# Clonal analysis of the cranial neural crest

> **NIH NIH R01** · CALIFORNIA INSTITUTE OF TECHNOLOGY · 2020 · $487,253

## Abstract

A major question in developmental biology is how precursor cells give rise to diverse sets
of differentiated cell types. This proposal tackles the question of multipotency and migratory
behavior of neural crest cells, focusing on the cranial neural crest due to its broad ability to
contribute to numerous and diverse cell types, as distinct as neurons and cartilage. Although
classical grafting experiments have elucidated the derivatives of the neural crest, comparatively
little is known about the developmental potential of individual cranial neural crest cells in vivo.
Here, we propose to use replication incompetent avian retroviruses encoding different
fluorescent fluorophores to label dorsal neural tubes in order to perform clonal analyses.
The goal is to examine the developmental potential, movement and morphogenesis of
individual or small populations of cranial neural crest cells. Experiments will be performed
on avian embryos because of several advantages. Chick embryos are easily accessible to
retroviral infection and experimental perturbation at early stages of development, allowing
temporally and spatially controlled manipulation. Birds like humans are amniotes but, unlike mice,
develop outside the mother. Therefore, they are much more accessible at early stage, while
developing in a manner that is morphologically nearly identical to human embryos at comparable
stages.
Aim 1: Retrovirally mediated clonal analysis of the chick cranial neural crest: The cranial
neural tube of chick embryos will be infected with replication incompetent avian retroviruses that
encode four different fluorophores. Clonality will be established by visual observation of single
cells a few hours after infection. We will then follow the long term fate of clonally related cells as
a function of time by examining their localization and differentiation using antibody markers
characteristic of various cell fates.
Aim 2: Coupling lineage analysis with single molecule Fluorescent In Situ Hybridization
to examine multiplex gene expression of clonally related cells. We will couple lineage
analysis with a novel adaptation of smFISH that we have recently developed that allows multiplex
analysis of gene expression at single cell resolution. Spatial Genomic Analysis (SGA) enables
simultaneous analysis of the expression of 35 or more genes on tissue sections at migratory and
post-migratory stages. We will combine clonal analysis with SGA to determine the genes co-
expressed by clonally related cells using markers of various lineages together with neural crest
and pluripotency genes to characterize the transcriptional profile of clonally related genes.
Aim 3: Analysis of migratory interactions between clonally related cells: We will examine
the migratory behavior of clonally related cells both in whole mount, using in ovo imaging, as well
as in slice tissue sections to visualize interactions between sister cells and unrelated neighbors.
Once normal migratory patterns and cell int...

## Key facts

- **NIH application ID:** 9836624
- **Project number:** 5R01DE027568-03
- **Recipient organization:** CALIFORNIA INSTITUTE OF TECHNOLOGY
- **Principal Investigator:** Marianne Bronner
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $487,253
- **Award type:** 5
- **Project period:** 2018-01-01 → 2022-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9836624

## Citation

> US National Institutes of Health, RePORTER application 9836624, Clonal analysis of the cranial neural crest (5R01DE027568-03). Retrieved via AI Analytics 2026-05-21 from https://api.ai-analytics.org/grant/nih/9836624. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*