bioRxiv Science⌕ Search

bioRxiv · 10.1101/2024.03.08.584197

Exploration of phosphoproteomic association during epimorphic regeneration

Abstract

Unravelling the intricate patterns of site-specific protein phosphorylation during Epimorphic regeneration holds the key to unlocking the secrets of tissue complexity. Understanding these precise modifications and their impact on protein function could shed light on the remarkable regenerative capacity of tissues, with potential implications for therapeutic interventions. In this study we have systematically mapped the global phosphorylation modifications within regenerating tissue of zebrafish caudal fins, elucidating the intricate landscape of signalling pathway associate with the regeneration process. A total of 74 and 440 proteins were found undergoing differentially phosphorylated during the process of regeneration from 12hpa to 7dpa against control based on TiO2 column enrichment and immuno precipitation using phosphoserine, phosphothreonine and phosphotyrosine antibodies respectively. Interestingly 95% of the proteins identified from TiO2 enrichment method were also found to be identified through the phosphoprotein antibody pull down method impacting the high accuracy and significance of the methods and greater association of the 70 proteins undergoing differential phosphorylation during the process of regeneration. Whole mount immunohistochemistry analysis reveals high association of phosphorylation at 1dpa, 2dpa and 3 dpa regeneration time points. Based on network pathway analysis it was evident that Fc Receptor-mediated Phagocytosis in Macrophages and Monocytes, Actin cytoskeleton signaling, HGF signaling and Insulin receptor signaling are the most highly associated network pathways for regeneration through differential phosphorylation. This research enhances our comprehension on protein post-translational modification in the context of zebrafish caudal fin tissue regeneration, shedding light on its prospective application in the field of regenerative medicine.

Source connections

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Banu, S., V, A. P., Mandal, K., Idris, M. M.. 2024-03-13. Exploration of phosphoproteomic association during epimorphic regeneration. https://doi.org/10.1101/2024.03.08.584197

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

KEEP EXPLORING

Related preprints

Hyaluronic acid dynamics are regulated by neural crest cells and NEDD4 in the developing cardiac outflow tract valves

The extracellular matrix (ECM) is essential for cardiac valve development, with hyaluronic acid (HA) playing a critical role in cardiac cushion formation. While HA is required for atrioventricular valve development, its role in semilunar valve formation remains poorly understood. The ubiquitin ligase NEDD4 is essential for cardiac outflow tract development, particularly within cardiac neural crest cells, however its role in semilunar valve development has been unexplored. Here, we demonstrate that neural crest-specific deletion of NEDD4 in mice disrupts outflow tract valve development, resulting in abnormal leaflet number and morphology. Analysis of valve ECM revealed a specific reduction in HA within the neural crest-derived valve mesenchyme, while other ECM components such as fibronectin and versican remained unaffected. In contrast, deletion of NEDD4 in endocardium-derived valve mesenchyme did not alter HA abundance. Using NEDD4-deficient neural crest cells, we further demonstrate a cell-autonomous requirement for NEDD4 in regulating HA abundance, with in vitro evidence supporting impaired HA synthesis rather than increased turnover of HA in the ECM. Our findings identify a previously unrecognised role for NEDD4 in regulating HA synthesis in cardiac neural crest cells, revealing a cell-type-specific mechanism governing semilunar valve development and highlighting distinct cellular requirements for ECM regulation across cardiac valve populations.

developmental biology↗

Endogenous Mechanical Reporters Encode the Global Mechanical Landscape of Morphogenesis

Developmental systems usually comprise multiple genetically programmed morphogenetic events within a mechanically continuous environment, yet how individual processes interact and contribute to the overall mechanical landscape remains unknown. Here, we introduce the concept of mechanical reporter: endogenous tissues function as in situ probes of the embedded mechanical landscape. We exemplified cephalic furrow (CF) migration as a mechanical reporter in gastrulating Drosophila. Using optogenetics and genetics, we revealed that CF displacement reflects the combined effect of both force and tissue mechanical properties. By incorporating condition-specific mechanical landscape into a continuum minimal-response model, we developed a framework to quantitatively predict the reporter dynamics. Using different genetic mutants, we successfully decoded changes in the mechanical landscape from CF dynamics in gastrulating Drosophila. Overall, we established a predictive framework that uses CF dynamics to infer changes in the global mechanical landscape. Beyond this, the concept of mechanical reporter provides a general strategy for systematically understanding and decoding complex mechanical environments in living systems.

developmental biology↗

Dissecting human embryonic patterning during organogenesis reveals a rostral midline progenitor population for mesoendoderm induction

Understanding how human organogenesis is accomplished via intricate spatiotemporal patterning of organ primordia requires spatial atlases at cellular resolution. However, existing human embryo spatial atlases are built using sequencing-based platforms such as Stereo-seq, which have limited spatial resolution and blurred anatomical boundaries, leading to misclassification across adjacent cells. Here, we integrate in situ imaging-based transcriptomics (Xenium) with transcriptome-wide spatial profiling (Visium-HD) of Carnegie stages 9-12 embryos to delineate the patterning of core embryonic tissues at single-cell resolution, including the brain, spinal cord, somites, neural crest, notochord, heart, and gut tube. A new finding enabled by spatial single-cell resolution is the identification of rostral midline progenitors (RMPs), a cell population co-expressing SOX2, TBXT, and OTX2. Transcriptionally, RMPs are distinct from both neuromesodermal progenitors (NMPs) and prechordal mesoderm, and in vivo clonal barcoding in mice indicates that RMPs share a common developmental origin with rostral neural ectoderm. To interrogate their functional significance, we leveraged our spatial transcriptome data to recapitulate RMP-like cells from human pluripotent stem cells and showed that these cells can induce epiblast-like cells to acquire mesendodermal and endodermal fates, while themselves adopting an ectodermal fate consistent with their in vivo counterparts. This work provides a 3D, stage-resolved molecular cell atlas of tissue patterning in human embryo that serves as a reference for in vitro models.

developmental biology↗