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Anatomical Science International

, Volume 93, Issue 4, pp 414–421 | Cite as

From the primitive streak to the somitic mesoderm: labeling the early stages of chick embryos using EGFP transfection

  • Haiming Fan
  • Nobuyuki Sakamoto
  • Hirohiko Aoyama
Original Article
  • 148 Downloads

Abstract

Mesoderm is derived from the primitive streak. The rostral region of the primitive streak forms the somitic mesoderm. We have previously shown the developmental origin of each level of the somitic mesoderm using DiI fluorescence labeling of the primitive streak. We found that the more caudal segments were derived from the primitive streak during the later developmental stages. DiI labeled several pairs of somites and showed the distinct rostral boundary; however, the fluorescence gradually disappeared in the caudal region. This finding can be explained in two ways: the primitive streak at a specific developmental stage is primordial of only a certain number of pairs of somites, or the DiI fluorescent dye was gradually diluted within the primitive streak by cell division. Here, we traced the development of the primitive streak cells using enhanced green fluorescent protein (EGFP) transfection. We confirmed that, the later the EGFP transfection stage, the more caudal the somites labeled. Different from DiI labeling, EGFP transfection performed at any developmental stage labeled the entire somitic mesoderm from the anterior boundary to the tail bud in 4.5-day-old embryos. Furthermore, the secondary neural tube was also labeled, suggesting that not only the somite precursor cells but also the axial stem cells were labeled.

Keywords

Chick Developmental fate In ovo electroporation Primitive streak Somite 

Notes

Acknowledgements

We thank Dr. Yoshiko Takahashi of Kyoto University for kindly providing pT2 K-CAGGS-EGFP and pCAGGS-T2TP. A part of this study was supported by Grants-in-Aid for Scientific Research from JSPS (20590171, 23590219, 26460254).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

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Copyright information

© Japanese Association of Anatomists 2018

Authors and Affiliations

  • Haiming Fan
    • 1
  • Nobuyuki Sakamoto
    • 1
  • Hirohiko Aoyama
    • 1
  1. 1.Department of Anatomy and Developmental Biology, Graduate School of Biomedical SciencesHiroshima UniversityHiroshimaJapan

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