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

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Revision as of 10:05, 7 November 2018

About

The transgene expression in tetracycline-transactivator 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:

  • Ca2+/calmodulin-dependent protein kinase II (CaMKII)-tTA/tetO-lacZ (CaMKII-tTA)
  • Neuropsin (Nop)-tTA/tetO-lacZ-nls-GFP (Nop-tTA) mice 
  • 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) 
  • Prion promoter (Prp)-tTA/tetO-lacZ (Prp-tTA)

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 images are hosted by the EU Human Brain Project (www.humanbrainproject.eu). Each data set has been assigned a Data Cite Digital Object Identifiyer (DOI).

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.

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).

For each microscopic image, an atlas map is provided as a 1024 pixel-wide PNG image that represents the same physical space (i.e. with matching corners) as the higher resolution microscopic images. The atlas maps are named to match their corresponding TIFF images with an additional extension (_segmentation).

Driver line / Reporter line Animal # Gender Age (weeks) Orientation Image repository Download original images View atlas maps Download atlas maps Data set DOI
CaMKII CaMKII-tTA/tetO-lacZ 317.8 female 41 coronal LocaliZoom 47 tiff images 47 png images CaMKII 317.8 10.25493/W97C-01R
Nop Nop-tTA/tetO-lacZ-nls-GFP 1952 male 38,5 coronal LocaliZoom 45 tiff images 45 png images Nop 1952 10.25493/5H13-1Q0
Nop-tTA/tetO-lacZ-nls-GFP 2877 male 36 horizontal LocaliZoom 23 tiff images 23 png images Nop 2877 10.25493/AYBB-BXV
Nop-tTA/tetO-lacZ-nls-GFP 2849 female 31 coronal LocaliZoom 34 tiff images 32 png images* Nop 2849 10.25493/WB6K-V72
Pcp2 Pcp2-tTA/tetO-lacZ-nls-GFP 1261 male 42,5 coronal LocaliZoom 41 tiff images 36 png images* Pcp2 1261 10.25493/A2EG-VPR
Pcp2-tTA/tetO-lacZ-nls-GFP 3292 female 37 horizontal LocaliZoom 23 tiff images 23 png images Pcp2 3292 10.25493/D1PR-55A
Pcp2-tTA/tetO-lacZ-nls-GFP 4340 male 20 coronal LocaliZoom 33 tiff images 29 png images* Pcp2 4340 10.25493/YCGE-ADC
Pitx3 Pitx3-tTA/tetO-lacZ-nls-GFP 3435 male 15 coronal LocaliZoom 42 tiff images 42 png images Pitx3 3435 10.25493/FMGJ-4N3
Pitx3-tTA/tetO-lacZ-nls-GFP 5154 female 12,5 horizontal LocaliZoom 16 tiff images 16 png images Pitx3 5154 10.25493/2382-AYM
Pitx3-tTA/tetO-lacZ-nls-GFP 6513 male 9 horizontal LocaliZoom 15 tiff images 15 png images Pitx3 6513 10.25493/1AH7-T1A
Pitx3-tTA/tetO-lacZ-nls-GFP 6517 female 9 coronal LocaliZoom 32 tiff images 32 png images Pitx3 6517 10.25493/DTB3-0KP
PrP Prnp-tTA/tetO-lacZ 388.12 female 5 coronal LocaliZoom 57 tiff images 57 png images PrP 388.12 10.25493/T9P1-WH4

*The discrepancy in corresponding image and atlas map has been accounted for.

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).

Comparative overview of brain-wide tetracycline transactivator expression across driver lines Access PDF (external link) Access CSV (external link) Data set DOI: 10.25493/ARKS-R7H

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