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Updated: May 24, 2026
High Resolution Whole Mount In Situ Hybridization within Zebrafish Embryos to Study Gene Expression and Function
Published on: October 19, 2013
Zebrafish arl6ip1 is required for neural crest development during embryogenesis
Chi-Tang Tu1, Tzu-Ching Yang, Hsing-Yen Huang
1Institute of Molecular and Cellular Biology, National Taiwan University, Taipei, Taiwan.
Background:
Although the embryonic expression pattern of ADP ribosylation factor-like 6 interacting protein 1 (Arl6ip1) has been reported, its function in neural crest development is unclear.
Methods/Principal Findings:
We found that knockdown of Arl6ip1 caused defective embryonic neural crest derivatives that were particularly severe in craniofacial cartilages. Expressions of the ectodermal patterning factors msxb, dlx3b, and pax3 were normal, but the expressions of the neural crest specifier genes foxd3, snai1b, and sox10 were greatly reduced. These findings suggest that arl6ip1 is essential for specification of neural crest derivatives, but not neural crest induction. Furthermore, we revealed that the streams of crestin- and sox10-expressing neural crest cells, which migrate ventrally from neural tube into trunk, were disrupted in arl6ip1 morphants. This migration defect was not only in the trunk neural crest, but also in the enteric tract where the vagal-derived neural crest cells failed to populate the enteric nervous system. We found that this migration defect was induced by dampened Shh signaling, which may have resulted from defective cilia. These data further suggested that arl6ip1 is required for neural crest migration. Finally, by double-staining of TUNEL and crestin, we confirmed that the loss of neural crest cells could not be attributed to apoptosis.
Conclusions/Significance:
Therefore, we concluded that arl6ip1 is required for neural crest migration and sublineage specification.
Insights
ADP ribosylation factor-like 6 interacting protein 1 (Arl6ip1) is essential for neural crest cell migration and specification. Its absence disrupts craniofacial development and enteric nervous system formation by impairing cell movement.
Area of Science:
- Developmental Biology
- Molecular Biology
- Genetics
Background:
- The role of ADP ribosylation factor-like 6 interacting protein 1 (Arl6ip1) in embryonic development, particularly neural crest formation, remains largely unknown.
- Previous studies have documented the expression patterns of Arl6ip1 during embryogenesis.
Purpose of the Study:
- To investigate the function of Arl6ip1 in neural crest development and its contribution to craniofacial and enteric nervous system formation.
- To elucidate the molecular mechanisms underlying Arl6ip1's role in neural crest cell specification and migration.
Main Methods:
- Knockdown of Arl6ip1 using morpholino-based approaches in zebrafish embryos.
- Analysis of neural crest derivative formation and gene expression patterns (msxb, dlx3b, pax3, foxd3, snai1b, sox10).
- Assessment of neural crest cell migration in trunk and vagal neural crest populations.
- Investigation of Shh signaling pathway and cilia function.
- TUNEL staining to evaluate apoptosis.
Main Results:
- Arl6ip1 knockdown led to severe defects in craniofacial cartilage development and reduced expression of neural crest specifier genes (foxd3, snai1b, sox10).
- Neural crest cell migration was significantly impaired in both trunk and enteric nervous system, affecting vagal neural crest colonization.
- Defective Arl6ip1 function was linked to dampened Shh signaling, potentially due to ciliary dysfunction, and did not result from increased apoptosis.
Conclusions:
- Arl6ip1 plays a critical role in neural crest cell migration and the specification of neural crest sublineages.
- The findings highlight Arl6ip1's importance in craniofacial development and the proper formation of the enteric nervous system.
- Arl6ip1 is essential for maintaining Shh signaling, which is crucial for neural crest cell movement.
