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

Yuji Atsuta, Ryosuke Tadokoro, Daisuke Saito, Yoshiko Takahashi

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

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

Original languageEnglish
Pages (from-to)579-590
Number of pages12
JournalDevelopment Growth and Differentiation
Volume55
Issue number4
DOIs
Publication statusPublished - May 1 2013
Externally publishedYes

Fingerprint

Wolffian Ducts
Gene Transfer Techniques
Tail
Embryonic Structures
Genes
Time-Lapse Imaging
Developmental Biology
Pseudopodia
Electroporation
Growth and Development
Vertebrates
Actins
Chickens
Epithelial Cells

All Science Journal Classification (ASJC) codes

  • Developmental Biology
  • Cell Biology

Cite this

Transgenesis of the Wolffian duct visualizes dynamic behavior of cells undergoing tubulogenesis in vivo. / Atsuta, Yuji; Tadokoro, Ryosuke; Saito, Daisuke; Takahashi, Yoshiko.

In: Development Growth and Differentiation, Vol. 55, No. 4, 01.05.2013, p. 579-590.

Research output: Contribution to journalArticle

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