Hallgrimsson1 project protocol

Facial shape and allometry in a Diversity Outbred (DO) mouse population   (2020)

Katz DC, Hallgrimsson B
With: Aponte JD, Liu W, Green RM, Mayeux JM, Pollard KM, Pomp D, Munger SC, Murray SA, Roseman CC, Percival CJ, Cheverud J, Marcucio RS




Project protocol - Contents

Workflow and sampling

Step Description Equipment Data collected
1 Mice sacrificed and weighed Balance Body weight
2 Skulls prepared for image analysis Image detection and analysis software Facial shape 3D landmark coordinate data

Procedures

Procedure 1: Sample preparation

Equipment, software, and supplies

  • Balance

Steps

  1. DO mice were raised in several laboratories. The DO sample consists of approximately 1147 male and female adult mice from seven DO generations, which are 9, 10, 15, 19, 21, 23 and 27.
  2. Mice are sacrificed and weighed, after which skulls are isolated and prepared for imaging.

Procedure 2: Images and landmark data acquisition

Equipment, software, and supplies

  • Scanco in-vivo microCT (micro-computed tomography) scanner (Scanco Medical, vivaCT40, Bruttisellen, Switzerland)
  • Analyze 3D, image visualization software (AnalyzeDirect Inc., Overland Park, KS, USA)

Environmental Conditions

Holding Area and/or Sample Collection Area

Facility: 3D Morphometrics Centre at the University of Calgary Foothills Campus, Calgary, Alberta, Canada

Steps

  1. microCT images of mouse skulls are obtained using a Scanco vivaCT40 scanner at 0.035 mm voxel dimensions at 55kV and 72-145 μA.
  2. An operator collected 54, 3D landmark coordinates from minimum threshold-defined bone surface, from each skull with ANALYZE 3D image visualization software. A 22-landmark subset of the full configuration were measured in three dimensions, giving 66 coordinates.

References

Primary References

Katz DC, Aponte JD, Liu W, Green RM, Mayeux JM, Pollard KM, Pomp D, Munger SC, Murray SA, Roseman CC, Percival CJ, Cheverud J, Marcucio RS, Hallgrímsson B. Facial shape and allometry quantitative trait locus intervals in the Diversity Outbred mouse are enriched for known skeletal and facial development genes. PLoS One. 2020;15(6):e0233377. doi: 10.1371/journal.pone.0233377. Epub 2020 Jun 5.   PubMed 32502155     FullText