Tissue homeostasis in sponges: quantitative analysis of cell proliferation and apoptosis. Halisarca dujardinii

Published: 11 February 2022| Version 1 | DOI: 10.17632/hhmc8t6gv7.1
Contributors:
Nikolay Melnikov, Andrey Lavrov

Description

Here we provide data underlying the upcoming publication "Tissue homeostasis in sponges: quantitative analysis of cell proliferation and apoptosis". These data provide a quantitative description of cell proliferation and apoptosis in intact tissues of marine sponge, Halisarca dujardinii (class Demospongiae). Tissues of Halisarca are highly proliferative, indicating either high rates of cell turnover or body growth. The main fraction of proliferating cells are choanocytes, food-entrapping flagellated cells of the aquiferous system. They also seem to be the main source of new mesohyl cells. The numbers of apoptotic cells in intact tissues are insignificant; both choanocytes and mesohyl cells undergo apoptosis. CLSM folder contains Z-stacks obtained via confocal laser scanning microscope. Z-stacks were processed in Bitplane Imaris software and saved in .ems format. Apoptosis subfolder contains Z-stacks obtained during the experiment "Investigation of apoptotic activity: staining of intact tissues". Tissues are stained with DAPI (channel 1; cell nuclei), CellEvent (channel 2; nuclei of apoptotic cells), anti-acetylated alpha-tubulin antibodies (channel 3; choanocyte flagella). Intact tissues subfolder contains Z-stacks obtained during the experiment "Study of cell proliferation with CLSM". Tissues are stained with DAPI (channel 1; cell nuclei), EdU (channel 3; 12 hours of incubation; nuclei of S-phase cells), anti-phospho-histone 3 antibodies (channel 4; nuclei of M-phase cells), anti-acetylated alpha-tubulin antibodies (channel 2; choanocyte flagella). Pulse-chase subfolder contains Z-stacks obtained during the experiment "EdU tracking in Halisarca dujardinii". Tissues are stained with DAPI (channel 1; cell nuclei) and EdU (channel 2; 12 hours of incubation; nuclei of S-phase cells). In vivo apoptosis subfolder contains Z-stacks obtained during the experiment "Investigation of apoptotic activity in intact tissues". Cell suspension is stained with Hoechst (channel 1; cell nuclei), CellEvent (channel 2; nuclei of apoptotic cells), TMRE (channel 3; functional mitochondria). Flow cytometry folder contains raw .fcs files. Cell cycle subfolder contains files obtained during the experiment "Evaluation of cell cycle distribution". Cell suspensions are stained with propidium iodide (PE-A channel). In vivo apoptosis subfolder contains files obtained during the experiment "Investigation of apoptotic activity in intact tissues". Cell suspensions are stained with CellEvent (FITC-A channel) and TMRE (PE-A channel). Apoptosis fixed contains files obtained during the experiment "Investigation of apoptotic activity in intact tissues". Cell suspensions are stained with CellEvent (FITC-A channel)

Files

Steps to reproduce

Sampling and all used protocols are described in the paper.

Categories

Zoology, Cell Biology, Confocal Microscopy, Evolutionary Developmental Biology, Flow Cytometry

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