Visualizing Calcium Deposition and Cell Proliferation in Juvenile Sea Urchins

To visualize newly deposited CaCO₃ of the skeleton, live animals were incubated with Calcein (Sigma) as described in [41 (link)]. To understand the spatial distribution and quantity of proliferating cells during juvenile sea urchin growth assays of cell proliferation were carried out using the Click-iT® EdU Alexa Flour® 555HCS (Life Technologies) and Click-iT™ EdU Cell Proliferation Kit for Imaging Alexa Flour™ 647 (Thermo Fisher Scientific) following [41 (link)]. Cells were quantified for analyzed images using ImageJ and R. Additional details of Calcein staining, proliferative staining and imaging can be found in Additional file 1: Methods.

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Thompson J.R., Paganos P., Benvenuto G., Arnone M.I, & Oliveri P. (2021). Post-metamorphic skeletal growth in the sea urchin Paracentrotus lividus and implications for body plan evolution. EvoDevo, 12, 3.

Publication 2021

Calcein Click-iT® EdU Alexa Flour® 555HCS Click-iT™ EdU Cell Proliferation Kit for Imaging Alexa Flour™ 647

Animals Assays Caco3 Calcein Cell proliferation Cells Flour Sea urchin Skeleton

Corresponding Organization : Centre for Life

Other organizations : Stazione Zoologica Anton Dohrn

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Variable analysis

independent variables

Incubation of live animals with Calcein (Sigma) to visualize newly deposited CaCO3 of the skeleton
Use of Click-iT® EdU Alexa Flour® 555HCS (Life Technologies) and Click-iT™ EdU Cell Proliferation Kit for Imaging Alexa Flour™ 647 (Thermo Fisher Scientific) to understand the spatial distribution and quantity of proliferating cells during juvenile sea urchin growth

dependent variables

Spatial distribution and quantity of newly deposited CaCO3 of the skeleton
Spatial distribution and quantity of proliferating cells during juvenile sea urchin growth

control variables

Not explicitly mentioned

controls

Positive control: Not specified
Negative control: Not specified

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