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Rusciano, M. L.

Publications and source records attributed to Rusciano, M. L..

3 recordsLinked to original sources

Single Nucleus Profiling Highlights the All-Brain Echinoderm Nervous System

Metazoans comprise diverse tissues and cell types, each essential for the survival of the organism. Most of these cell types are established early in embryogenesis and persist into adulthood. However, in indirectly developing sea urchins, the continuity between embryonic and adult stages is thus interrupted by a planktonic larval stage that undergoes complete metamorphosis. In addition, while the gene regulatory networks governing distinct embryonic and larval cell lineages are well studied, the molecular and morphological identities of post-metamorphic sea urchin cell types remain poorly understood. Here, we reconstructed the cell type atlas of post-metamorphic Paracentrotus lividus juveniles using single-nucleus transcriptomics, shedding light on the conservation of genetic regulatory mechanisms in post-metamorphic cell types. We identified cell signatures corresponding to eight distinct cell type groups and analyzed at least twenty-nine neuronal cell families, including fifteen unique photoreceptor cell signatures. By combining single cell transcriptomics with spatial gene expression analysis and high-resolution electron microscopy, we identified homologues of vertebrate neuronal genes and photoreceptive opsins expressed throughout the sea urchin body. These findings provide evidence that the echinoderm body plan is not only predominantly head-like, but also exhibits an all-brain organization in an animal previously considered to have a primitive nervous system.

evolutionary biology↗

Deep conservation of cis-regulatory elements and chromatin organization in echinoderms uncover ancestral regulatory features of animal genomes

Despite the growing abundance of sequenced animal genomes, we only have detailed knowledge of regulatory organization for a handful of lineages, particularly flies and vertebrates. These two groups of taxa show contrasting trends in the molecular mechanisms of 3D chromatin organization and long-term evolutionary dynamics of cis-regulatory element (CREs) conservation. To help us identify shared versus derived features that could be responsible for the evolution of these different regulatory architectures in animals, we studied the evolution and organization of the regulatory genome of echinoderms, a lineage whose phylogenetic position and relatively slow molecular evolution has proven particularly useful for evolutionary studies. First, using PacBio and HiC data, we generated new reference genome assemblies for two species belonging to two different echinoderm classes: the purple sea urchin Strongylocentrotus purpuratus and the bat sea star Patiria miniata. Second, we characterized their 3D chromatin architecture, identifying TAD-like domains in echinoderms that, like in flies, do not seem to be associated with CTCF motif orientation. Third, we systematically profiled CREs during sea star and sea urchin development using ATAC-seq, comparing their regulatory logic and dynamics over multiple developmental stages. Finally, we investigated sea urchin and sea star CRE evolution across multiple evolutionary distances and timescales, from closely related species to other echinoderm classes and deuterostome lineages. This showed the presence of several thousand elements conserved for hundreds of millions of years, revealing a vertebrate-like pattern of CRE evolution that probably constitutes an ancestral property of the regulatory evolution of animals.

evolutionary biology↗

Long-term live imaging, cell identification and cell tracking in regenerating crustacean legs

High resolution live imaging of regeneration presents unique challenges, due to the nature of the specimens (large mobile animals), the duration of the process (spanning days or weeks), and the fact that cellular resolution must be achieved without damage caused by lengthy exposures to light. Building on previous work that allowed us to image different parts of the process of leg regeneration in the crustacean Parhyale hawaiensis, we present here a method for live imaging that captures the entire process of leg regeneration, spanning up to 10 days, at cellular resolution. Our method includes (1) mounting and long-term live imaging of regenerating legs under conditions that yield high spatial and temporal resolution but minimise photodamage, (2) fixing and in situ staining of the regenerated legs that were imaged, to identify cell fates, and (3) computer-assisted cell tracking to determine the cell lineages and progenitors of identified cells. The method is optimised to limit light exposure while maximising tracking efficiency. Combined with appropriate cell-type-specific markers, this method may allow the description of cell lineages for every regenerated cell type in the limb.

developmental biology↗