|ZFIN ID: ZDB-PUB-170510-22|
Utility of quantitative micro-computed tomographic analysis in zebrafish to define gene function during skeletogenesis
Charles, J.F., Sury, M., Tsang, K., Urso, K., Henke, K., Huang, Y., Russell, R., Duryea, J., Harris, M.P.
|Source:||Bone 101: 162-171 (Journal)|
|Registered Authors:||Harris, Matthew, Henke, Katrin|
|Keywords:||Bone, Bone matrix specific contrast, Development, Microcomputed tomography, Phenotypic variation, Zebrafish|
|PubMed:||28476577 Full text @ Bone|
Charles, J.F., Sury, M., Tsang, K., Urso, K., Henke, K., Huang, Y., Russell, R., Duryea, J., Harris, M.P. (2017) Utility of quantitative micro-computed tomographic analysis in zebrafish to define gene function during skeletogenesis. Bone. 101:162-171.
ABSTRACTThe zebrafish is a powerful experimental model to investigate the genetic and morphologic basis of vertebrate development. Analysis of skeletogenesis in this fish is challenging as a result of the small size of the developing and adult zebrafish. Many of the bones of small fishes such as the zebrafish and medaka are quite thin, precluding many standard assays of bone quality and morphometrics commonly used on bones of larger animals. Microcomputed tomography (microCT) is a common imaging technique used for detailed analysis of the skeleton of the zebrafish and determination of mutant phenotypes. However, the utility of this modality for analysis of the zebrafish skeleton, and the effect of inherent variation among individual zebrafish, including variables such as sex, age and strain, is not well understood. Given the increased use and accessibility of microCT, we set out to define the sensitivity of microCT methods in developing and adult zebrafish. We assessed skeletal shape and density measures in the developing vertebrae and parasphenoid of the skull base. We found most skeletal variables are tightly correlated to standard length, but that at later growth stages (>3months) there are age dependent effects on some skeletal measures. Further we find modest strain but not sex differences in skeletal measures. These data suggest that the appropriate control for assessing mutant phenotypes should be age and strain matched, ideally a wild-type sibling. By analyzing two mutants exhibiting skeletal dysplasia, we show that microCT imaging can be a sensitive method to quantify distinct skeletal parameters of adults. Finally, as developing zebrafish skeletons remain difficult to resolve by radiographic means, we define a contrast agent specific for bone that enhances resolution at early stages, permitting detailed morphometric analysis of the forming skeleton. This increased capability for detection extends the use of this imaging modality to leverage the zebrafish model to understand the development causes of skeletal dysplasias.