bioRxiv Science⌕ Search
Home/Archive/bioRxiv

bioRxiv · 10.64898/2026.11.03.763904

Physiological caffeine concentrations do not affect enteric neural crest cell migration in a mouse model

Romito, O.·Peres, J.·Dicu, M.·Lelong, E.·Le Parco, I.·Bondurand, N.·Chevalier, N. R.
View original source ↗Export metadata ↓

Abstract

The defect in enteric neural crest cell (ENCCs) migration that causes Hirschsprung disease (HD) in newborns is linked to genetic and likely also environmental factors. Among these, maternal coffee consumption has been associated with increased HD risk, yet the role of caffeine on ENCC migration has remained unexplored. Here, we show that although mouse ENCCs express caffeine-sensitive ADORA2A receptors, physiological doses of caffeine do not perturb endogenous ENCC calcium activity. We find that caffeine can modulate ENCC outgrowth in a 2D culture model but that it does not affect migration in a 3D collagen gel and in the ex-vivo embryonic gut. We conclude that physiological doses of caffeine do not hamper ENCC migration in the ex-vivo embryonic mouse intestine, thereby discarding a potential direct environmental mechanism of caffeine in HD.

Explore related subjects

developmental biologyPreprint

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Romito, O., Peres, J., Dicu, M., Lelong, E., Le Parco, I., Bondurand, N., Chevalier, N. R.. 2026-11-04. Physiological caffeine concentrations do not affect enteric neural crest cell migration in a mouse model. https://doi.org/10.64898/2026.11.03.763904

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

bioRxiv2026

PPI-Rescue: restoration of eye development via functional reconstitution of a transcription complex.

The activity of transcription factors (TFs) can be modulated through protein-protein interactions (PPIs) with cofactors which can serve to enhance, repress or alter the specificity of TF function. Current genetic and molecular approaches such as knockout and overexpression studies are likely to affect all possible activities of a TF, presenting a dilemma when attempting to parse out how TFs function within a cell. Hence, there is a need to devise a technique that will allow for mechanistic insight to be gained into the function(s) of discrete transcription complexes. Herein we report ''PPI-Rescue'', an approach that leverages bespoke mutations in TFs and the selective reconstitution of specific complexes to study the molecular properties and biological functions of discrete transcription complexes in vitro and in vivo. We demonstrate PPI-Rescue using the conserved Drosophila transcription factor Sine oculis (So) and cofactor Eyes Absent (Eya). To validate this approach, we engineered a missense mutation, So{Delta}Eya (C110S), that selectively abrogates PPI-Eya to show that PPI-Eya is necessary for the transcriptional activity of So in cultured cells and to promote eye formation in Drosophila. Using heterologous leucine zipper domains, we specifically reconstituted the So{Delta}Eya-Eya PPI and show that this complex is transcriptionally active in cells and can rescue eye development. PPI-Rescue was also used to restore transcriptional activity to So{Delta}PPI (V111E), an allele that mimics the human SIX1(V17E) allele associated with Branchio-Oto-Renal syndrome. PPI-Rescue thus represents an innovative approach to understanding the complex molecular logic of transcription complexes in both development and disease.

developmental biology↗
bioRxiv2026

Oocyte-stored Mif promotes preimplantation development as an embryonic extracellular signal

Mammalian preimplantation embryos develop more efficiently in groups than in isolation, suggesting that embryos secrete embryotrophic factors that promote development. However, the endogenous embryotrophic signals responsible for this long-recognized phenomenon remain incompletely defined. Using an unbiased proteomic screen for stable maternal proteins that may function as embryonic extracellular signals, we identified the cytokine macrophage migration inhibitory factor (Mif) as an abundant oocyte-stored protein that persists for several days after fertilization in mice. Quantitative measurements of spent culture medium reveal that developing embryos release Mif, with a substantial fraction derived from the maternal deposit. Extracellular Mif levels peak 3-4 days post fertilization, the same period when Mif knockout preimplantation embryos produce its strongest phenotype. Embryos lacking maternal, but not zygotic, Mif display defective blastocyst growth even in group culture. Individually cultured embryos exhibit similar blastocyst growth defects that can be rescued by supplementing recombinant Mif at physiologically relevant concentrations. Mif activates the PI3K-Akt-mTORC1 pathway through the cell-surface receptor Cd74 to promote blastocyst growth and cell proliferation. Together, these findings identify Mif as a potential endogenous contributor to the density-dependent embryotrophic effects and reveal that maternally stored proteins may function long after fertilization as extracellular signaling molecules to support preimplantation development in mammals.

developmental biology↗
bioRxiv2026

Single-cell transcriptomics of enriched rare neural lineages of Tribolium castaneum

Background Single-cell transcriptomic atlases have transformed our understanding of cell diversity. However, studies designed to profile entire tissues or organisms often omit or underrepresent rare cell populations. Enriching selected populations before sequencing offers a powerful approach to overcoming this limitation, but its application in insects has largely focused on Drosophila, limiting opportunities for comparative studies. Here, we establish complementary workflows for targeted transcriptomic profiling of rare neural populations in the red flour beetle, Tribolium castaneum. Results We developed two workflows combining population enrichment with single-cell or single-nucleus RNA sequencing. First, we generated cell suspensions from a transgenic reporter line labelling embryonic neuroblasts, enriched fluorescently labelled cells by fluorescence-activated cell sorting, and performed single-cell RNA sequencing. Second, we immunolabelled first-instar larval brains with an antibody against the nuclear transcription factor Rx, isolated and sorted labelled nuclei, and performed single-nucleus RNA sequencing. Both workflows yielded material suitable for transcriptomic profiling, demonstrating complementary reporter-based and antibody-based strategies for accessing selected neural populations at different developmental stages. Conclusions These workflows establish a methodological framework for targeted single-cell and single-nucleus transcriptomic studies of rare or specific neural populations in Tribolium. They provide practical starting points for adaptation to other insect systems, expanding opportunities for comparative investigations of neural development and cell-type evolution.

developmental biology↗