Modular, Cascade-like Transcriptional Program of Regeneration in Stentor

The giant ciliate Stentor coeruleus is a classical model system for studying regeneration and morphogenesis at the level of a single cell. The anterior of the cell is marked by an array of cilia, known as the oral apparatus, which can be induced to shed and regenerate in a series of reproducible morphological steps, previously shown to require transcription. If a cell is cut in half, each half will regenerate an intact cell, including a new oral apparatus in the posterior half. We used RNAseq to assay the dynamic changes in Stentor’s transcriptome during regeneration, after both oral apparatus shedding and bisection, allowing us to identify distinct temporal waves of gene expression. By comparing transcriptional profiles of different regeneration events in the same species, we were able to classify regeneration genes into groups based on their potential role for regeneration in distinct cell regeneration paradigms, and provides new insight into how a single cell can coordinate complex morphogenetic pathways to regenerate missing structures. RNA-seq time series of regeneration paradigms in Stentor: 1. sucrose-shocked-induced OA regeneration (10 timepoints, 4 replicates) 2. sucrose-shocked-induced OA regeneration in the presence of cycloheximide ( 5 timepoints, 3 replicates) or in the presence of DMSO (5 timepoints, 3 replicates) 3. bisected cells (anterior and posterior fragments both assayed -- 6 timepoints 3 replicates)