# Cell lineage and transcriptional analysis of the vertebrate neural plate border

> **NIH NIH R01** · CALIFORNIA INSTITUTE OF TECHNOLOGY · 2021 · $523,810

## Abstract

Bronner, M.E.
 In the early embryo, the neural plate border region contributes to diverse cell fates, ranging from neural
crest cells and ectodermal placode cells to neurons of the central nervous system. Despite extensive studies of
the neural crest and ectodermal placodes at post-neurula stages, surprisingly little is known about how these
populations become distinct from one another within the early neural plate border. Based on our preliminary
data, we hypothesize that many neural plate border cells are multipotent as evidenced by their
concomitant expression of markers characteristic of several fates. We will test this hypothesis by: 1)
conducting a detailed analysis of the emerging neural plate border region by multiplex protein and gene
expression profiling coupled with cell lineage analysis and 2) examining how perturbation of transcription factor
levels affects expression profiles and lineage allocations of individual neural plate border cells. The significance
of this proposal is that it will be the first to test how and when ectodermal placode precursors are segregated
from neural crest and neural precursors at the neural plate border. The following aims will be performed:
Aim 1: High resolution analysis of protein expression of neural plate, neural crest, placode and other
 ectodermal markers in the neural plate border as a function of time. We will examine co-expression of
 transcription factors associated with neural crest, placode, neural plate and ectodermal lineages
 quantitatively and at single cell resolution in chick gastrula to neurula stages to determine their degree of
 overlap and if/when a discrete separation occurs between them in the neural plate border. To take this to a
 multiplex level, we will then perform single molecule fluorescent in situ hybridization analysis (smFISH) at
 similar stages with 35 or more probes selected from known genes and new candidates from our single cell
 RNA-seq dataset.
Aim 2: Molecular dissection of regulatory interactions that mediate gene expression and cell fate
 choice at the neural plate border. We will examine the consequence of perturbing individual transcription
 factors (e.g. Pax7, Sox2, Six1) on expression of others neural plate border genes at the population and
 single cell level. To examine inputs that regulate neural plate border formation, we will dissect novel
 enhancers for Pax7, Six1 and other genes to determine direct regulatory inputs. Finally, we will examine
 how balancing levels of transcription factors may influence other factors in the neural plate border region.
Aim 3: Single cell lineage analysis of cells at the neural plate border. To definitively test whether
 individual cells at the neural plate border have restricted or broad developmental potential, we will carry out
 single cell lineage analysis by performing iontophoretic injection of lysinated rhodamine dextran into
 individual neural plate border cells. We also will use enhancers for Pax7, Sox2, or Six1...

## Key facts

- **NIH application ID:** 10085629
- **Project number:** 5R01DE027538-04
- **Recipient organization:** CALIFORNIA INSTITUTE OF TECHNOLOGY
- **Principal Investigator:** Marianne Bronner
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $523,810
- **Award type:** 5
- **Project period:** 2018-02-01 → 2023-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10085629

## Citation

> US National Institutes of Health, RePORTER application 10085629, Cell lineage and transcriptional analysis of the vertebrate neural plate border (5R01DE027538-04). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10085629. Licensed CC0.

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