title: Deriving the time course of glutamate clearance with a deconvolution analysis of astrocytic transporter currents.
abstract: The highest density of glutamate transporters in the brain is found in astrocytes. Glutamate transporters couple the movement of glutamate across the membrane with the co-transport of 3 Na+ and 1 H+ and the counter-transport of 1 K+. The stoichiometric current generated by the transport process can be monitored with whole-cell patch-clamp recordings from astrocytes. The time course of the recorded current is shaped by the time course of the glutamate concentration profile to which astrocytes are exposed, the kinetics of glutamate transporters, and the passive electrotonic properties of astrocytic membranes. Here we describe the experimental and analytical methods that can be used to record glutamate transporter currents in astrocytes and isolate the time course of glutamate clearance from all other factors that shape the waveform of astrocytic transporter currents. The methods described here can be used to estimate the lifetime of flash-uncaged and synaptically-released glutamate at astrocytic membranes in any region of the central nervous system during health and disease.
authors: Synaptic Physiology Section, National Institute of Neurological Disorders and Stroke, National Institutes of Health, USA. scimemia@ninds.nih.gov
authors: Annalisa
authors: A
authors: Scimemi
authors: Jeffrey S
authors: JS
authors: Diamond
mesh_terms: D027322:Amino Acid Transport System X-AG / Q000378:metabolism*; D000818:Animals; D001253:Astrocytes / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D018698:Glutamic Acid / Q000378:metabolism*; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018408:Patch-Clamp Techniques / Q000379:methods; D011188:Potassium / Q000378:metabolism; D015221:Potassium Channels / Q000378:metabolism; D013569:Synapses / Q000378:metabolism
chemical_list: D027322:Amino Acid Transport System X-AG; D015221:Potassium Channels; D018698:Glutamic Acid; D011188:Potassium
references: Glia. 2012 Feb;60(2):175-88
references: 22052455
references: Neuron. 1995 Sep;15(3):721-8
references: 7546750
references: J Neurosci. 2002 Dec 1;22(23):10153-62
references: 12451116
references: Neuron. 1997 Dec;19(6):1297-308
references: 9427252
references: J Neurosci. 2001 Oct 15;21(20):7969-84
references: 11588170
references: Proc Natl Acad Sci U S A. 2008 Mar 18;105(11):4411-6
references: 18332427
references: Neuroreport. 2001 Apr 17;12(5):1077-80
references: 11303749
references: Nature. 1996 Oct 17;383(6601):634-7
references: 8857541
references: J Neurosci. 2007 Sep 5;27(36):9736-41
references: 17804634
references: Nat Neurosci. 2002 Apr;5(4):325-31
references: 11896395
references: J Neurosci. 1998 Oct 1;18(19):7650-61
references: 9742136
references: J Neurosci. 2009 Nov 18;29(46):14581-95
references: 19923291
references: J Neurosci. 2013 Mar 20;33(12):5312-8
references: 23516295
references: J Neurosci. 2005 Mar 16;25(11):2906-16
references: 15772350
references: J Neurophysiol. 2000 May;83(5):2835-43
references: 10805681
references: J Neurosci. 1998 May 1;18(9):3158-70
references: 9547224
references: Proc Natl Acad Sci U S A. 1996 Apr 30;93(9):4153-8
references: 8633032
references: Glia. 2007 Jan 1;55(1):13-23
references: 17001633
references: J Physiol. 1991 May;436:169-93
references: 1676418
references: J Neurosci. 1999 Aug 15;19(16):6897-906
references: 10436047
references: Prog Neurobiol. 2001 Sep;65(1):1-105
references: 11369436