Neurotransporter Atlas: GLT1

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About

Glutamate is the major excitatory transmitter in the central nervous system (Danbolt, Prog. Neurobiol. 65:1-105, 2001), and it is inactivated by cellular uptake, mostly catalyzed by the glutamate transporters GLT1 (slc1a2, excitatory amino acid transporter [EAAT2]) subtype expressed at high levels in brain astrocytes and at lower levels in neurons. Three C-terminal variants of EAAT2 exist: GLT1a (Pines et al., Nature 360:464-467, 1992), GLT1b (Utsunomiya-Tate et al., FEBS Lett 416:312-326,1997), and GLT1c (Rauen et al., Neurochem. Int. 45:1095-1106, 2004). The Neurotransporter Atlas: GLT1 is an interactive resource providing access to a comprehensive collection of microscopic images showing the brain-wide distribution of GLT1 in the mouse and rat brain, visualized by immunohistochemistry using antibodies against GLT1a and GLT1b. To facilitate identification of anatomical location adjacent section were stained to reveal cyto- and myeloarchitecture. 

Access image repository

The virtual microscopy viewer allows interactive zooming and panning. Original images are available for download via separate link.

Re-use of data from this repository is allowed provided that reference is given to the following publication: Holmseth S, Scott HA, Real K, Lehre KP, Leergaard TB, Bjaalie JG, Danbolt NC (2009) The concentrations and distributions of three C-terminal variants of the GLT1 (EAAT2; slc1a2) glutamate transporter protein in rat brain tissue suggest differential regulation. Neuroscience 162:1055-71; doi:10.1016/j.neuroscience.2009.03.048 

Case # Species Orientation Staining Image repository Download
NM01 Mouse Coronal GLT1a Filmstrip viewer Tiffs
NM01 Mouse Coronal GLT1b Filmstrip viewer Tiffs
NM01 Mouse Coronal Myelin Filmstrip viewer Tiffs
NM01 Mouse Coronal Thionin Filmstrip viewer Tiffs
R4372 Rat Sagittal GLT1a Filmstrip viewer Tiffs
R4372 Rat Sagittal GLT1b Filmstrip viewer Tiffs
R4372 Rat Sagittal Thionin Filmstrip viewer Tiffs

Experimental procedures in brief

Adult animals (C57bl6 mouse, Wistar rat) were transcardially perfused with 4 % formaldehyde and 0.2 % glutaraldehyde.

The mouse brain was cut coronally at 40 μm on a freezing microtome (mouse). Free floating sections were treated with 1M ethanolamine–HCl (pH 7.4), blocked with 10% newborn calf serum TBS (300 mM NaCl and 100 mM Tris–HCl pH 7.4) with 0.25% Triton X-100, and incubated overnight with primary antibodies diluted in blocking solution, followed by biotinylated secondary antibodies also diluted in blocking solution and developed with the biotin–streptavidin–peroxidase system and diaminobenzidine. The GLT1a antibody (Ab#355) was used at 0.06 μg/ml and the GLT1b antibody (Ab#357) at 0.08 μg/ml. 

The rat brain was cut sagitally at 40 μm on a vibratome at room temperature. Free floating sections were treated with 1M ethanolamine–HCl (pH 7.4), blocked with 10% newborn calf serum TBS (300 mM NaCl and 100 mM Tris–HCl pH 7.4), and incubated overnight with primary antibodies diluted in blocking solution, followed by biotinylated secondary antibodies also diluted in blocking solution and developed with the biotin–streptavidin–peroxidase system and diaminobenzidine as described. The GLT1a antibody (Ab#355) was used at 0.3 μg/ml and the GLT1b antibody (Ab#357) at 3 μg/ml.

Neighboring sections were counterstained with a standard thionine stain or with a combined stain for myelin (according to Woelche, 1942) and cytoarchitecture (standard Cresyl Violet counterstain). 

Bregma levels were determined using a standard stereotaxic atlas of the mouse (Franklin and Paxinos, The mouse brain in stereotaxic coordinates, Elsevier, 2001) or rat (Paxinos and Watson, The rat brain in stereotaxic coordinates, Elsevier 2008) brain. For further details, see Holmseth et al. (Neuroscience 162:1055-71, 2009) and http://www.neurotransporter.org/

Contributing laboratories

The Neuro Transporter Group (http://www.neurotransporter.org), Centre for Molecular Biology and Neuroscience & Institute of Basic Medical Sciences, Department of Anatomy, University of Oslo, P.O. Box 1105 Blindern, N - 0317 Oslo, Norway: Experimental work, immunohistochemistry. People: Silvia Holmseth, Henriette Danbolt, Knut P. Lehre, Niels C. Danbolt

Neural Systems Laboratory (http://www.nesys.uio.no), Centre for Molecular Biology and Neuroscience & Institute of Basic Medical Sciences, Department of Anatomy, University of Oslo, P.O. Box 1105 Blindern, N - 0317 Oslo, Norway: Histological processing, image acquisition, atlas repository. People: Jan O. Kjøde, Ivar A. Moene, Dmitri Darine, Saurabh Jain, Anna T. Bore, Trygve B. Leergaard, Jan G. Bjaalie

Funded by:

  • The neuroinformatics components of this resource have been funded by the Human Brain Project through the European Union Seventh Framework Program (FP7/2007-2013) under grant agreement no. 604102 (HBP)

Contact

j.g.bjaalie@medisin.uio.no