Getting a head start: Craniofacial heterochrony in marsupials involves dynamic changes to molecular and cellular mechanisms underlying neural crest development
Posted on: 12 November 2025 , updated on: 13 November 2025
Preprint posted on 25 September 2025
Those who migrate together, develop together: Marsupial cranial neural crest cells migrate as epithelial-like sheets to form craniofacial structures, providing a potential mechanism that promotes accelerated jaw development.
Selected by Heather Pollington
Categories: developmental biology, evolutionary biology
Figure 1. Comparison of marsupial and eutheria cranial neural crest cell (NCC) migration in early craniofacial development.
Significance of study
This preprint highlights the diverse mechanisms in craniofacial development among vertebrates and describes a surprising contrast to the neural crest cell (NCC) migration pattern observed in eutherian mammals. Similar NCC migratory mechanisms have been observed in amphibians and ray-finned fish1-2, and the authors speculate that this may represent a more ancestral developmental strategy that was lost in eutherian mammals, resulting in expanded diversity in craniofacial structures. Importantly, this work emphasizes that taxonomic classification may not always suggest a similarity in developmental mechanisms.
Introduction
The evolution of the vertebrate jaw was a revolutionary adaptation. It led to a highly competitive predator-prey environment creating evolutionary pressures that expanded vertebrate craniofacial diversity. Yet, our understanding of the mechanisms that promote diverse craniofacial features remains minimal.
Craniofacial formation begins in early development when multipotent stem cells arrange into three distinct flat cell layers known as the mesoderm, endoderm, and ectoderm3. The ectoderm cell layer thickens to form the neural plate, which then folds and fuses together to form the vertebrate neural tube. Following neural tube formation, cells along the neural tube border are induced to become specialized migratory cells known as neural crest cells (NCCs) 3.
NCCs are multipotent stem cells that are promoted to undergo an epithelial-to-mesenchymal transition, allowing them to migrate along the embryo and form many embryonic structures. Cranial NCCs migration generates many craniofacial elements of the vertebrate head including the maxilla and mandible bones that make up the jaw.
NCC induction is hierarchically organized by a cascade of signaling events4. Initially, morphogens promote ectoderm patterning and initiate NCC induction at the border of the neural plate. A suite of transcription factors (TFs), including SNAIL (SNAI1 and SNAI2), promotes NCC delamination from the neuroepithelium. Expression of SOX10 (and other TFs) then facilitates an epithelial to mesenchymal transition required for migration3. Interestingly, investigations in marsupial development have revealed that NCC induction is initiated especially early, when the embryo consists of flat cell layers, well before the initiation of neural folding. This heterochrony has led the authors of this preprint to investigate the mechanism that may promote NCC induction and accelerated craniofacial development.
Key findings
Marsupial pre-migratory cranial NCCs accumulate as cell aggregates in early headfold development
To understand the mechanisms that promote heterochronic craniofacial formation, Newton and colleagues observed stage 21 fat-tailed dunnart embryos (Sminthopsis crassicaudata) for cranial NCC cell distribution. Morphologically flat embryos were found to have raised headfolds consisting of thickened cell aggregates. Headfolds were found to consist of densely packed SNAI2 and SOX10 positive cells, indicating that pre-migratory cranial NCCs initially accumulate to form densely packed populations of cell aggregates.
Marsupial cranial NCCs migrate as a collective epithelial-like sheet
Characterization of the molecular properties of cranial NCC aggregates revealed that SOX10/SNAI2 positive cells distribute into distinct subdomains within the headfolds, showing a mediolateral gradient of medial SNAI2 positive cells to laterally positioned SOX10 cells. Labeling of cell adhesion proteins revealed that pre-migratory NCC aggregates retained epithelial morphology and had not yet undergone an epithelial to mesenchyme transition. In stage 22 and 23 embryos, NCCs were observed to delaminate and initiate migration as dense SNAI2-negative/SOX10-positive cell aggregates. However, NCCs maintained cell adhesion proteins revealing that the newly migratory NCCs retained epithelial cell characteristics, suggesting marsupial NCCs migrate as epithelial-like sheets.
Future directions
This preprint provides an informative characterization of marsupial NCC development to explain how rapid development of important craniofacial features occurs. In the future, it will be important to perform functional experiments to further understand the mechanisms that promote NCC epithelial-like sheet migration. The authors speculate that this mode of NCC collective migration may promote quicker cell accumulation and differentiation to facilitate rapid jaw development. This would be an exciting and insightful avenue to explore in future work.
Questions for the authors
1. Do you plan to perform functional experiments to promote migratory NCC epithelial-like sheets in a eutherian mammal, such as mouse?
2. What do you believe the consequences of promoting accelerated craniofacial development in a mouse-model might be?
3. Can you hypothesize on what may occur if marsupial NCC epithelial-like sheets remained intact but consisted of fewer cells?
References
1. Stundl, J. et al., Migratory patterns and evolutionary plasticity of cranial neural crest cells in ray-finned fish. Dev. Biol. 467, 14-29 (2020).
2. Cerny, R. et al., Combined intrinsic and extrinsic influences pattern cranial neural crest migration and pharyngeal arch morphogenesis in axolotl. Dev. Bio. 266, 252-269 (2004).
3. Bronner, M. E. & LeDouarin, N. M. Evolution and Development of the Neural Crest: An Overview. Dev Biol 366, 2–9 (2012).
4. Simoes-Costa, M. & Bronner, M. E. Establishing neural crest identity: A gene regulatory recipe. Development 142, 242-257 (2015).
doi: https://doi.org/10.1242/prelights.42055
Read preprint
(No Ratings Yet)Sign up to customise the site to your preferences and to receive alerts
Register hereAlso in the developmental biology category:
Dosage-sensitive RBFOX2 autoregulation promotes cardiomyocyte differentiation by maturing the transcriptome
Theodora Stougiannou
Post-translational Tuning of Human Cortical Progenitor Neuronal Output
Jawdat Sandakly
Aspartate transaminases are required for blood development
Hannah Pletcher
Also in the evolutionary biology category:
A high-coverage genome from a 200,000-year-old Denisovan
AND
A global map for introgressed structural variation and selection in humans
Siddharth Singh
Dissecting Gene Regulatory Networks Governing Human Cortical Cell Fate
Manuel Lessi
Beyond venomous fangs: Uloboridae spiders have lost their venom but not their toxicity
Daniel Fernando Reyes Enríquez, Marcus Oliveira
preLists in the developmental biology category:
November in preprints – DevBio & Stem cell biology
preLighters with expertise across developmental and stem cell biology have nominated a few developmental and stem cell biology (and related) preprints posted in November they’re excited about and explain in a single paragraph why. Concise preprint highlights, prepared by the preLighter community – a quick way to spot upcoming trends, new methods and fresh ideas.
| List by | Aline Grata et al. |
October in preprints – DevBio & Stem cell biology
Each month, preLighters with expertise across developmental and stem cell biology nominate a few recent developmental and stem cell biology (and related) preprints they’re excited about and explain in a single paragraph why. Short, snappy picks from working scientists — a quick way to spot fresh ideas, bold methods and papers worth reading in full. These preprints can all be found in the October preprint list published on the Node.
| List by | Deevitha Balasubramanian et al. |
October in preprints – Cell biology edition
Different preLighters, with expertise across cell biology, have worked together to create this preprint reading list for researchers with an interest in cell biology. This month, most picks fall under (1) Cell organelles and organisation, followed by (2) Mechanosignaling and mechanotransduction, (3) Cell cycle and division and (4) Cell migration
| List by | Matthew Davies et al. |
June in preprints – the CellBio edition
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. This month, categories include: (1) Cell organelles and organisation (2) Cell signaling and mechanosensation (3) Genetics/gene expression (4) Biochemistry (5) Cytoskeleton
| List by | Barbora Knotkova et al. |
Keystone Symposium – Metabolic and Nutritional Control of Development and Cell Fate
This preList contains preprints discussed during the Metabolic and Nutritional Control of Development and Cell Fate Keystone Symposia. This conference was organized by Lydia Finley and Ralph J. DeBerardinis and held in the Wylie Center and Tupper Manor at Endicott College, Beverly, MA, United States from May 7th to 9th 2025. This meeting marked the first in-person gathering of leading researchers exploring how metabolism influences development, including processes like cell fate, tissue patterning, and organ function, through nutrient availability and metabolic regulation. By integrating modern metabolic tools with genetic and epidemiological insights across model organisms, this event highlighted key mechanisms and identified open questions to advance the emerging field of developmental metabolism.
| List by | Virginia Savy, Martin Estermann |
Biologists @ 100 conference preList
This preList aims to capture all preprints being discussed at the Biologists @100 conference in Liverpool, UK, either as part of the poster sessions or the (flash/short/full-length) talks.
| List by | Reinier Prosee, Jonathan Townson |
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 |
Also in the evolutionary biology category:
November in preprints – DevBio & Stem cell biology
preLighters with expertise across developmental and stem cell biology have nominated a few developmental and stem cell biology (and related) preprints posted in November they’re excited about and explain in a single paragraph why. Concise preprint highlights, prepared by the preLighter community – a quick way to spot upcoming trends, new methods and fresh ideas.
| List by | Aline Grata et al. |
October in preprints – DevBio & Stem cell biology
Each month, preLighters with expertise across developmental and stem cell biology nominate a few recent developmental and stem cell biology (and related) preprints they’re excited about and explain in a single paragraph why. Short, snappy picks from working scientists — a quick way to spot fresh ideas, bold methods and papers worth reading in full. These preprints can all be found in the October preprint list published on the Node.
| List by | Deevitha Balasubramanian et al. |
October in preprints – Cell biology edition
Different preLighters, with expertise across cell biology, have worked together to create this preprint reading list for researchers with an interest in cell biology. This month, most picks fall under (1) Cell organelles and organisation, followed by (2) Mechanosignaling and mechanotransduction, (3) Cell cycle and division and (4) Cell migration
| List by | Matthew Davies et al. |
Biologists @ 100 conference preList
This preList aims to capture all preprints being discussed at the Biologists @100 conference in Liverpool, UK, either as part of the poster sessions or the (flash/short/full-length) talks.
| List by | Reinier Prosee, Jonathan Townson |
‘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 |
EMBO | EMBL Symposium: The organism and its environment
This preList contains preprints discussed during the 'EMBO | EMBL Symposium: The organism and its environment', organised at EMBL Heidelberg, Germany (May 2023).
| List by | Girish Kale |
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 |
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 |
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 |
TAGC 2020
Preprints recently presented at the virtual Allied Genetics Conference, April 22-26, 2020. #TAGC20
| List by | Maiko Kitaoka et al. |
ECFG15 – Fungal biology
Preprints presented at 15th European Conference on Fungal Genetics 17-20 February 2020 Rome
| List by | Hiral Shah |
COVID-19 / SARS-CoV-2 preprints
List of important preprints dealing with the ongoing coronavirus outbreak. See http://covidpreprints.com for additional resources and timeline, and https://connect.biorxiv.org/relate/content/181 for full list of bioRxiv and medRxiv preprints on this topic
| List by | Dey Lab, Zhang-He Goh |
1
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 |
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 |