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Differential trafficking of adenosine receptors in hippocampal neurons monitored using GFP- and super-ecliptic pHluorin-tagged receptors
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Baines, A. E., Corrêa, Sônia A. L., Irving, A. J. and Frenguelli, Bruno G. (2011) Differential trafficking of adenosine receptors in hippocampal neurons monitored using GFP- and super-ecliptic pHluorin-tagged receptors. Neuropharmacology, Vol.61 (No.1-2). pp. 1-11. doi:10.1016/j.neuropharm.2011.02.005 ISSN 0028-3908.
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Official URL: http://dx.doi.org/10.1016/j.neuropharm.2011.02.005
Abstract
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.
| Item Type: | Journal Article | ||||
|---|---|---|---|---|---|
| Subjects: | Q Science > QH Natural history > QH301 Biology Q Science > QP Physiology |
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| Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) | ||||
| Library of Congress Subject Headings (LCSH): | Adenosine -- Receptors, Adenosine -- Receptors -- Physiological transport, G proteins, Neurons | ||||
| Journal or Publication Title: | Neuropharmacology | ||||
| Publisher: | Elsevier | ||||
| ISSN: | 0028-3908 | ||||
| Official Date: | July 2011 | ||||
| Dates: |
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| Volume: | Vol.61 | ||||
| Number: | No.1-2 | ||||
| Page Range: | pp. 1-11 | ||||
| DOI: | 10.1016/j.neuropharm.2011.02.005 | ||||
| Status: | Peer Reviewed | ||||
| Publication Status: | Published | ||||
| Access rights to Published version: | Restricted or Subscription Access | ||||
| Funder: | Medical Research Council (Great Britain) (MRC) |
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