TTA / TetOff Atlas (PrP)

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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 TetOff atlas 1 (PrPpromoter) is an interactive resource providing access to a collection of microscopic brain section from a double transgenic mouse line (Prnp-tTA:βGal), obtained by crossbreeding a tetracycline-responsive prion protein promoter mouse line with a responder mouse line transgenic for a reporter gene construct containing the LacZ reporter gene (encoding the enzyme β-galactosidase).

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 publication: 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 prp-lacZ Filmstrip viewer Tiffs

Experimental procedures in brief

Double transgenic mice were produced by crossbreeding a Prnp-tTA mouse line containing the hamster PrP promoter gene (Tremblay et al., PNAS 95:12580-12585,1998) with a responder mouse line transgenic for a bidirectional reporter gene construct containing both the Luciferase and LacZ reporter gene (Baron et al., Nucleic Acids Res. 23:3605-3606, 1995), which encodes the enzyme β-galactosidase. In the resulting double transgenic mice (Prnp-tTA:βGal) inducible LacZgene expression is under control of the PrP gene (Prnp) promoter. The PrP promoter was obtained from hamster, theLuciferase gene (which was not employed in this study) from Photinus pyralis, and the LacZ gene from E. coli. The LacZgene product (β-galactosidase) was identified using X-gal (5-Bromo-4-chloro-3-indolyl β-D-galactopyranoside) as a substrate.

Anaesthetized mice were transcardially perfused with 4% paraformaldehyde. Dissected whole brains were sectioned at 25 µm on a cryostat. Every eighth consecutive section (200 µm spacing) was collected for processing as free-floating sections with X-gal. Stained sections were mounted on glass slides, counterstained with Neutral Red, and coverslipped.

High-resolution mosaic section images were obtained through an automated Olympus Bx52 microscope, equipped with a high-precision motorized stage (LEP MAC5000, LUDL Electronic Products Ltd., Hawthorne, NY, USA), an Optronics MicroFire digital camera (Optronics Picture This, Goleta, CA USA), and Neurolucida v6.0 Virtual Slice software (MicroBrightField Inc., Williston, VT, USA).

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.

For further details, see 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
  • Department of Medical Genetics, University of Rostock, Rembrandtstrasse 16/17, D-18057 Rostock, Germany. People: Carsten Holzmann
  •  Department of Anatomy, University of Rostock, Gertrudenstrasse 9, D - 18057 Rostock, Germany. People: Andreas Wree
  •  Institute for Neurodegenerative Diseases, University of California, San Franscisco, Box 0518 San Francisco, CA 94143-0518, USA. People: Stanley Prusiner
  •  ZMBH (Zentrum für Molekulare Biologie Heidelberg), University of Heidelberg, Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany. People: Hermann Bujard
  •  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: 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 (Marie Curie EST programme, HPMT-CT-2001-00406-03)
  • The European Union (QLG3-CT-2001-02256)
  • The Research Council of Norway
  • The neuroinformatics components of this resource was 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