to opiates.
2002,
Pubmed
Quick,
Second messengers, trafficking-related proteins, and amino acid residues that contribute to the functional regulation of the rat brain GABA transporter GAT1.
1997,
Pubmed
,
Xenbase
Risso,
Sodium-dependent GABA-induced currents in GAT1-transfected HeLa cells.
1996,
Pubmed
Rothstein,
Localization of neuronal and glial glutamate transporters.
1994,
Pubmed
Schloss,
Neurotransmitter transporters. A novel family of integral plasma membrane proteins.
1992,
Pubmed
Schmalzing,
Up-regulation of sodium pump activity in Xenopus laevis oocytes by expression of heterologous beta 1 subunits of the sodium pump.
1991,
Pubmed
,
Xenbase
Schoffelmeer,
Synergistically interacting dopamine D1 and NMDA receptors mediate nonvesicular transporter-dependent GABA release from rat striatal medium spiny neurons.
2000,
Pubmed
Shenoy,
Multifaceted roles of beta-arrestins in the regulation of seven-membrane-spanning receptor trafficking and signalling.
2003,
Pubmed
Trujillo,
Inhibition of morphine tolerance and dependence by the NMDA receptor antagonist MK-801.
1991,
Pubmed
Ullensvang,
Differential developmental expression of the two rat brain glutamate transporter proteins GLAST and GLT.
1997,
Pubmed
Vasilets,
Voltage-dependent inhibition of the sodium pump by external sodium: species differences and possible role of the N-terminus of the alpha-subunit.
1993,
Pubmed
,
Xenbase
Xia,
Regulation of the glutamate transporter EAAC1 by expression and activation of delta-opioid receptor.
2006,
Pubmed
,
Xenbase
Xu,
Morphine withdrawal increases glutamate uptake and surface expression of glutamate transporter GLT1 at hippocampal synapses.
2003,
Pubmed
Zhang,
Neuroprotective role of delta-opioid receptors in cortical neurons.
2002,
Pubmed
Zhang,
Excitatory amino acid receptor antagonists and electroacupuncture synergetically inhibit carrageenan-induced behavioral hyperalgesia and spinal fos expression in rats.
2002,
Pubmed
Zhang,
Role of delivery and trafficking of delta-opioid peptide receptors in opioid analgesia and tolerance.
2006,
Pubmed
Zhu,
Different functional roles of arginine residues 39 and 61 and tyrosine residue 98 in transport and channel mode of the glutamate transporter EAAC1.
2004,
Pubmed
,
Xenbase
Zhu,
Expression and transport function of the glutamate transporter EAAC1 in Xenopus oocytes is regulated by syntaxin 1A.
2005,
Pubmed
,
Xenbase
