# Translational regulation of limb bud initiation

> **NIH NIH R21** · UNIVERSITY OF TEXAS AT AUSTIN · 2022 · $198,125

## Abstract

Gene regulatory networks underlying most common developmental systems have been characterized in
increasingly granular detail using mRNA expression as a primary readout. Yet, mRNA expression levels are
poorly correlated with protein levels. A large component of the unexplained variation in protein levels is thought
to be attributable to translation efficiency, the number of proteins synthesized per mRNA. Translationally
regulated genes can be obtained by identifying actively translated mRNAs by ribosome profiling. In
combination with conventional gene expression techniques, this approach provides a method for calculating
the translational efficiency of mRNAs. However, this approach requires large numbers of cells (more than 1
million) that are unfeasible for application to most in vivo developmental systems. One of us recently pioneered
the development of a novel microfluidic approach for ribosome profiling termed Ribo-ITP that delivers high
coverage and high-resolution ribosome occupancy measurements from ultra-low input samples (~100 cells).
This method enables the identification of translationally regulated genes even in dynamic developmental
systems with limited cellular material. We propose to apply this new approach to determine the mechanisms
regulating the initial phase of limb outgrowth. This process is mediated by FGF10, which is essential for the
epithelial to mesenchymal transformation (EMT) of the lateral plate mesoderm as well as for subsequent limb
bud outgrowth. We hypothesize that limb bud EMT and FGF10-responsive gene regulatory networks are
translationally regulated. In specific aim 1, we will determine the contribution of translational regulation to limb
bud epithelial to mesenchymal transformation (EMT). In specific aim 2, we will identify transcriptional and
translational networks mediated by FGF10, which is necessary and sufficient for mediating limb bud initiation
via poorly understood mechanisms. At the completion of these studies, we will have characterized the process
of EMT in limb bud initiation and determined the contribution of translational control to this process.
Furthermore, we will have identified the FGF10-mediated transcriptional network of genes, and determined if
they are translationally regulated, substantially advancing our understanding of limb bud initiation. Collectively,
this proposal has the potential to have a transformative impact on our understanding of how translational
regulation influences vertebrate organogenesis.

## Key facts

- **NIH application ID:** 10538940
- **Project number:** 1R21HD110096-01
- **Recipient organization:** UNIVERSITY OF TEXAS AT AUSTIN
- **Principal Investigator:** Can Cenik
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $198,125
- **Award type:** 1
- **Project period:** 2022-08-19 → 2024-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10538940

## Citation

> US National Institutes of Health, RePORTER application 10538940, Translational regulation of limb bud initiation (1R21HD110096-01). Retrieved via AI Analytics 2026-05-27 from https://api.ai-analytics.org/grant/nih/10538940. Licensed CC0.

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