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

Revision as of 14:33, 6 November 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. This image repository holds serial microscopic images showing brain-wide reporter expression for five commonly used tTA driver lines:

• L7/Purkinje cell protein 2 (Pcp2)-tTA/tetO-lacZ-nls-GFP mice (Pcp2-tTA)
• Pituitary homeobox 3(Pitx3)-tTA/tetO-lacZ-nls-GFP mice (Pitx3-tTA)
• PrP-tTA/tetO-lacZ (Prp-tTA)
• CaMKII-tTA/tetO-lacZ (CaMKII-tTA)
• Nop-tTA/tetO-lacZ-nls-GFP (Nop-tTA) mice 

All images have been spatially registered to the Allen Mouse Common Coordinate Framework (CCF) version 2 (Oh, S.W., et al., Nature, 508:7495 207, 2014) using the QuickNII software tool. This allows direct comparison of anatomical distribution across lines (see Overview Table, below).

Image repository

The image repository contains ~450 microscopic images showing data from 12 bigenic tTA/tetO-lacZ constructs representing 5 tA driver lines.

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 CCF v.2 and allows viewing of 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).

Overview table

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).

License and data citation

Creative Commons Attribution-ShareAlike 4.0 International

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

Experimental procedures 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 Mouse CCF v.2  is provided as an overlay image. For further methodological details, cf. original papers (listed above).

For details concerning gene construct, preparation of histological materials, image acquisition, and atlas construction, see publications listed above (Licence and data citation).

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

t.b.leergaard@medisin.uio.no

j.g.bjaalie@medisin.uio.no