Transgenesis of the Wolffian duct visualizes dynamic behavior of cells undergoing tubulogenesis in vivo

Yuji Atsuta, Ryosuke Tadokoro, Daisuke Saito, Yoshiko Takahashi

研究成果: Article査読

13 被引用数 (Scopus)

抄録

Deciphering how the tubulogenesis is regulated is an essential but unsolved issue in developmental biology. Here, using Wolffian duct (WD) formation in chicken embryos, we have developed a novel method that enables gene manipulation during tubulogenesis in vivo. Exploiting that WD arises from a defined site located anteriorly in the embryo (pronephric region), we targeted this region with the enhanced green fluorescent protein (EGFP) gene by the in ovo electroporation technique. EGFP-positive signals were detected in a wide area of elongating WD, where transgenic cells formed an epithelial component in a mosaic manner. Time-lapse live imaging analyses further revealed dynamic behavior of cells during WD elongation: some cells possessed numerous filopodia, and others exhibited cellular tails that repeated elongation and retraction. The retraction of the tail was precisely regulated by Rho activity via actin dynamics. When electroporated with the C3 gene, encoding Rho inhibitor, WD cells failed to contract their tails, resulting in an aberrantly elongated process. We further combined with the Tol2 transposon-mediated gene transfer technique, and could trace EGFP-positive cells at later stages in the ureteric bud sprouting from WD. This is the first demonstration that exogenous gene(s) can directly be introduced into elongating tubular structures in living amniote embryos. This method has opened a way to investigate how a complex tubulogenesis proceeds in higher vertebrates. Development, Growth & Differentiation

本文言語English
ページ(範囲)579-590
ページ数12
ジャーナルDevelopment Growth and Differentiation
55
4
DOI
出版ステータスPublished - 2013 5

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

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