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Open Access Methodology article

In ovo serial skeletal muscle diffusion tractography of the developing chick embryo using DTI: feasibility and correlation with histology

Zien Zhou1, Zachary DelProposto2, Lianming Wu13, Jianrong Xu1*, Jia Hua1, Yan Zhou1, Yongquan Ye3, Zishu Zhang3, Jiani Hu3 and E Mark Haacke3

Author Affiliations

1 Department of Radiology, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China

2 Department of Radiology, Henry Ford Hospital, Detroit, MI, USA

3 Department of Radiology, Wayne State University, Detroit, MI, USA

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BMC Developmental Biology 2012, 12:38  doi:10.1186/1471-213X-12-38

Published: 26 December 2012

Abstract

Background

Magnetic resonance imaging is a noninvasive method of evaluating embryonic development. Diffusion tensor imaging (DTI), based on the directional diffusivity of water molecules, is an established method of evaluating tissue structure. Yet embryonic motion degrades the in vivo acquisition of long-duration DTI. We used a dual-cooling technique to avoid motion artifact and aimed to investigate whether DTI can be used to monitor chick embryonic skeletal muscle development in ovo, and to investigate the correlation between quantitative DTI parameters fractional anisotropy (FA) and fiber length and quantitative histologic parameters fiber area percentage (FiberArea%) and limb length.

Results

From 84 normally developing chick embryos, 5 were randomly chosen each day from incubation days 5 to 18 and scanned using 3.0 Tesla magnetic resonance imaging. A dual-cooling technique is used before and during imaging. Eggs were cracked for making histological specimen after imaging. 3 eggs were serially imaged from days 5 to 18. We show that skeletal muscle fibers can be tracked in hind limb in DTI beginning with incubation day 8. Our data shows a good positive correlation between quantitative DTI and histologic parameters (FA vs FiberArea%: r= 0.943, p<0.0001; Fiber_length vs Limb_length: r=0.974, p<0.0001). The result of tracked fibers in DTI during incubation corresponds to the development of chick embryonic skeletal muscle as reported in the literature.

Conclusion

Diffusion tensor imaging can provide a noninvasive means of evaluating skeletal muscle development in ovo.

Keywords:
Magnetic resonance imaging; Diffusion tensor imaging; Skeletal muscle; Chick embryo