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Long-term time-lapse live imaging reveals extensive cell migration during annelid regeneration

Zattara, Eduardo E; Turlington, Kate W; Bely, Alexandra E

Supporting Materials for the article:
Long-term time-lapse live imaging reveals extensive cell migration during annelid regeneration

Eduardo E. Zattara, Kate W. Turlington and Alexandra E. Bely

BMC Developmental Biology, 2016

Includes 11 movies and one compressed file with R code and data tables.

Here we describe a new method for long-duration time-lapse imaging of adults of the small freshwater annelid Pristina leidyi and use this method to investigate its regenerative processes. Specimens are immobilized with tetrodotoxin, resulting in irreversible paralysis yet apparently normal regeneration, and mounted in agarose surrounded by culture water or halocarbon oil, to prevent dehydration but allowing gas exchange. Using this method, worms can be imaged continuously and at high spatial-temporal resolution for up to 5 days, spanning the entire regeneration process. We performed a fine-scale analysis of regeneration growth rate and characterized cell migration dynamics during early regeneration. Our studies reveal the migration of several putative cell types, including one strongly resembling published descriptions of annelid neoblasts, a cell type suggested to be migratory based on "still-shot" studies and long hypothesized to be linked to regenerative success in annelids.
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