Difference between revisions of "Brain Atlas of tTA driver lines"

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Revision as of 23:27, 24 January 2018

About

The Brain Atlas of tTA driver lines is an interactive resource providing access to a comprehensive collection of microscopic images showing brain-wide reporter expression for five commonly used tTA driver lines mapped to a common reference atlas space. The tetracycline dependent regulatory system is the most frequently used system for conditional regulation of gene expression, in which temporally-regulated transgene expression is achieved by exogenous control of transcription by administration of tetracycline or its chemical derivatives (Gossen and Bujard, PNAS, 89:5547-51, 1992; Lewandoski, Nat Rev Genet 2, 743-55, 2001). Tetracycline-responsive transcription depends on a bipartite system in which the promoter controlling a transgene of interest is activated exclusively by an artificial fusion protein known as the tetracycline-controlled transactivator (tTA). The promoter used to restrict tTA expression therefore dictates the spatial distribution of the transgenic protein of interest.

Image repository

The image repository contains ~450 microscopic images showing data from 12 bigenic tTA/tetO-lacZ constructs representing 5 tA driver lines, and one control animal. The virtual microscopy viewer (LocaliZoom) allows interactive zooming (mouse wheel) and panning (right-click and drag), adjustment of atlas overlay transparency (slider), and shows atlas coordinates at the mouse pointer in real-time. Point coordinates can be annotated using space button, and may exported as X,Y,Z coordinate lists. A link is also provided to a geometric mesh viewer tool (MeshView), which provides 3D rendering of anatomical regions delineated in the Allen mouse brain atlas (v2) and allows viewing annotated points (paste into Coordinates window). In the MeshView viewer tool (www.nitrc.org), atlas regions can be rendered as solid, transparent, or invisible objects, and cut in user defined planes.

Original images are available for download via separate link. The file names encode the animal number (first four digits), genotype, staining (X-gal), serial number (_s followed by three digits), and pixel dimensions (extension _1.4 or _0.22, indicating µm/pixel).

The amount of X-gal-labeled cells across different brain regions in all animals was semi-quantitatively scored using a grading system from 0-4, with grade 0 representing absence of labeled cells, grade 1 low density (few cells, possible to count), grade 2 (medium density, cells can be individually discerned but not readily counted), grade 3 (many, largely overlapping cells), and grade 4 (very high density, individual cells cannot be discerned).

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

  • Pcp2-tTA, Pitx3-tTA, and online repository with and images anchored to atlas space: Lillehaug, S. et al. (submitted manuscript). Repository of tetracycline-responsive promoter expression distributions mapped in mouse brain atlas space.
  • Nop-tTA-lacZ: Yetman, M. et al. (2016) Transgene expression in the Nop-tTA driver line is not inherently restricted to the entorhinal cortex. Brain Struct Funct 221, 2231-2249, doi:10.1007/s00429-015-1040-9
  • CaMKII-tTA-lacZ: Odeh, F. et al. (2011). Atlas of transgenic Tet-Off Ca2+/calmodulin-dependent protein kinase II and prion protein promoter activity in the mouse brain. NeuroImage 54, 2603-2611, doi:10.1016/j.neuroimage.2010.11.032
  • PrP-tTA-lacZ: Boy, J. et al. (2006). Expression mapping of tetracycline-responsive prion protein promoter: digital atlasing for generating cell-specific disease models. NeuroImage 33, 449-462, doi:10.1016/j.neuroimage.2006.05.055
Driver line / Reporter line Animal # Orientation Image repository Download original
Pcp2 Pcp2-tTA/tetO-lacZ-nls-GFP 1261 coronal LocaliZoom 41 tiff images
Pcp2-tTA/tetO-lacZ-nls-GFP 3292 horizontal LocaliZoom 23 tiff images
Pcp2-tTA/tetO-lacZ-nls-GFP 4340 coronal LocaliZoom 33 tiff images
Pitx3 Pitx3-tTA/tetO-lacZ-nls-GFP 3435 coronal LocaliZoom 42 tiff images
Pitx3-tTA/tetO-lacZ-nls-GFP 5154 horizontal LocaliZoom 16 tiff images
Pitx3-tTA/tetO-lacZ-nls-GFP 6513 horizontal LocaliZoom 15 tiff images
Pitx3-tTA/tetO-lacZ-nls-GFP 6517 coronal LocaliZoom 32 tiff images
Nop Nop-tTA/tetO-lacZ-nls-GFP 1952 coronal LocaliZoom 45 tiff images
Nop-tTA/tetO-lacZ-nls-GFP 2849 coronal LocaliZoom 34 tiff images
Nop-tTA/tetO-lacZ-nls-GFP 2877 horizontal LocaliZoom 23 tiff images
CaMKII CaMKII-tTA/tetO-lacZ 317.8 coronal LocaliZoom 47 tiff images
PrP Prnp-tTA/tetO-lacZ 388.12 coronal LocaliZoom 57 tiff images
Control tetO-lacZ-nls-GFP 1950 coronal Filmstrip 49 tiff images

Experimental procedures in brief

The atlas is based on data from 14 double-transgenic mice. For details concerning gene construct, preparation of histological materials, image acquisition, and atlas construction, see publications listed above (Boy et al., 2006; Odeh et al., 2011; Yetman et al., 2016; Lillehaug et al., 2018). In brief, mice were transcardially perfused with 4% paraformaldehyde, brains removed and sectioned coronally or horizontally using a cryo-microtome. Sections were stained using X-Gal to label the LacZ product β-galactosidase (yielding an intense dark-blue labeling) and counterstained to visualize the gross cytoarchitecture. Section images acquired through a 20× objective either an Olympus Bx52 motorized microscope running the Neurolucida v6.0 Virtual Slice software (MicroBrightField Inc., Williston, VT, USA) or a Zeiss Axioscan Z1 slide scanner running Zen software (Carl Zeiss MicroImaging, Jena, Germany). For each image a spatially matching, custom made slice through the Allen institute mouse brain atlas (v2; http://mouse.brain-map.org) is provided as an overlay image. For further methodological details, cf. original papers (listed above).

History

  • Brain atlas of tTA driver lines: collection of image data from 5 tTA driver lines (Prp, CaMKII, Nop, Pitx3, Pcp2) spatially defined in the Allen mouse brain reference atlas, described in Lillehaug et al., submitted manuscript, 2018
  • tTA / TetOff atlas: Nop, reported in Yetman et al. Brain Struct Funct 221:2231-2249, 2016
  • tTA / TetOff atlas: PrP/CaMKII, released in conjunction with original report by Odeh et al., NeuroImage 54:2603-2611, 2011
  • tTA / TetOff atlas: 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

Departments of Neuroscience (https://www.bcm.edu/centers/huffington-center-on-aging/), Huffington Center on Aging, Baylor College of Medicine, Houston, TX, USA and Departments of Neurology and Neurosurgery, Huffington Center on Aging, Baylor College of Medicine, Houston, TX, USA. People: Michael J. Yetman, Joanna L. Jankowsky.

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: Sveinung Lillehaug, Maja A. Puchades, Martyna M. Checinska, Francis Odeh, Jan O. Kjøde, Ivar A. Moene, Gergely Csucs, Dmitri Darine, Trygve B. Leergaard & Jan G. Bjaalie

Department of Medical Genetics (http://www.medgen-tuebingen.de/start.html), University of Tübingen, Calwerstrasse 7, D-72076 Tübingen, Germany. People: Jana (Boy) Schmidt, Thorsten Schmidt, Ulrike Bichelmeier, Silke Nuber, Olaf Riess.

Funded by

  • European Union`s Horizon 2020 Research and Innovation Programme under Grant Agreement No. 720270 (HBP SGA1, to Jan G. Bjaalie, Trygve B. Leergaard)
  • National Institutes of Health (NIH) Office of the Director (DP2 OD001734 to Joanna L. Jankowsky)
  • National Institute of Aging (T32 AG000183 to Michael J. Yetman)
  • The Research Council of Norway (214842 and the Norwegian Large Scale Infrastructure for Brain Research; NORBRAIN to Jan G. Bjaalie)
  • The European Union (QLG3-CT-2001-02256 to Jan G. Bjaalie)
  • Fritz Thyssen Stiftung (to Olaf Riess)
  • The Deutsche Heredo-Ataxie-Gesellschaft (to Olaf Riess and Thorsten Schmidt)
  • The European Union (6th Framework Programme, EUROSCA to Olaf Riess)

Contacts

j.g.bjaalie@medisin.uio.no

t.b.leergaard@medisin.uio.no