---
_id: '14774'
abstract:
- lang: eng
text: Morphogen gradients impart positional information to cells in a homogenous
tissue field. Fgf8a, a highly conserved growth factor, has been proposed to act
as a morphogen during zebrafish gastrulation. However, technical limitations have
so far prevented direct visualization of the endogenous Fgf8a gradient and confirmation
of its morphogenic activity. Here, we monitor Fgf8a propagation in the developing
neural plate using a CRISPR/Cas9-mediated EGFP knock-in at the endogenous fgf8a
locus. By combining sensitive imaging with single-molecule fluorescence correlation
spectroscopy, we demonstrate that Fgf8a, which is produced at the embryonic margin,
propagates by diffusion through the extracellular space and forms a graded distribution
towards the animal pole. Overlaying the Fgf8a gradient curve with expression profiles
of its downstream targets determines the precise input-output relationship of
Fgf8a-mediated patterning. Manipulation of the extracellular Fgf8a levels alters
the signaling outcome, thus establishing Fgf8a as a bona fide morphogen during
zebrafish gastrulation. Furthermore, by hindering Fgf8a diffusion, we demonstrate
that extracellular diffusion of the protein from the source is crucial for it
to achieve its morphogenic potential.
acknowledgement: "We thank members of the Brand lab, as well as Justina Stark (Ivo
Sbalzarini group, Max Planck Institute of Molecular Cell Biology and Genetics, Dresden,
Germany) for project-related discussions; Darren Gilmour (University of Zurich),
Karuna Sampath (University of Warwick) and Gokul Kesavan (Vowels Lifesciences Private
Limited, Bangalore) for comments on the manuscript; personnel of the CMCB technology
platform, TU Dresden for imaging and image analysis-related support; and Maurizio
Abbate (Technical support, Arivis) for help with image analysis. We are also grateful
to Stapornwongkul and Briscoe for commenting on a preprint version of our work (Stapornwongkul
and Briscoe, 2022).\r\nThis work was supported by the Deutsche Forschungsgemeinschaft
(BR 1746/6-2, BR 1746/11-1 and BR 1746/3 to M.B.), by a Cluster of Excellence ‘Physics
of Life’ seed grant and by institutional funds from Technische Universitat Dresden
(to M.B.). Open Access funding provided by Technische Universitat Dresden. Deposited
in PMC for immediate release."
article_number: dev201559
article_processing_charge: Yes (via OA deal)
article_type: original
author:
- first_name: Rohit K
full_name: Harish, Rohit K
id: 1bae78aa-ee0e-11ec-9b76-bc42990f409d
last_name: Harish
- first_name: Mansi
full_name: Gupta, Mansi
last_name: Gupta
- first_name: Daniela
full_name: Zöller, Daniela
last_name: Zöller
- first_name: Hella
full_name: Hartmann, Hella
last_name: Hartmann
- first_name: Ali
full_name: Gheisari, Ali
last_name: Gheisari
- first_name: Anja
full_name: Machate, Anja
last_name: Machate
- first_name: Stefan
full_name: Hans, Stefan
last_name: Hans
- first_name: Michael
full_name: Brand, Michael
last_name: Brand
citation:
ama: Harish RK, Gupta M, Zöller D, et al. Real-time monitoring of an endogenous
Fgf8a gradient attests to its role as a morphogen during zebrafish gastrulation.
Development. 2023;150(19). doi:10.1242/dev.201559
apa: Harish, R. K., Gupta, M., Zöller, D., Hartmann, H., Gheisari, A., Machate,
A., … Brand, M. (2023). Real-time monitoring of an endogenous Fgf8a gradient attests
to its role as a morphogen during zebrafish gastrulation. Development.
The Company of Biologists. https://doi.org/10.1242/dev.201559
chicago: Harish, Rohit K, Mansi Gupta, Daniela Zöller, Hella Hartmann, Ali Gheisari,
Anja Machate, Stefan Hans, and Michael Brand. “Real-Time Monitoring of an Endogenous
Fgf8a Gradient Attests to Its Role as a Morphogen during Zebrafish Gastrulation.”
Development. The Company of Biologists, 2023. https://doi.org/10.1242/dev.201559.
ieee: R. K. Harish et al., “Real-time monitoring of an endogenous Fgf8a gradient
attests to its role as a morphogen during zebrafish gastrulation,” Development,
vol. 150, no. 19. The Company of Biologists, 2023.
ista: Harish RK, Gupta M, Zöller D, Hartmann H, Gheisari A, Machate A, Hans S, Brand
M. 2023. Real-time monitoring of an endogenous Fgf8a gradient attests to its role
as a morphogen during zebrafish gastrulation. Development. 150(19), dev201559.
mla: Harish, Rohit K., et al. “Real-Time Monitoring of an Endogenous Fgf8a Gradient
Attests to Its Role as a Morphogen during Zebrafish Gastrulation.” Development,
vol. 150, no. 19, dev201559, The Company of Biologists, 2023, doi:10.1242/dev.201559.
short: R.K. Harish, M. Gupta, D. Zöller, H. Hartmann, A. Gheisari, A. Machate, S.
Hans, M. Brand, Development 150 (2023).
date_created: 2024-01-10T09:18:54Z
date_published: 2023-10-01T00:00:00Z
date_updated: 2024-01-10T12:45:25Z
day: '01'
ddc:
- '570'
department:
- _id: AnKi
doi: 10.1242/dev.201559
external_id:
isi:
- '001097449100002'
pmid:
- '37665167'
file:
- access_level: open_access
checksum: 2d6f52dc33260a9b2352b8f28374ba5f
content_type: application/pdf
creator: dernst
date_created: 2024-01-10T12:41:13Z
date_updated: 2024-01-10T12:41:13Z
file_id: '14790'
file_name: 2023_Development_Harish.pdf
file_size: 12836306
relation: main_file
success: 1
file_date_updated: 2024-01-10T12:41:13Z
has_accepted_license: '1'
intvolume: ' 150'
isi: 1
issue: '19'
keyword:
- Developmental Biology
- Molecular Biology
language:
- iso: eng
month: '10'
oa: 1
oa_version: Published Version
pmid: 1
publication: Development
publication_identifier:
eissn:
- 1477-9129
issn:
- 0950-1991
publication_status: published
publisher: The Company of Biologists
quality_controlled: '1'
status: public
title: Real-time monitoring of an endogenous Fgf8a gradient attests to its role as
a morphogen during zebrafish gastrulation
tmp:
image: /images/cc_by.png
legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
short: CC BY (4.0)
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 150
year: '2023'
...