Close

Notch is Required for Neural Progenitor Proliferation During Embryonic Eye Regrowth

Dylan J. Guerin, Belen Gutierrez, Baoyi Zhang, Kelly Ai-Sun Tseng

Posted on: 10 March 2024 , updated on: 12 March 2024

Preprint posted on 2 February 2024

The role of a conserved cell signaling pathway in regulating the proliferation of neural progenitors during embryonic eye regrowth in a frog.

Selected by Mina Basily

Introduction:

The Notch pathway is a conserved cell signaling system in most metazoans, playing a quintessential role in development and regeneration. Notch is a cell surface receptor that interacts with membrane-bound ligands on neighboring cells, regulating cell fate, differentiation, proliferation, and apoptosis pathways [1, 2].

In this preprint, the authors explore the significance of Notch signaling, and specifically Notch1 signaling, in embryonic eye regrowth and its role in inducing retinal progenitor cell proliferation in response to injury.

For this study, African clawed frog (Xenopus laevis) embryos at developmental stage 27 (st. 27) were chosen as a suitable animal model to perform the eye ablation surgery to investigate the role of notch signaling in embryonic eye regrowth. The suitability of Xenopus embryos relies on the following features: the eye can completely be regrown in five days, and this regrowth can occur simultaneously with normal eye development on the uninjured side.

Major findings:

The role of Notch signaling in regrowing a full-size embryonic eye

To investigate the possible role of Notch1 in Xenopus embryonic eye regrowth, the authors chemically inhibited Notch signaling after an eye ablation procedure. The chemical inhibition of Notch signaling was achieved by MG132 and DAPT treatment, and this inhibition remarkably reduced the size of the regrown embryonic eye compared with the control.

To avoid any overlap in the results, and to make sure that their observations were due to the lack of Notch signaling, the authors resorted to a molecular approach to inhibit Notch signaling; they used two fluorescein morpholinos one of which is a verified morpholino against Xenopus Notch1 mRNA, and the other is a control morpholino. Each of the two morpholinos (MO) was injected separately into the left dorsal blastomere of four-cell embryos at a concentration that does not affect embryogenesis. The Notch1 morpholino reduced the size of the regrown embryonic eye in a way that resembled the effect of the chemical inhibition approach used earlier.

To confirm that the Notch1 morpholino results were due to the absence of Notch1, the preprint authors co-injected dexamethasone-inducible Notch intracellular domain (NICD) mRNA (NICD is an active form of Notch) along with the Notch1 morpholino into four-cell embryos. After eye ablation, they then injected dexamethasone to induce the activity of NICD in the presence of Notch1 morpholino; this restored full eye regrowth rates to control levels.

Revealing a critical time-window for Notch signaling during the eye regrowth

The authors’ previous experiments revealed that 87% of eye size was restored within the first two days of regrowth, which led them to suggest that maybe Notch signaling is required the most during this initial regrowth period. To investigate this hypothesis, they exposed the embryos to an MG132 inhibitor after an eye ablation at different time points within five days after the surgery. They found that the inhibition of Notch signaling during the first day (day 0) alone matched the effect of exposure to the inhibitor for five consecutive days (day 0–5), and any attempt to use the inhibitor after passing the first 24 hours post-surgery failed and the size of the regrown eye wasn’t affected (Figure 1).

Figure 1: (A) The study compares regrown eyes after MG132 treatment for various durations from surgery, (B) Graphical representation of percent of the population achieving full regrowth at 5 dps, * = p<0.01.

The role of Notch signaling in retinal differentiation and proliferation

To investigate the effect of Notch signaling on cell differentiation, three groups of embryos were fixed 3 days post-surgery: one was exposed to MG123, one was injected with Notch1 morpholinos, and one was the control. All the three groups were stained with an anti-Islet1 antibody as a marker of ganglion cells and an anti-Rhodopsin antibody as a marker of rod photoreceptor cells, one at a time. The three groups showed similar retinal patterns in terms of morphology and cell positioning with size. This means that Notch inhibition doesn’t affect retinal differentiation. To investigate the effect of Notch inhibition on cell proliferation, embryos were treated with MG123 or Notch1 morpholinos. This was followed by eye ablation surgery, after which the embryos were fixed, sectioned, and stained with (H3P) antibody, a mitosis marker. The results showed a dramatic decrease in H3P within Notch-inhibited cells compared with the control, which indicates the important role of Notch in cell proliferation.

Overexpression of Notch1 restores the ability of embryonic eye regrowth during V-ATPase inhibition

The inhibition of Notch resembled the phenotypes observed when the proton pump V-ATPase is inhibited. This proton pump is considered an important player in allowing eye regrowth but not development, which is why the authors wanted to investigate if Notch and V-ATPase could interact to regulate eye regrowth. To investigate this, they injected four-cell embryos with Dex-inducible NICD mRNA along with Concanamycin A,a V-ATPase inhibitor, after eye ablation. Another control group was treated with Concanamycin alone. The first group was able to regrow eyes in 75.5% of cases, whereas the control group was only able to regrow eyes in 22.6% of cases. This highlights the importance of the ectopic activation of Notch in rescuing eye growth and the fact that it acts downstream ofV-ATPase.

Why I chose this preprint and what I liked about it:

This preprint describes a novel role for Notch signaling during eye regrowth. Understanding the different aspects that govern regrowth and regeneration in animals such as Xenopus will contribute to solving the complex riddles faced when studying regeneration in higher vertebrates. Also, this study allows us to better appreciate how important Notch signaling is in processes such as proliferation and neural regrowth. I particularly liked how the authors have used different inhibitory approaches to confirm the role of Notch, and how they managed to describe the relationship between Notch and V-ATPase in allowing eye regrowth.

Questions for the authors:

  • Do you have any idea if Notch inhibition affects Pax6 in any way during the regrowth?
  • Would you consider, in the future, any molecular study to compare the role of Notch in regulating cell proliferation and differentiation between eye regrowth and eye development?
  • What is the future of this project?

References:

  1. Hori K, Sen A, Artavanis-Tsakonas S. Notch signaling at a glance. J Cell Sci. 2013 May 15;126(Pt 10):2135-40. doi: 10.1242/jcs.127308. Epub 2013 May 31. PMID: 23729744; PMCID: PMC3672934.
  2. Gao J, Fan L, Zhao L, Su Y. The interaction of Notch and Wnt signaling pathways in vertebrate regeneration. Cell Regen. 2021 Apr 1;10(1):11. doi: 10.1186/s13619-020-00072-2. PMID: 33791915; PMCID: PMC8012441.

Tags: cell communication, cell signaling, conserved cell-cell interaction mechanisms, embryos, frog, notch, regrowth and regeneration

doi: https://doi.org/10.1242/prelights.36762

Read preprint (No Ratings Yet)

Have your say

Your email address will not be published. Required fields are marked *

This site uses Akismet to reduce spam. Learn how your comment data is processed.

Sign up to customise the site to your preferences and to receive alerts

Register here

Also in the cell biology category:

Platelet-derived LPA16:0 inhibits adult neurogenesis and stress resilience in anxiety disorder

Thomas Larrieu, Charline Carron, Fabio Grieco, et al.

Selected by 04 December 2024

Harvey Roweth

Neuroscience

Investigating Mechanically Activated Currents from Trigeminal Neurons of Non-Human Primates

Karen A Lindquist, Jennifer Mecklenburg, Anahit H. Hovhannisyan, et al.

Selected by 04 December 2024

Vanessa Ehlers

Neuroscience

Motor Clustering Enhances Kinesin-driven Vesicle Transport

Rui Jiang, Qingzhou Feng, Daguan Nong, et al.

Selected by 16 November 2024

Sharvari Pitke

Biophysics

Also in the developmental biology category:

Cellular signalling protrusions enable dynamic distant contacts in spinal cord neurogenesis

Joshua Hawley, Robert Lea, Veronica Biga, et al.

Selected by 15 November 2024

Ankita Walvekar

Developmental Biology

Actin-based deformations of the nucleus control multiciliated ependymal cell differentiation

Marianne Basso, Alexia Mahuzier, Syed Kaabir Ali, et al.

Selected by 30 October 2024

Ryan Harrison

Developmental Biology

HIF1A contributes to the survival of aneuploid and mosaic pre-implantation embryos

Estefania Sanchez-Vasquez, Marianne E. Bronner, Magdalena Zernicka-Goetz

Selected by 11 October 2024

Anchel De Jaime Soguero

Developmental Biology

preLists in the cell biology category:

November in preprints – the CellBio edition

This is the first community-driven preList! A group of preLighters, with expertise in different areas of cell biology, have worked together to create this preprint reading lists for researchers with an interest in cell biology. Categories include: 1) cancer cell biology 2) cell cycle and division 3) cell migration and cytoskeleton 4) cell organelles and organisation 5) cell signalling and mechanosensing 6) genetics/gene expression

 



List by Felipe Del Valle Batalla et al.

BSCB-Biochemical Society 2024 Cell Migration meeting

This preList features preprints that were discussed and presented during the BSCB-Biochemical Society 2024 Cell Migration meeting in Birmingham, UK in April 2024. Kindly put together by Sara Morais da Silva, Reviews Editor at Journal of Cell Science.

 



List by Reinier Prosee

‘In preprints’ from Development 2022-2023

A list of the preprints featured in Development's 'In preprints' articles between 2022-2023

 



List by Alex Eve, Katherine Brown

preLights peer support – preprints of interest

This is a preprint repository to organise the preprints and preLights covered through the 'preLights peer support' initiative.

 



List by preLights peer support

The Society for Developmental Biology 82nd Annual Meeting

This preList is made up of the preprints discussed during the Society for Developmental Biology 82nd Annual Meeting that took place in Chicago in July 2023.

 



List by Joyce Yu, Katherine Brown

CSHL 87th Symposium: Stem Cells

Preprints mentioned by speakers at the #CSHLsymp23

 



List by Alex Eve

Journal of Cell Science meeting ‘Imaging Cell Dynamics’

This preList highlights the preprints discussed at the JCS meeting 'Imaging Cell Dynamics'. The meeting was held from 14 - 17 May 2023 in Lisbon, Portugal and was organised by Erika Holzbaur, Jennifer Lippincott-Schwartz, Rob Parton and Michael Way.

 



List by Helen Zenner

9th International Symposium on the Biology of Vertebrate Sex Determination

This preList contains preprints discussed during the 9th International Symposium on the Biology of Vertebrate Sex Determination. This conference was held in Kona, Hawaii from April 17th to 21st 2023.

 



List by Martin Estermann

Alumni picks – preLights 5th Birthday

This preList contains preprints that were picked and highlighted by preLights Alumni - an initiative that was set up to mark preLights 5th birthday. More entries will follow throughout February and March 2023.

 



List by Sergio Menchero et al.

CellBio 2022 – An ASCB/EMBO Meeting

This preLists features preprints that were discussed and presented during the CellBio 2022 meeting in Washington, DC in December 2022.

 



List by Nadja Hümpfer et al.

Fibroblasts

The advances in fibroblast biology preList explores the recent discoveries and preprints of the fibroblast world. Get ready to immerse yourself with this list created for fibroblasts aficionados and lovers, and beyond. Here, my goal is to include preprints of fibroblast biology, heterogeneity, fate, extracellular matrix, behavior, topography, single-cell atlases, spatial transcriptomics, and their matrix!

 



List by Osvaldo Contreras

EMBL Synthetic Morphogenesis: From Gene Circuits to Tissue Architecture (2021)

A list of preprints mentioned at the #EESmorphoG virtual meeting in 2021.

 



List by Alex Eve

FENS 2020

A collection of preprints presented during the virtual meeting of the Federation of European Neuroscience Societies (FENS) in 2020

 



List by Ana Dorrego-Rivas

Planar Cell Polarity – PCP

This preList contains preprints about the latest findings on Planar Cell Polarity (PCP) in various model organisms at the molecular, cellular and tissue levels.

 



List by Ana Dorrego-Rivas

BioMalPar XVI: Biology and Pathology of the Malaria Parasite

[under construction] Preprints presented at the (fully virtual) EMBL BioMalPar XVI, 17-18 May 2020 #emblmalaria

 



List by Dey Lab, Samantha Seah

1

Cell Polarity

Recent research from the field of cell polarity is summarized in this list of preprints. It comprises of studies focusing on various forms of cell polarity ranging from epithelial polarity, planar cell polarity to front-to-rear polarity.

 



List by Yamini Ravichandran

TAGC 2020

Preprints recently presented at the virtual Allied Genetics Conference, April 22-26, 2020. #TAGC20

 



List by Maiko Kitaoka et al.

3D Gastruloids

A curated list of preprints related to Gastruloids (in vitro models of early development obtained by 3D aggregation of embryonic cells). Updated until July 2021.

 



List by Paul Gerald L. Sanchez and Stefano Vianello

ECFG15 – Fungal biology

Preprints presented at 15th European Conference on Fungal Genetics 17-20 February 2020 Rome

 



List by Hiral Shah

ASCB EMBO Annual Meeting 2019

A collection of preprints presented at the 2019 ASCB EMBO Meeting in Washington, DC (December 7-11)

 



List by Madhuja Samaddar et al.

EMBL Seeing is Believing – Imaging the Molecular Processes of Life

Preprints discussed at the 2019 edition of Seeing is Believing, at EMBL Heidelberg from the 9th-12th October 2019

 



List by Dey Lab

Autophagy

Preprints on autophagy and lysosomal degradation and its role in neurodegeneration and disease. Includes molecular mechanisms, upstream signalling and regulation as well as studies on pharmaceutical interventions to upregulate the process.

 



List by Sandra Malmgren Hill

Lung Disease and Regeneration

This preprint list compiles highlights from the field of lung biology.

 



List by Rob Hynds

Cellular metabolism

A curated list of preprints related to cellular metabolism at Biorxiv by Pablo Ranea Robles from the Prelights community. Special interest on lipid metabolism, peroxisomes and mitochondria.

 



List by Pablo Ranea Robles

BSCB/BSDB Annual Meeting 2019

Preprints presented at the BSCB/BSDB Annual Meeting 2019

 



List by Dey Lab

MitoList

This list of preprints is focused on work expanding our knowledge on mitochondria in any organism, tissue or cell type, from the normal biology to the pathology.

 



List by Sandra Franco Iborra

Biophysical Society Annual Meeting 2019

Few of the preprints that were discussed in the recent BPS annual meeting at Baltimore, USA

 



List by Joseph Jose Thottacherry

ASCB/EMBO Annual Meeting 2018

This list relates to preprints that were discussed at the recent ASCB conference.

 



List by Dey Lab, Amanda Haage

Also in the developmental biology category:

BSDB/GenSoc Spring Meeting 2024

A list of preprints highlighted at the British Society for Developmental Biology and Genetics Society joint Spring meeting 2024 at Warwick, UK.

 



List by Joyce Yu, Katherine Brown

GfE/ DSDB meeting 2024

This preList highlights the preprints discussed at the 2024 joint German and Dutch developmental biology societies meeting that took place in March 2024 in Osnabrück, Germany.

 



List by Joyce Yu

‘In preprints’ from Development 2022-2023

A list of the preprints featured in Development's 'In preprints' articles between 2022-2023

 



List by Alex Eve, Katherine Brown

preLights peer support – preprints of interest

This is a preprint repository to organise the preprints and preLights covered through the 'preLights peer support' initiative.

 



List by preLights peer support

The Society for Developmental Biology 82nd Annual Meeting

This preList is made up of the preprints discussed during the Society for Developmental Biology 82nd Annual Meeting that took place in Chicago in July 2023.

 



List by Joyce Yu, Katherine Brown

CSHL 87th Symposium: Stem Cells

Preprints mentioned by speakers at the #CSHLsymp23

 



List by Alex Eve

Journal of Cell Science meeting ‘Imaging Cell Dynamics’

This preList highlights the preprints discussed at the JCS meeting 'Imaging Cell Dynamics'. The meeting was held from 14 - 17 May 2023 in Lisbon, Portugal and was organised by Erika Holzbaur, Jennifer Lippincott-Schwartz, Rob Parton and Michael Way.

 



List by Helen Zenner

9th International Symposium on the Biology of Vertebrate Sex Determination

This preList contains preprints discussed during the 9th International Symposium on the Biology of Vertebrate Sex Determination. This conference was held in Kona, Hawaii from April 17th to 21st 2023.

 



List by Martin Estermann

Alumni picks – preLights 5th Birthday

This preList contains preprints that were picked and highlighted by preLights Alumni - an initiative that was set up to mark preLights 5th birthday. More entries will follow throughout February and March 2023.

 



List by Sergio Menchero et al.

CellBio 2022 – An ASCB/EMBO Meeting

This preLists features preprints that were discussed and presented during the CellBio 2022 meeting in Washington, DC in December 2022.

 



List by Nadja Hümpfer et al.

2nd Conference of the Visegrád Group Society for Developmental Biology

Preprints from the 2nd Conference of the Visegrád Group Society for Developmental Biology (2-5 September, 2021, Szeged, Hungary)

 



List by Nándor Lipták

Fibroblasts

The advances in fibroblast biology preList explores the recent discoveries and preprints of the fibroblast world. Get ready to immerse yourself with this list created for fibroblasts aficionados and lovers, and beyond. Here, my goal is to include preprints of fibroblast biology, heterogeneity, fate, extracellular matrix, behavior, topography, single-cell atlases, spatial transcriptomics, and their matrix!

 



List by Osvaldo Contreras

EMBL Synthetic Morphogenesis: From Gene Circuits to Tissue Architecture (2021)

A list of preprints mentioned at the #EESmorphoG virtual meeting in 2021.

 



List by Alex Eve

EMBL Conference: From functional genomics to systems biology

Preprints presented at the virtual EMBL conference "from functional genomics and systems biology", 16-19 November 2020

 



List by Jesus Victorino

Single Cell Biology 2020

A list of preprints mentioned at the Wellcome Genome Campus Single Cell Biology 2020 meeting.

 



List by Alex Eve

Society for Developmental Biology 79th Annual Meeting

Preprints at SDB 2020

 



List by Irepan Salvador-Martinez, Martin Estermann

FENS 2020

A collection of preprints presented during the virtual meeting of the Federation of European Neuroscience Societies (FENS) in 2020

 



List by Ana Dorrego-Rivas

Planar Cell Polarity – PCP

This preList contains preprints about the latest findings on Planar Cell Polarity (PCP) in various model organisms at the molecular, cellular and tissue levels.

 



List by Ana Dorrego-Rivas

Cell Polarity

Recent research from the field of cell polarity is summarized in this list of preprints. It comprises of studies focusing on various forms of cell polarity ranging from epithelial polarity, planar cell polarity to front-to-rear polarity.

 



List by Yamini Ravichandran

TAGC 2020

Preprints recently presented at the virtual Allied Genetics Conference, April 22-26, 2020. #TAGC20

 



List by Maiko Kitaoka et al.

3D Gastruloids

A curated list of preprints related to Gastruloids (in vitro models of early development obtained by 3D aggregation of embryonic cells). Updated until July 2021.

 



List by Paul Gerald L. Sanchez and Stefano Vianello

ASCB EMBO Annual Meeting 2019

A collection of preprints presented at the 2019 ASCB EMBO Meeting in Washington, DC (December 7-11)

 



List by Madhuja Samaddar et al.

EDBC Alicante 2019

Preprints presented at the European Developmental Biology Congress (EDBC) in Alicante, October 23-26 2019.

 



List by Sergio Menchero et al.

EMBL Seeing is Believing – Imaging the Molecular Processes of Life

Preprints discussed at the 2019 edition of Seeing is Believing, at EMBL Heidelberg from the 9th-12th October 2019

 



List by Dey Lab

SDB 78th Annual Meeting 2019

A curation of the preprints presented at the SDB meeting in Boston, July 26-30 2019. The preList will be updated throughout the duration of the meeting.

 



List by Alex Eve

Lung Disease and Regeneration

This preprint list compiles highlights from the field of lung biology.

 



List by Rob Hynds

Young Embryologist Network Conference 2019

Preprints presented at the Young Embryologist Network 2019 conference, 13 May, The Francis Crick Institute, London

 



List by Alex Eve

Pattern formation during development

The aim of this preList is to integrate results about the mechanisms that govern patterning during development, from genes implicated in the processes to theoritical models of pattern formation in nature.

 



List by Alexa Sadier

BSCB/BSDB Annual Meeting 2019

Preprints presented at the BSCB/BSDB Annual Meeting 2019

 



List by Dey Lab

Zebrafish immunology

A compilation of cutting-edge research that uses the zebrafish as a model system to elucidate novel immunological mechanisms in health and disease.

 



List by Shikha Nayar
Close