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High Resolution Whole Mount In Situ Hybridization within Zebrafish Embryos to Study Gene Expression and Function
Published on: October 19, 2013
The zebrafish bcl-2 homologue Nrz controls development during somitogenesis and gastrulation via apoptosis-dependent
E Arnaud1, K F Ferri, J Thibaut
1Apoptosis and Oncogenesis Laboratory, IBCP, UMR 5086 CNRS-UCBL, IFR 128, Lyon France.
Abstract:
Although the role of the b-cell lymphoma (Bcl)-2 family of apoptosis inhibitors is well documented in tumor cells and tissue morphogenesis, their role during the early development of vertebrates is unknown. Here, we characterize Nrz, a new Bcl-2-related inhibitor of apoptosis in zebrafish. Nrz is a mitochondrial protein, antagonizing the death-accelerator Bax. The nrz gene is mainly expressed during gastrulation and somitogenesis. The knockdown of nrz with antisense morpholinos leads to alterations of the somites, correlated with an increase in apoptosis. In addition, earlier during development, in the zebrafish gastrula, nrz knockdown results in an increase of snail-1 expression at the margin and frequent gastrulation arrest at the shield stage, independently of apoptosis. Together these data suggest that Nrz, in addition to its effect on apoptosis, contributes to cell movements during gastrulation by negatively regulating the expression of Snail-1, a transcription factor that controls cell adhesion.
Insights
We discovered Nrz, a new apoptosis inhibitor in zebrafish, crucial for early vertebrate development. Nrz regulates cell movements during gastrulation and somite formation by influencing apoptosis and Snail-1 expression.
Area of Science:
- Developmental Biology
- Cell Biology
- Molecular Biology
Background:
- The Bcl-2 family of apoptosis inhibitors is vital in tumor cells and tissue development.
- Their function in early vertebrate development remains largely uncharacterized.
Purpose of the Study:
- To characterize Nrz, a novel Bcl-2-related apoptosis inhibitor in zebrafish.
- To investigate the role of Nrz during early vertebrate development, focusing on gastrulation and somitogenesis.
Main Methods:
- Utilized antisense morpholinos for Nrz knockdown in zebrafish embryos.
- Analyzed effects on apoptosis, somite formation, gastrulation, and Snail-1 expression.
Main Results:
- Nrz knockdown caused somite defects and increased apoptosis.
- Nrz knockdown also led to increased Snail-1 expression and gastrulation arrest, independent of apoptosis.
- Nrz is a mitochondrial protein that antagonizes Bax.
Conclusions:
- Nrz plays a dual role in early zebrafish development, regulating both apoptosis and cell movements during gastrulation.
- Nrz negatively regulates Snail-1 expression, impacting cell adhesion and gastrulation.
- Nrz is essential for proper somite formation and gastrulation progression.
