Differential trafficking of adenosine receptors in hippocampal neurons monitored using GFP- and super-ecliptic pHluorin-tagged receptors. / Baines, A. E.; Correa, S. A. L.; Irving, A.J.; Frenguelli, B. G.
In: Neuropharmacology, Vol. 61, No. 1-2, 2011, p. 1-11.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Differential trafficking of adenosine receptors in hippocampal neurons monitored using GFP- and super-ecliptic pHluorin-tagged receptors
A1 - Baines,A. E.
A1 - Correa,S. A. L.
A1 - Irving,A.J.
A1 - Frenguelli,B. G.
AU - Baines,A. E.
AU - Correa,S. A. L.
AU - Irving,A.J.
AU - Frenguelli,B. G.
PY - 2011
Y1 - 2011
N2 - <p>Adenosine receptors (ARs) modulate many cellular and systems-level processes in the mammalian CNS. However, little is known about the trafficking of ARs in neurons, despite their importance in controlling seizure activity and in neuroprotection in cerebral ischaemia. To address this we examined the agonist-dependent internalisation of C-terminal GFP-tagged A(1)Rs, A(2A)Rs and A(3)Rs in primary hippocampal neurons. Furthermore, we developed a novel super-ecliptic pHluorin (SEP)-tagged AIR which, via the N-terminal SEP tag, reports the cell-surface expression and trafficking of A(1)Rs in real-time. We demonstrate the differential trafficking of ARs in neurons: A(3)Rs internalise more rapidly than A(1)Rs, with little evidence of appreciable A(2A)R trafficking over the time-course of the experiments. Furthermore, the novel SEP-A(1)R construct revealed the time-course of internalisation and recovery of cell-surface expression to occur within minutes of agonist exposure and removal, respectively. These observations highlight the labile nature of A(1)R and A(3)Rs when expressed at the neuronal plasma membrane. Given the high levels of adenosine in the brain during ischaemia and seizures, internalisation of the inhibitory A(1)R may result in hyperexcitability, increased brain damage and the development of chronic epileptic states. (C) 2011 Elsevier Ltd. All rights reserved.</p>
AB - <p>Adenosine receptors (ARs) modulate many cellular and systems-level processes in the mammalian CNS. However, little is known about the trafficking of ARs in neurons, despite their importance in controlling seizure activity and in neuroprotection in cerebral ischaemia. To address this we examined the agonist-dependent internalisation of C-terminal GFP-tagged A(1)Rs, A(2A)Rs and A(3)Rs in primary hippocampal neurons. Furthermore, we developed a novel super-ecliptic pHluorin (SEP)-tagged AIR which, via the N-terminal SEP tag, reports the cell-surface expression and trafficking of A(1)Rs in real-time. We demonstrate the differential trafficking of ARs in neurons: A(3)Rs internalise more rapidly than A(1)Rs, with little evidence of appreciable A(2A)R trafficking over the time-course of the experiments. Furthermore, the novel SEP-A(1)R construct revealed the time-course of internalisation and recovery of cell-surface expression to occur within minutes of agonist exposure and removal, respectively. These observations highlight the labile nature of A(1)R and A(3)Rs when expressed at the neuronal plasma membrane. Given the high levels of adenosine in the brain during ischaemia and seizures, internalisation of the inhibitory A(1)R may result in hyperexcitability, increased brain damage and the development of chronic epileptic states. (C) 2011 Elsevier Ltd. All rights reserved.</p>
KW - Adenosine receptor
KW - Internalisation
KW - Trafficking
KW - Epilepsy
KW - Seizure
KW - Ischaemia
KW - AGONIST-DEPENDENT PHOSPHORYLATION
KW - TEMPORAL-LOBE EPILEPSY
KW - POSTISCHEMIC RAT-BRAIN
KW - A(2A) RECEPTOR
KW - A(3) RECEPTORS
KW - SYNAPTIC-TRANSMISSION
KW - NERVOUS-SYSTEM
KW - A(1) RECEPTORS
KW - MULTIPLE MECHANISMS
KW - CALCIUM-CHANNELS
U2 - 10.1016/j.neuropharm.2011.02.005
DO - 10.1016/j.neuropharm.2011.02.005
M1 - Article
JO - Neuropharmacology
JF - Neuropharmacology
SN - 0028-3908
IS - 1-2
VL - 61
SP - 1
EP - 11
ER -