Difference between revisions of "Neurotransporter Atlas: GLT1"

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'''Welcome to NT-atlas, a part of the Rodent Brain WorkBench'''
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== <span style="font-family:georgia,serif;">About</span> ==
  
''NT-atlas''&nbsp;is an online public neuroscience data repository for extensive documentation of the distribution of glutamate transporters (Excitatory Amino Acid Transporters: EAATs). It provides access to collections of high resolution image data showing the distribution of glutamate receptors in mouse and rat brains.
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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. 
  
''NT-atlas''&nbsp;is part of the Rodent Brain WorkBench, a new research and development project funded by The Research Council of Norway and the Centre for Molecular Biology and Neuroscience.
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== <span style="font-family:georgia,serif;">Access image repository</span> ==
  
This website is under development. The first data presented concern distributions of the EAAT2 (GLT1) subtype and are based on data published in:
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The virtual microscopy viewer allows interactive zooming and panning. Original images are available for download via separate link.
  
<span style="font-family: Arial, Helvetica, sans-serif; font-size: 11.960000038147px; line-height: 18.3944797515869px; text-indent: -35px;">Holmseth, Silvia; Scott, Heather A; Real, Katia; Lehre, Knut Petter Dæhlin; Leergaard, Trygve Brauns; Bjaalie, Jan G. & Danbolt, Niels Christian&nbsp;(2009).&nbsp;The concentrations and distributions of three C-terminal variants of the GLT1 (EAAT2; slc1a2) glutamate transporter protein in rat brain tissue suggest differential regulation.&nbsp;</span>''Neuroscience''<span style="font-family: Arial, Helvetica, sans-serif; font-size: 11.960000038147px; line-height: 18.3944797515869px; text-indent: -35px;">. &nbsp;ISSN&nbsp;0306-4522.&nbsp;</span>''&nbsp;162''<span style="font-family: Arial, Helvetica, sans-serif; font-size: 11.960000038147px; line-height: 18.3944797515869px; text-indent: -35px;">(4),&nbsp;s&nbsp;1055-&nbsp;1071 .&nbsp;doi:</span>[http://dx.doi.org/10.1016/j.neuroscience.2009.03.048 10.1016/j.neuroscience.2009.03.048]
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'''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;<span style="font-family:georgia,serif;"><span style="font-size: 11.960000038147px; line-height: 18.3944797515869px; text-indent: -35px;">&nbsp;doi:</span>[http://dx.doi.org/10.1016/j.neuroscience.2009.03.048 10.1016/j.neuroscience.2009.03.048]</span> 
  
Use the links on this page to access the repository. Selected images can be viewed side-by-side in a viewer tool.&nbsp;
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{| class="wikitable"
 
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!Case #
 
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!Species
 
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!Orientation
== Goals and background ==
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!Staining
 
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!Image repository
'''The primary&nbsp;goal&nbsp;of&nbsp;the NT-atlas project is&nbsp;:'''
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!Download
 
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|-
*To better understand the system of neurotransporters by mapping the distributions of the glutamate transporters across the entire mouse and rat brain
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|NM01
 
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|Mouse
<br/>'''NT-atlas is a part of the Rodent Brain WorkBench project. The goals of this project are:'''
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|Coronal
 
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|GLT1a
*to construct a comprehensive database, or several interoperable database systems, that will include structure and structure-function data from whole brain and selected brain regions
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|[http://cmbn-navigator.uio.no/navigator/filmstrip_viewer.html?publicOnly=true&entityType=block&entityId=2568&selectIndex=9&metadataKeyName=Bregma%20Level# Filmstrip viewer]
*to develop tools for visualization and analysis linked to the databases and allowing detailed investigations to be performed on the data
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|[http://cmbn-navigator.uio.no/navigator/feeder/all_originals/?id=2568 Tiffs]
*to generate a digital atlas system for multiple categories of image data from rat and mouse brain
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|-
 
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|NM01
&nbsp;
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|Mouse
 
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|Coronal
'''Background:'''
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|GLT1b
 
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|[http://cmbn-navigator.uio.no/navigator/filmstrip_viewer.html?publicOnly=true&entityType=block&entityId=2567&selectIndex=9&metadataKeyName=Bregma%20Level# Filmstrip viewer]
Glutamate is the major excitatory transmitter in the central nervous system, and is inactivated by cellular uptake catalyzed by the glutamate transporters (or Excitatory Amino Acid Transporters, EAATs): EAAT1 (GLAST), (EAAT2 (GLT1), EAAT3 (EAAC), EAAT4 and EAAT5. The EAATs belong to Solute Carrier Family (slc) number 1 (for review and references, see, Danbolt, 2001).
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|[http://cmbn-navigator.uio.no/navigator/feeder/all_originals/?id=2567 Tiffs]
 
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|-
'''EAAT2 (slc1a2)&nbsp;'''is the quantitatively and functionally most important of the five EAAT subtypes in the mature brain. It accounts for more than 90&nbsp;% of the total glutamate uptake activity in the forebrain and is essential for normal brain function (Danbolt et al., 1992; Haugeto et al., 1996; Tanaka et al., 1997). EAAT2 protein is expressed in astrocytes in the normal and mature nervous system (Levy et al., 1993; Chaudhry et al., 1995; Lehre et al., 1995).
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|NM01
 
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|Mouse
Although it has been clear that EAAT2 protein is expressed in neurons in the normal and mature brain, it has been equally clear that EAAT2 mRNA is present in both neurons and astroglia (Torp et al., 1994, 1997; Berger and Hediger, 1998). It now turns out that EAAT2 protein is expressed in synaptic terminals, but at 10 times lower densities than in astroglia, and that the glutamate uptake into terminals is fully dependent on the EAAT2 gene (Furness et al. 2008).
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|Coronal
 
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|Myelin
EAAT2 is variably spliced in many different ways and is found with alternative N-termini or C-termini, as well as with other modifications (for references see: Meyer et al., 1999; Danbolt, 2001). Three C-terminal variants of EAAT2 exist:
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|[http://cmbn-navigator.uio.no/navigator/filmstrip_viewer.html?publicOnly=true&entityType=block&entityId=2565&selectIndex=12&metadataKeyName=Bregma%20Level# Filmstrip viewer]
 
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|[http://cmbn-navigator.uio.no/navigator/feeder/all_originals/?id=2565 Tiffs]
*GLT1a (Pines et al., 1992)
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|-
*GLT1b (Utsunomiya-Tate et al., 1997)
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|NM01
*GLT1c (Rauen et al., 2004) &nbsp;
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|Mouse
 
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|Coronal
&nbsp;
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|Thionin
 
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|[http://cmbn-navigator.uio.no/navigator/filmstrip_viewer.html?publicOnly=true&entityType=block&entityId=2566&selectIndex=11&metadataKeyName=Bregma%20Level# Filmstrip viewer]
The major neuronal mRNA isoform is GLT1a (Berger et al., 2005) and GLT1a is the only form found in terminals (Furness et al. 2008; Holmseth et al., 2009). The question of whether GLT1b is present in neurons or not has been controversial (for references and discussion see: Holmseth et al., 2009).
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|[http://cmbn-navigator.uio.no/navigator/feeder/all_originals/?id=2566 Tiffs]
 
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|-
 
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|R4372
 
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|Rat
'''The repository contains now two series of sections:'''
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|Sagittal
 
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|GLT1a
*'''NM01:&nbsp;'''Mouse cryo-sections processed according to Procedure #1 (see Experimental procedures) with detergent (to maximize tissue penetation). This reveal regional differences in expression levels, but comes at the expense of cellular details.
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|[http://cmbn-navigator.uio.no/navigator/filmstrip_viewer.html?publicOnly=true&entityType=block&entityId=2561&selectIndex=6&metadataKeyName=Bregma%20Level# Filmstrip viewer]
*'''R4372:&nbsp;'''Rat vibratome sections processed according to Procedure #2 (see Experimental procedures) as free-floating sections that have neither been frozen nor exposed to detergents. This procedure is best for visualizing cellular details.
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|[http://cmbn-navigator.uio.no/navigator/feeder/all_originals/?id=2561 Tiffs]
 
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|-
&nbsp;For more information, see: [http://www.neurotransporter.org/ http://www.neurotransporter.org/]
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|R4372
 
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|Rat
== Experimental procedures ==
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|Sagittal
 
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|GLT1b
For methodological details, see Holmseth and coworkers (2009). Animals were perfusion fixed with 4&nbsp;% formaldehyde and 0.2&nbsp;% glutaraldehyde in 0.1 M sodium phosphate buffer.
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|[http://cmbn-navigator.uio.no/navigator/filmstrip_viewer.html?publicOnly=true&entityType=block&entityId=2563&selectIndex=5&metadataKeyName=Bregma%20Level# Filmstrip viewer]
 
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|[http://cmbn-navigator.uio.no/navigator/feeder/all_originals/?id=2563 Tiffs]
'''Procedure 1:&nbsp;'''Sections were cut from frozen tissue and processed with Triton X-100. Free floating sections (40 μm thick) 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 as described (Danbolt et al., 1998). The GLT1a antibody (Ab#355) was used at 0.06 μg/ml and the GLT1b antibody (Ab#357) at 0.08 μg/ml.&nbsp;
 
 
 
'''Procedure 2:'''&nbsp;Sections were cut on a Vibratome and the tissue was not frozen. Free floating sections (40 μm thick) 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 (Danbolt et al., 1998). The GLT1a antibody (Ab#355) was used at 0.3 μg/ml and the GLT1b antibody (Ab#357) at 3 μg/ml.
 
 
 
&nbsp;
 
 
 
=== General histochemistry ===
 
 
 
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) &nbsp;
 
 
 
&nbsp;
 
 
 
=== Atlas coordinate values ===
 
 
 
For all sections, Bregma-related stereotaxic coordinates (“Bregma” in image repository) were found by anchoring selected sections to atlas diagrams using multiple anatomical landmarks. For the remaining sections, coordinates were interpolated from serial number and section thickness. Nearest matching atlas diagrams (“Atlas bregma” and corresponding “Atlas interaural” in image repository) were determined from closest matching bregma values in Franklin and Paxinos (2007, mouse brain atlas) and Paxinos and Watson (2008, rat brain atlas), respectively. &nbsp;
 
 
 
 
 
 
 
== Access image repository ==
 
 
 
The present &nbsp;viewing tool allows the user to navigate within and across pairs of coronal or sagittal section images obtained from normal mice and rats.&nbsp;The EAAT atlas project is currently exploring different approaches for viewing of microscopic images via the web. The viewing tool is used for rapid inspection of large series of sections with images of higher resolution.&nbsp;
 
 
 
Re-use of data from this repository is allowed provided that reference is given to the following publication:
 
 
 
<span style="font-family: Arial, Helvetica, sans-serif; font-size: 11.960000038147px; line-height: 18.3944797515869px; text-indent: -35px;">Holmseth, Silvia; Scott, Heather A; Real, Katia; Lehre, Knut Petter Dæhlin; Leergaard, Trygve Brauns; Bjaalie, Jan G. & Danbolt, Niels Christian&nbsp;(2009).&nbsp;The concentrations and distributions of three C-terminal variants of the GLT1 (EAAT2; slc1a2) glutamate transporter protein in rat brain tissue suggest differential regulation.&nbsp;</span>''Neuroscience''<span style="font-family: Arial, Helvetica, sans-serif; font-size: 11.960000038147px; line-height: 18.3944797515869px; text-indent: -35px;">. &nbsp;ISSN&nbsp;0306-4522.&nbsp;</span>''&nbsp;162''<span style="font-family: Arial, Helvetica, sans-serif; font-size: 11.960000038147px; line-height: 18.3944797515869px; text-indent: -35px;">(4),&nbsp;s&nbsp;1055-&nbsp;1071 .&nbsp;doi:</span>[http://dx.doi.org/10.1016/j.neuroscience.2009.03.048 10.1016/j.neuroscience.2009.03.048]
 
 
 
 
 
 
 
{| border="1" width="100%" style="font-family: Verdana, Arial, Helvetica, sans-serif;"
 
|- style="color: rgb(0, 0, 0);"
 
! Name
 
! Species
 
! CuttingDirection
 
! BlckDate
 
! Comments
 
|- style="color: rgb(0, 0, 0);"
 
| NM01&nbsp;
 
| MOUSE&nbsp;
 
| CORONAL&nbsp;
 
| 2006-07-07&nbsp;
 
| GLT1a, GLT1b, thionin, myelin&nbsp;
 
|- style="color: rgb(0, 0, 0);"
 
| R4372&nbsp;
 
| RAT&nbsp;
 
| SAGITTAL&nbsp;
 
| 2009-01-07&nbsp;
 
| thionin,GLT1a,GLT1b, Right hemisphere&nbsp;
 
|}
 
 
 
<br/>Below, selected images are available for inspection.<br/><br/>To view the high resolution images, choose from the table above.
 
 
 
 
 
 
 
{| style="width: 1000px" border="0" cellpadding="0" cellspacing="0"
 
 
|-
 
|-
| [[File:Glt slide36 thumb.jpg|thumb|left|350px|A: R4372s036_GLT1A]]<br/>
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|R4372
| [[File:Glt slide37 thumb.jpg|thumb|left|350px|B: R4372s037_GLT1B]]<br/>
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|Rat
| [[File:Glt slide46 thumb.jpg|thumb|left|350px|B: R4372s046_GLT1A]]<br/>
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|Sagittal
| [[File:Glt slide47 thumb.jpg|thumb|left|350px|B: R4372s047_GLT1B]]<br/>
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|Thionin
 +
|[http://cmbn-navigator.uio.no/navigator/filmstrip_viewer.html?publicOnly=true&entityType=block&entityId=2562&selectIndex=4&metadataKeyName=Bregma%20Level# Filmstrip viewer]
 +
|[http://cmbn-navigator.uio.no/navigator/feeder/all_originals/?id=2562 Tiffs]
 
|}
 
|}
 +
== <span style="font-family:georgia,serif;">Experimental procedures in brief</span> ==
  
<gallery mode="packed-hover" heights="180">
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Adult animals (C57bl6 mouse, Wistar rat) were transcardially perfused with 4 % formaldehyde and 0.2 % glutaraldehyde.
Glt slide36 thumb.jpg|alt=slide36_GLT1|R4372s036_GLT1A
 
Glt slide37 thumb.jpg|alt=slide37_GLT1|R4372s037_GLT1B
 
Glt slide46 thumb.jpg|alt=slide46_GLT1|R4372s046_GLT1A
 
Glt slide47 thumb.jpg|alt=slide47_GLT1|R4372s047_GLT1B
 
</gallery>
 
 
 
== Contributing laboratories ==
 
 
 
The Neurotransporter Group - Knut Petter Lehre, Niels Chr. Danbolt<br/>Centre for Molecular Biology and Neuroscience &<br/>Institute of Basic Medical Sciences - Anatomy<br/>University of Oslo<br/>P.O. Box 1105 Blindern<br/>N - 0317 Oslo<br/>Norway<br/>[http://www.neurotransporter.org/ http://www.neurotransporter.org]<br/>[http://www.cmbn.no/ http://www.cmbn.no]
 
 
 
 
 
 
 
Neural Systems and Graphics Computing Laboratory - Trygve B. Leergaard, Jan G. Bjaalie<br/>Centre for Molecular Biology and Neuroscience &<br/>Institute of Basic Medical Sciences - Anatomy<br/>University of Oslo<br/>P.O. Box 1105 Blindern<br/>N - 0317 Oslo<br/>Norway<br/>[http://www.nesys.uio.no/ http://www.nesys.uio.no]<br/>[http://www.rbwb.org/ http://www.rbwb.org]
 
 
 
 
 
 
 
== Credits ==
 
 
 
'''Histology'''<br/><br/>Anna Torbjørg Bore<br/>Saurabh Jain
 
 
 
 
 
 
 
'''Immunohistochemistry'''<br/><br/>Silvia Holmseth<br/>Henriette Danbolt<br/>Knut Petter Lehre<br/>Niels Christian Danbolt
 
 
 
 
 
 
 
'''NT-atlas web-application development team:'''<br/><br/>Jan Olav Kjøde<br/>Ivar Andre Moene&nbsp;<br/>Dmitri Darine&nbsp;<br/>Saurabh Jain<br/>Trygve B. Leergaard<br/>Jan G. Bjaalie
 
 
 
 
 
 
 
== References ==
 
 
 
'''Berger UV, Hediger MA (1998)'''<br/>''Comparative analysis of glutamate transporter expression in rat brain using differential double in situ hybridization.''<br/>Anat Embryol (Berl) 198:13-30.
 
 
 
'''Berger UV, Desilva TM, Chen WZ, Rosenberg PA (2005)&nbsp;'''<br/>''Cellular and subcellular mRNA localization of glutamate transporter isoforms GLT1a and GLT1b in rat brain by in situ hybridization.''<br/>J Comp Neurol 492:78- 89.
 
 
 
'''Chaudhry FA, Lehre KP, Campagne MV, Ottersen OP, Danbolt NC, Storm-Mathisen J (1995)'''<br/>''Glutamate transporters in glial plasma membranes: highly differentiated localizations revealed by quantitative ultrastructural immunocytochemistry.''<br/>Neuron 15:711-720
 
 
 
'''Danbolt NC (2001)'''<br/>''Glutamate uptake.''<br/>Prog Neurobiol 65: 1-105.
 
 
 
'''Danbolt NC, Storm-Mathisen J, Kanner BI (1992)'''<br/>''An [Na+ + K+]coupled L-glutamate transporter purified from rat brain is located in glial cell processes.''<br/>Neuroscience 51:295-310.
 
 
 
'''Danbolt NC, Lehre KP, Dehnes Y, Chaudhry FA, Levy LM (1998)'''<br/>''Localization of transporters using transporter-specific antibodies.''<br/>Methods Enzymol 296:388-407.
 
 
 
'''Franklin K, Paxinos G (2007)'''<br/>''The mouse brain in stereotaxic coordinates (San Diego, Elsevier Academic Press)''
 
 
 
'''Furness D, Dehnes Y, Akhtar A, Rossi D, Hamann M, Grutle N, Gundersen V, Holmseth S, Lehre K, Ullensvang K, Wojewodzic M, Zhou Y, Attwell D, Danbolt N (2008)&nbsp;'''<br/>''A quantitative assessment of glutamate uptake into hippocampal synaptic terminals and astrocytes: New insights into a neuronal role for excitatory amino acid transporter 2 (EAAT2).''<br/>Neuroscience 157:80-94.
 
 
 
'''Holmseth S, Scott HA, Real K, Lehre KP, Leergaard TB, Bjaalie JG and Danbolt NC (2009)&nbsp;'''<br/>''The concentrations and distributions of three C-terminal variants of the GLT1 (EAAT2; slc1a2) glutamate transporter protein in rat brain tissue suggests differential regulation.''<br/>Neuroscience, 2009; doi:10.1016/j.neuroscience.2009.03.048
 
 
 
'''Haugeto Ø, Ullensvang K, Levy LM, Chaudhry FA, Honoré T, Nielsen M, Lehre KP, Danbolt NC (1996)'''<br/>''Brain glutamate transporter proteins form homomultimers.''<br/>J Biol Chem 271:27715-27722.
 
 
 
'''Lehre KP, Levy LM, Ottersen OP, Storm-Mathisen J, Danbolt NC (1995)'''<br/>''Differential expression of two glial glutamate transporters in the rat brain: quantitative and immunocytochemical observations.''<br/>J Neurosci 15:1835-1853.
 
  
'''Levy LM, Lehre KP, Rolstad B, Danbolt NC (1993)'''<br/>''A monoclonal antibody raised against an [Na+ - K+]coupled L- glutamate transporter purified from rat brain confirms glial cell localization.''<br/>FEBS Lett 317:79-84.
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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. 
  
'''Meyer T, Fromm A, Münch C, Schwalenstöcker B, Fray AE, Ince PG, Stamm S, Gron G, Ludolph AC, Shaw PJ (1999)'''<br/>''The RNA of the glutamate transporter EAAT2 is variably spliced in amyotrophic lateral sclerosis and normal individuals.''<br/>J Neurol Sci 170:45-50.
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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.
  
'''Paxinos G, Watson C (2008)'''<br/>''The rat brain in stereotaxic coordinates (San Diego, Elsevier Academic Press)''
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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)
  
'''Pines G, Danbolt NC, Bjørås M, Zhang Y, Bendahan A, Eide L, Koepsell H, Storm-Mathisen J, Seeberg E, Kanner BI (1992)'''<br/>''Cloning and expression of a rat brain L-glutamate transporter.''<br/>Nature 360:464-467.
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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/<span style="font-family:georgia,serif;">
  
'''Rauen T, Wiessner M, Sullivan R, Lee A, Pow DV (2004)'''<br/>''A new GLT1 splice variant: cloning and immunolocalization of GLT1c in the mammalian retina and brain.&nbsp;''<br/>Neurochem Int 45:1095-1106.
+
== <span style="font-family:georgia,serif;">Contributing laboratories</span> ==
  
'''Tanaka K, Watase K, Manabe T, Yamada K, Watanabe M, Takahashi K, Iwama H, Nishikawa T, Ichihara N, Hori S, Takimoto M, Wada K (1997)'''<br/>''Epilepsy and exacerbation of brain injury in mice lacking the glutamate transporter GLT- 1.''<br/>Science 276:1699-1702.
+
'''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
  
'''Torp R, Danbolt NC, Babaie E, Bjørås M, Seeberg E, Storm-Mathisen J, Ottersen OP (1994)'''<br/>''Differential expression of two glial glutamate transporters in the rat brain: an in situ hybridization study.''<br/>Eur J Neurosci 6:936-942.
+
'''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
  
'''Torp R, Hoover F, Danbolt NC, Storm-Mathisen J, Ottersen OP (1997)'''<br/>''Differential distribution of the glutamate transporters GLT1 and rEAAC1 in rat cerebral cortex and thalamus: an in situ hybridization analysis.''<br/>Anat Embryol (Berl) 195:317- 326.
+
== 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)
  
'''Utsunomiya-Tate N, Endou H, Kanai Y (1997)'''<br/>''Tissue specific variants of glutamate transporter GLT-1.''<br/>FEBS Lett 416:312-316.
+
== <span style="font-family:georgia,serif;">Contact</span> ==
  
'''Woelche M (1942)'''<br/>''Eine neue Methode der Markscheidenfarbung.''<br/>Psychol. Neurol. 51:199-202.
+
j.g.bjaalie@medisin.uio.no

Revision as of 21:20, 17 June 2016

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