Related Experiment Video
Updated: Dec 6, 2025
Optimized Ex-ovo Culturing of Chick Embryos to Advanced Stages of Development
Published on: January 24, 2015
Teratogenesis in the chick embryo following post-gastrulation exposure to Y-27632 -effect of Y-27632 on embryonic
Johannes W Duess1, Jan-Hendrik Gosemann2, Prem Puri3
1Department of Pediatric Surgery, University of Leipzig, Leipzig, Germany; National Children's Research Centre, Our Lady's Children's Hospital, Crumlin, Dublin 12, Ireland; School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4, Ireland.
Abstract:
The pyridine derivative Y-27632 inhibits Rho-associated coiled-coil-containing protein kinase (ROCK) signaling, which is involved in numerous developmental processes during embryogenesis, primarily by controlling actin-cytoskeleton assembly and cell contractility. Somite formation requires rearrangement of the cytoskeleton and assists in major morphological mechanisms, including ventral body wall formation. Administration of Y-27632 impairs cytoskeletal arrangements in post-gastrulation chick embryos leading to ventral body wall defects (VBWD) at later stages of development. The aim of this study was to investigate the effect of Y-27632 on somite development in post-gastrulation chick embryos during early embryogenesis. After 60 h incubation, embryos in shell-less culture were treated with Y-27632 or vehicle for controls. Following administration, abnormality rates were assessed. In treatment groups, embryos showed a kinked longitudinal body axis. Western blot confirmed impaired ROCK downstream signaling by decreased expression of phosphorylated cofilin-2. Histology, Lysotracker studies and RT-PCR demonstrated increased cell death in somites, the neural tube and the ectoderm. RT-PCR and Western blot of factors known to be involved during somitogenesis revealed reduced expression in the treatment group compared to controls. We hypothesize that administration of Y-27632 disrupts somite development causing axial kinking and embryo malformation, which may lead to VBWD.
Insights
Y-27632 disrupts embryonic development by inhibiting Rho-associated coiled-coil-containing protein kinase (ROCK) signaling, leading to somite abnormalities and axial kinking in chick embryos.
Area of Science:
- Developmental Biology
- Cell Biology
- Biochemistry
Background:
- Rho-associated coiled-coil-containing protein kinase (ROCK) signaling regulates actin-cytoskeleton assembly and cell contractility, crucial for embryonic development.
- Somite formation is essential for embryogenesis, involving cytoskeletal rearrangements and morphological changes.
- Inhibition of ROCK signaling has been linked to developmental defects, including ventral body wall defects (VBWD).
Purpose of the Study:
- To investigate the impact of Y-27632, a ROCK inhibitor, on somite development in post-gastrulation chick embryos.
- To elucidate the molecular mechanisms underlying Y-27632-induced embryonic malformations.
Main Methods:
- Administration of Y-27632 to chick embryos in shell-less culture.
- Assessment of somite development, body axis morphology, and cell viability.
- Western blot analysis to confirm ROCK pathway inhibition and assess protein expression.
- Histology and RT-PCR to evaluate cell death and gene expression patterns.
Main Results:
- Y-27632 treatment resulted in a kinked longitudinal body axis in chick embryos.
- ROCK downstream signaling was impaired, evidenced by decreased phosphorylated cofilin-2 expression.
- Increased cell death was observed in somites, neural tube, and ectoderm.
- Expression of key somitogenesis factors was reduced in Y-27632-treated embryos.
Conclusions:
- Y-27632 disrupts normal somite development by inhibiting ROCK signaling.
- This disruption leads to axial kinking and embryo malformation, potentially causing ventral body wall defects.
- ROCK signaling is critical for maintaining cytoskeletal integrity during early embryogenesis.
