Difference between revisions of "TTA / TetOff Atlas II (PrP/CamKII)"

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 '''The Tet-Off atlas is an online public neuroscience data repository.'''
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== About ==
 +
The tetracycline-responsive (TetR) gene system is a binary transgenic system in which transgene expression may be activated or silenced by administration of tetracycline, or its derivates (Gossen and Bujard, PNAS 89:5547-5551, 1992; Gossen et al., Science 286:1766-1769, 1995). Such animal models are generated by crossing two different mouse lines, one with a gene modulator (promoter mouse line) and one with the actual disease-causing gene (responder mouse line). In the double transgenic offspring both the regional distribution and intensity of target gene expression depend on the activity of the chosen promoter controlling the expression of a gene activator, which in turn is accessible for experimental modulation. Detailed knowledge about where in the brain gene expression is regulated is essential for the use and interpretation of such models.
  
The ''Tet-Off atlas'' is an interactive resource providing access to brain wide histological images displaying the distribution patterns of promoter activity of the two most popular transgenic tetracycline-responsive (Tet-Off) promoter mouse lines, the Ca2+/calmodulin-dependent protein kinase II (CamKII) and the prion protein (PrP) mouse lines.
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The TetOff atlas 2 (PrP/CamKII) is an interactive resource providing access to a collection of microscopic brain sections displaying the spatial distribution patterns of promoter activity of two popular transgenic tetracycline-responsive (Tet-Off) promoter mouse lines, the prion protein (PrP) mouse line (previously reported in Boy et al., Neuroimage 33:449-462, 2006) and the Ca2+/calmodulin-dependent protein kinase II (CamKII).  
  
The Atlas contains :
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== <span style="font-family:georgia,serif">Access image repository</span> ==
 +
The virtual microscopy viewer allows interactive zooming and panning. Original images are available for download via separate link. Each section has been annotated with names of key landmarks, regions, nuclei, and areas.
  
1.&nbsp;&nbsp;&nbsp; A repository of coronal section images from a CamKII/LacZ mouse, showing the distribution of Tet-Off CamKII promoter activity.
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'''Re-use of data from this repository is allowed provided that reference is given to the following publications:''' 
 +
* CamKII: Odeh F, Leergaard TB, Boy J, Schmidt T, Riess O, Bjaalie JG. Atlas of transgenic Tet-Off Ca2+/calmodulin-dependent protein kinase II and prion protein promoter activity in the mouse brain. NeuroImage 54:2603-2611, 2011 
 +
* PrP: Boy J, Leergaard TB, Schmidt T, Odeh F, Bichelmeier U, Nuber S, Holzmann C, Wree A, Prusiner SB, Bujard HB, Riess O, Bjaalie JG. Expression mapping of tetracycline-responsive prion protein promoter: digital atlasing for generating cell-specific disease-models. NeuroImage 33:449-462, 2006 
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{| class="wikitable"
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!Animal #
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!Genotype
 +
!Image repository
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!Download original
 +
|-
 +
|388.12
 +
|Prnp-tTA:βGal
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|[http://cmbn-navigator.uio.no/navigator/filmstrip_viewer.html?publicOnly=true&entityType=block&entityId=2541&selectIndex=17&metadataKeyName=Bregma%20Level# Filmstrip viewer]
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|[http://cmbn-navigator.uio.no/navigator/feeder/all_originals/?id=2541 Tiffs]
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|-
 +
|317.8
 +
|CamKII-tTA:βGal
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|[http://cmbn-navigator.uio.no/navigator/filmstrip_viewer.html?publicOnly=true&entityType=block&entityId=2542&selectIndex=16&metadataKeyName=Bregma%20Level# Filmstrip viewer]
 +
|[http://cmbn-navigator.uio.no/navigator/feeder/all_originals/?id=2542 Tiffs]
 +
|}
  
2.&nbsp;&nbsp;&nbsp; A repository of coronal section images from a PrP/LacZ mouse, showing the distribution of Tet-Off PrP promoter activity. These data were previously reported in Boy et al. NeuroImage 33:449-462, 2006.
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== <span style="font-family:georgia,serif">Experimental procedures in brief</span> ==
 +
The atlas is based on data from two double transgenic mice, a 40 week old female CamKII/LacZ mouse and a 5 week old female PrP/LacZ mouse (Boy et al., NeuroImage 33: 449-462, 2006). For methodological details concerning gene construct, preparation of histological material, image acquisition, and atlas construction, see Boy et al. (NeuroImage, 33:449-462, 2006), and Odeh et al. (NeuroImage 54:2603-2611, 2011).
  
3.&nbsp;&nbsp;&nbsp; A dual viewer application for side-by-side comparison of spatially corresponding coronal section images from the CamKII/LacZ and PrP/LacZ mouse brains.
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In brief, double-transgenic mice were generated by cross-breeding a CamKII or PrP promoter mouse line with a LacZ responder mouse line. Animals were anaesthetized and transcardially perfused with 4% paraformaledhyde before brains were removed and stained using X-Gal (to label the LacZ product β-galactosidase) and counterstained with Cresyl violet (sections from the CamKII/LacZ brain) or Neutral red (sections from the PrP/LacZ brain). High resolution section images were obtained using an automated Olympus Bx52 microscope equipped with a high-precision motorized stage running the Neurolucida v6.0 Virtual Slice software (MicroBrightField Inc., Williston, VT, USA).
  
4.&nbsp;&nbsp;&nbsp; Tables listing brain regions with CamKII and / or PrP promoter activity, with specification of corresponding levels in the the image repositories.
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The X-gal staining gives an intense dark-blue labeling at sites with presence of β-galactosidase, which is encoded by the LacZ reporter gene, controlled by the Tet-Off promoters. Presence of labeling thus indicates CamKII or PrP activity, respectively.  
  
Re-use of data from this repository is permitted provided reference is made to the following publication:&nbsp;<br/>Odeh F, Leergaard TB, Boy J, Schmidt T, Riess O, Bjaalie JG (2010), Atlas of transgenic Tet-Off Ca2+/calmodulin-dependent protein kinase II and prion protein promoter activity in the mouse brain. Submitted manuscript.
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All coordinates, annotations and abbreviations used to label and facilitate section identification, are in accordance with the mouse brain atlas by Paxinos and Franklin: The Mouse Brain in Stereotaxic Coordinates, Academic Press, 2001.
  
Re-use of data pertaining to PrP promoter activity should also be referenced to:<br/>Boy J, Leergaard TB, Schmidt T, Odeh F, Bichelmeier U, Nuber S, Holzmann C, Wree A, Prusiner SB, Bujard HB, Riess O, Bjaalie JG (2006), Expression mapping of tetracycline-responsive protein promoter: digital atlasing as a basis for generating cell-specific disease-models. NeuroImage 33: 449-462.
+
== History ==
 +
* 2010-11 TetOff atlas II(PrP/CamKII) released in conjunction with original report by Odeh et al., NeuroImage 54:2603-2611, 2011
 +
* 2009-09 Poster / demonstration of comparative CamKII and PrP promoter activity distributions. 2nd INCF Congress of Neuroinformatics, Plzen, Czeck Republic
 +
* 2006-05 TetOff atlas I(PrP) released. First online digital atlas of the distribution of tetracycline-response protein promoter expression at cellular resolution throughout the mouse brain. Reported in Boy et al., NeuroImage 33: 449-462, 2006
 +
<span style="font-family:georgia,serif">
 +
== Contributing laboratories ==
 +
<div style="clear:both">
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* '''Department of Medical Genetics''', University of Tübingen, Calwerstrasse 7, D-72076 Tübingen, Germany. '''People''': Jana Boy, Thorsten Schmidt, Ulrike Bichelmeier, Silke Nuber, Olaf Riess
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* '''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, development of atlas repository. '''People''': Francis Odeh, Jan O. Kjøde, Ivar A. Moene, Trygve B. Leergaard, Jan G. Bjaalie
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</div>
  
 
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== Funded by ==
 
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* Fritz Thyssen Stiftung
 
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* The Deutsche Heredo-Ataxie-Gesellschaft
 
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* The European Union (6th Framework Programme, EUROSCA)
== News ==
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* The European Union (QLG3-CT-2001-02256)
 
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* The Research Council of Norway
'''2010-09'''&nbsp;&nbsp;TetOff atlas deployed for review in conjunction with submitted original manuscript
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* The neuroinformatics components of this resource were funded by the Human Brain Project through the European Union Seventh Framework Program (FP7/2007-2013) under grant agreement no. 604102 (HBP)<span style="font-family:georgia,serif">
 
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== Contact ==
'''2009-09'''&nbsp; First presentation of CamKII and TetOff atlas for comparing CamKII and PrP promoter activity distributions. Poster/demonstration, 2nd INCF Congress of Neuroinformatics, Plzen, Czeck Republic
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j.g.bjaalie@medisin.uio.no
 
 
'''2006-05'''&nbsp; Release of the PrPpromoter web-application. First online digital atlas of the distribution of tetracycline-response protein promoter expression at cellular resolution throughout the mouse brain. Reported in Boy J et al., NeuroImage 33: 449-462, 2006
 
 
 
'''2004 &nbsp;'''Presentation of the expression patterns of Tet-Off promoter mice in the brain. Poster / demonstration, Eur. J. Hum. Genet. 12, Supp. 1:325, 2004
 
 
 
 
 
 
 
== Tutorial (must be updated) ==
 
 
 
To use the Image repository (Access atlas), first drag the slider below the row of thumbnail images horizontally to select an anterioposterior level of interest (Bregma coordinates are indicated). Click on a thumbnail image to open it in the viewer. The sagittal drawing (right column) indicates the anterioposterior position of the selected image.<br/><br/>The viewer has the following options for navigating the images:
 
 
 
*'''Zoom'''&nbsp;(click the + button in the toolbar or press the shift key to zoom in; click the – button or press the ctrl key to zoom out)
 
*'''Pan&nbsp;'''(click the arrows in the tool bar, or left-click and drag the images freely in the viewer)
 
*'''Reset'''&nbsp;(to reset viewer to default, click the reset button in the toolbar)
 
*'''Annotation&nbsp;'''(currently, no annotations are provided)
 
*'''Scale bar'''&nbsp;(the ZoomScale bar shows a scale bare for the image panels. The bar is transparent and can be moved to make crude measurements in the images)
 
 
 
The right column provides information about the sections, and link to further details about the experiment.&nbsp;<br/><br/>The Bregma value shows the estimated position of the section anterior (positive values) or posterior (negative values) relative to Bregma. The Atlas bregma and Atlas interaural values show the anterioposterior coordinates for the closest diagram in The Rat Brain In Stereotaxic Coordinates (Paxinos and Watson, 2007).&nbsp;<br/><br/>To download images (at viewer resolution) download button in the right column. To print images, either use the print screen functionality of your computer, or use the print function available in the Menu toolbar.
 
 
 
Tables showing distribution of labelled cells:&nbsp;
 
 
 
The distribution tables are opened in separate windows and list brain regions in which promoter activity is seen. It is recommended to use a dual screen setup. For each region containing labeling, the bregma values of the corresponding images are given to facilitate lookup in the image repository. For the tables associated with the CamKII/LacZ and PrP/LacZ repositories, clickable links to the image repository are provided, for the CamKII - PrP comparison atlas this is has not been implemented.
 
 
 
 
 
 
 
FIGURE The Tet-Off atlas is an interactive resource providing access to brain wide histological images showing the distribution of promoter activity in the two most popular transgenic tetracycline-responsive (Tet-Off) promoter mouse lines, the Ca2+/calmodulin-dependent protein kinase II (CamKII) and the prion protein (PrP) mouse lines.
 
 
 
 
 
 
 
FIGURE The atlas has been developed to supplement our earlier atlas of the Tet-Off PrP promoter activity, with that of the Tet-Off CamKII promoter. Thus providing a tool for detailed comparison of the two promoter lines, relevant for generation and analysis of inducible animal models based on these promoter mouse lines.
 
 
 
 
 
 
 
== About the atlas ==
 
 
 
=== Features ===
 
 
 
*Section images (~ 50) showing the distribution of Tet-Off CamKII promoter activity
 
*Section images (~ 50) showing the distribution of Tet-Off PrP promoter activity (previously reported in Boy et al., NeuroImage 33: 449-462, 2006
 
*Dual image viewer tool for side-by-side comparison of spatially corresponding section images showing Tet-Off CamKII and PrP promoter activity
 
*Table listing brain regions with Tet-Off CamKII and / or PrP promoter activity, with region-specific links to the image repository
 
 
 
 
 
 
 
=== Methods ===
 
 
 
The atlas is based on data from two double transgenic mice, a 40 week old female CamKII/LacZ mouse and a 5 week old female PrP/LacZ mouse (Boy et al., NeuroImage 33: 449-462, 2006). For methodological details concerning gene construct, preparation of histological material, image acquisition, and atlas construction, see Boy et al., NeuroImage 33: 449-462, 2006; Odeh et al., submitted manuscript, 2010). In brief, double-transgenic mice were generated by cross-breeding a CamKII or PrP promoter mouse line with a LacZ responder mouse line. Animals were anaesthetized and transcardially perfused with 4% paraformaledhyde before brains were removed and stained using X-Gal (to label the LacZ product&nbsp;?-galactosidase) and counterstained with Cresyl violet (sections from the CamKII/LacZ brain) or Neutral red (sections from the PrP/LacZ brain). High resolution section images were obtained using an automated Olympus Bx52 microscope equipped with a high-precision motorized stage running the Neurolucida v6.0 Virtual Slice software (MicroBrightField Inc., Williston, VT, USA).
 
 
 
The X-gal staining gives an intense dark-blue labeling at sites with presence of&nbsp;?-galactosidase, which is encoded by the LacZ reporter gene, controlled by the Tet-Off promoters. Presence of labeling thus indicates CamKII or PrP activity, respectively. &nbsp;
 
 
 
 
 
 
 
FIGURE&nbsp;Example of an X-gal stained section from a CamKII/LacZ mouse (dark blue cells indicated by arrowheads), counterstained with Cresyl violet (brightly stained violet cells)
 
 
 
 
 
 
 
FIGURE&nbsp;Example of an X-gal stained section from a PrP/LacZ mouse (dark blue cells indicated by arrowheads), counterstained with Neutral red (brightly stained red cells)
 
 
 
 
 
References ===
 
 
 
''PrP atlas application:''<br/>Boy J, Leergaard TB, Schmidt T, Odeh F, Bichelmeier U, Nuber S, Holzmann C, Wree A, Prusiner SB, Bujard HB, Riess O, Bjaalie JG (2006), Expression mapping of tetracycline-responsive protein promoter: digital atlasing as a basis for generating cell-specific disease-models. NeuroImage 33: 449-462.
 
 
 
''Tet-Off atlas and CamKII atlas application:''<br/>Odeh F, Leergaard TB, Boy J, Schmidt T, Riess O, Bjaalie JG (2010), Atlas of transgenic Tet-Off Ca2+/calmodulin-dependent protein kinase II and prion protein promoter activity in the mouse brain. Submitted manuscript.&nbsp;
 
 
 
''Database architecture:''<br/>Moene IA, Subramaniam S, Darine D, Leergaard TB, Bjaalie JG (2007). Towards a workbench for rodent brain image data: systems architecture and design. Neuroinformatics; 5: 35-58&nbsp;
 
 
 
Bjaalie JG, Leergaard TB, Lillehaug S, Odeh F, Moene IA, Kjøde JO, Darine D (2005) Database and tools for analysis of topographic organization and map transformations in major projection systems of the brain. Neuroscience 2005; 136:681-696
 

Revision as of 12:16, 25 January 2018

About

The tetracycline-responsive (TetR) gene system is a binary transgenic system in which transgene expression may be activated or silenced by administration of tetracycline, or its derivates (Gossen and Bujard, PNAS 89:5547-5551, 1992; Gossen et al., Science 286:1766-1769, 1995). Such animal models are generated by crossing two different mouse lines, one with a gene modulator (promoter mouse line) and one with the actual disease-causing gene (responder mouse line). In the double transgenic offspring both the regional distribution and intensity of target gene expression depend on the activity of the chosen promoter controlling the expression of a gene activator, which in turn is accessible for experimental modulation. Detailed knowledge about where in the brain gene expression is regulated is essential for the use and interpretation of such models.

The TetOff atlas 2 (PrP/CamKII) is an interactive resource providing access to a collection of microscopic brain sections displaying the spatial distribution patterns of promoter activity of two popular transgenic tetracycline-responsive (Tet-Off) promoter mouse lines, the prion protein (PrP) mouse line (previously reported in Boy et al., Neuroimage 33:449-462, 2006) and the Ca2+/calmodulin-dependent protein kinase II (CamKII).  

Access image repository

The virtual microscopy viewer allows interactive zooming and panning. Original images are available for download via separate link. Each section has been annotated with names of key landmarks, regions, nuclei, and areas.

Re-use of data from this repository is allowed provided that reference is given to the following publications: 

  • CamKII: Odeh F, Leergaard TB, Boy J, Schmidt T, Riess O, Bjaalie JG. Atlas of transgenic Tet-Off Ca2+/calmodulin-dependent protein kinase II and prion protein promoter activity in the mouse brain. NeuroImage 54:2603-2611, 2011 
  • PrP: Boy J, Leergaard TB, Schmidt T, Odeh F, Bichelmeier U, Nuber S, Holzmann C, Wree A, Prusiner SB, Bujard HB, Riess O, Bjaalie JG. Expression mapping of tetracycline-responsive prion protein promoter: digital atlasing for generating cell-specific disease-models. NeuroImage 33:449-462, 2006 
Animal # Genotype Image repository Download original
388.12 Prnp-tTA:βGal Filmstrip viewer Tiffs
317.8 CamKII-tTA:βGal Filmstrip viewer Tiffs

Experimental procedures in brief

The atlas is based on data from two double transgenic mice, a 40 week old female CamKII/LacZ mouse and a 5 week old female PrP/LacZ mouse (Boy et al., NeuroImage 33: 449-462, 2006). For methodological details concerning gene construct, preparation of histological material, image acquisition, and atlas construction, see Boy et al. (NeuroImage, 33:449-462, 2006), and Odeh et al. (NeuroImage 54:2603-2611, 2011).

In brief, double-transgenic mice were generated by cross-breeding a CamKII or PrP promoter mouse line with a LacZ responder mouse line. Animals were anaesthetized and transcardially perfused with 4% paraformaledhyde before brains were removed and stained using X-Gal (to label the LacZ product β-galactosidase) and counterstained with Cresyl violet (sections from the CamKII/LacZ brain) or Neutral red (sections from the PrP/LacZ brain). High resolution section images were obtained using an automated Olympus Bx52 microscope equipped with a high-precision motorized stage running the Neurolucida v6.0 Virtual Slice software (MicroBrightField Inc., Williston, VT, USA).

The X-gal staining gives an intense dark-blue labeling at sites with presence of β-galactosidase, which is encoded by the LacZ reporter gene, controlled by the Tet-Off promoters. Presence of labeling thus indicates CamKII or PrP activity, respectively.  

All coordinates, annotations and abbreviations used to label and facilitate section identification, are in accordance with the mouse brain atlas by Paxinos and Franklin: The Mouse Brain in Stereotaxic Coordinates, Academic Press, 2001.

History

  • 2010-11 TetOff atlas II(PrP/CamKII) released in conjunction with original report by Odeh et al., NeuroImage 54:2603-2611, 2011
  • 2009-09 Poster / demonstration of comparative CamKII and PrP promoter activity distributions. 2nd INCF Congress of Neuroinformatics, Plzen, Czeck Republic
  • 2006-05 TetOff atlas I(PrP) released. First online digital atlas of the distribution of tetracycline-response protein promoter expression at cellular resolution throughout the mouse brain. Reported in Boy et al., NeuroImage 33: 449-462, 2006

Contributing laboratories

  • Department of Medical Genetics, University of Tübingen, Calwerstrasse 7, D-72076 Tübingen, Germany. People: Jana Boy, Thorsten Schmidt, Ulrike Bichelmeier, Silke Nuber, Olaf Riess
  • 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, development of atlas repository. People: Francis Odeh, Jan O. Kjøde, Ivar A. Moene, Trygve B. Leergaard, Jan G. Bjaalie

Funded by

  • Fritz Thyssen Stiftung
  • The Deutsche Heredo-Ataxie-Gesellschaft
  • The European Union (6th Framework Programme, EUROSCA)
  • The European Union (QLG3-CT-2001-02256)
  • The Research Council of Norway
  • The neuroinformatics components of this resource were 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