bioRxiv Science⌕ Search

Biology subjects

Raiola, M.

Publications and source records attributed to Raiola, M..

3 recordsLinked to original sources

Quantitative computerized analysis demonstrates strongly compartmentalized tissue deformation patterns underlying mammalian heart tube formation

The quantitative analysis of tissue deformation at cellular resolution remains an important challenge in mammalian organogenesis. Here, we developed a new computational workflow to extract regional and temporal patterns of tissue deformation and applied it to a collection of live microscopy datasets from mouse cardiogenesis. We devised a method to track tissue deformation directly from time-lapse raw images and experimentally validated the method by comparison with actual cell tracks. We then used a machine-learning approach to temporally and spatially align different specimens and reconstruct a single statistical model of tissue motion, deducing maps of strain, anisotropy, and tissue growth. We also implemented a virtual fate mapping tool that allows tracking any initial position in the cardiac primordium onto the linear heart tube. Our study reveals predominant local cellular coherence during the deformation of the cardiac tissue, whereas strong compartmentalization of tissue deformation patterns transforms the bilateral cardiac primordium into a 3D longitudinal heart tube. At the future outer curvature of the primitive tube, the ventricular chamber forms by expansion of the tissue in a hemi-barrel shape with two harnessing belts; one that constrains tissue expansion at the arterial pole and one that constrains the expansion at the venous pole. Our study provides a new approach to understanding heart morphogenesis and proposes a new model of primitive heart tube formation.

developmental biology↗

A method for analysing tissue motion and deformation during mammalian organogenesis

Understanding tissue morphogenesis is an important goal in developmental biology and tissue engineering. Accurately describing tissue deformation processes and how cell rearrangements contribute to these is a challenging task. Live analysis of morphogenesis in 3D is frequently used to obtain source data that allow to extract such features from developing organs. However, several limitations are encountered when applying these methodologies to mammalian embryos. The mouse embryo is the most frequently used model, but most studies use a very limited number of specimens and present only individual acquisitions due to constraints imposed by embryo culture and imaging. Here we used data from live analysis of embryonic heart development to develop methodologies that allow to 1) deduce tissue deformation and cell movements from image intensity flows, 2) stage and register data from several specimens/acquisitions to a dynamic consensus model of heart development, 3) statistically calculate and map to the model tissue growth and anisotropy and 4) generate a digital model of heart development that allows in-silico pseudo-cell fate mapping. Our methodology will greatly facilitate the understanding of morphogenetic processes underlying mammalian organogenesis.

developmental biology↗

Epigenetic priming of embryonic lineages in the mammalian epiblast

Understanding the diversification of mammalian cell lineages is an essential to embryonic development, organ regeneration and tissue engineering. Shortly after implantation in the uterus, the pluripotent cells of the mammalian epiblast generate the three germ layers: ectoderm, mesoderm and endoderm1. Although clonal analyses suggest early specification of epiblast cells towards particular cell lineages2-4, single-cell transcriptomes do not identify lineage-specific markers in the epiblast5-11 and thus, the molecular regulation of such specification remains unknow. Here, we studied the epigenetic landscape of single epiblast cells, which revealed lineage priming towards endoderm, ectoderm or mesoderm. Unexpectedly, epiblast cells with mesodermal priming show a strong signature for the endothelial/endocardial fate, suggesting early specification of this lineage aside from other mesoderm. Through clonal analysis and live imaging, we show that endothelial precursors show early lineage divergence from the rest of mesodermal derivatives. In particular, cardiomyocytes and endocardial cells show limited lineage relationship, despite being temporally and spatially co-recruited during gastrulation. Furthermore, analysing the live tracks of single cells through unsupervised classification of cell migratory activity, we found early behavioral divergence of endothelial precursors shortly after the onset of mesoderm migration towards the cardiogenic area. These results provide a new model for the phenotypically silent specification of mammalian cell lineages in pluripotent cells of the epiblast and modify current knowledge on the sequence and timing of cardiovascular lineages diversification12,13.

developmental biology↗