RNA-seq dependent transcriptional analysis unveils gene expression profile in the intestine of sea cucumber Apostichopus japonicus during aestivation

Abstract

The seasonal marine, the sea cucumber Apostichopus japonicus (Selenka, 1867), cycles annually between periods of torpor when water temperature is above about 25°C in summer and active life when temperature is below about 18°C. This species is a good candidate model organism for studies of environmentally-induced aestivation in marine invertebrates. Previous studies have examined various aspects of aestivation of A. japonicus, however, knowledge of the molecular regulation underpinning these events is still fragmentary. In the present study, we constructed a global gene expression profile of the intestine tissue of A. japonicus using RNA-seq to identify transcriptional responses associated with transitions between different states: non-aestivation (NA), deep-aestivation (DA), and arousal from aestivation (AA). The analysis identified 1245 differentially expressed genes (DEGs) between DA vs. NA states, 1338 DEGs between AA vs. DA, and 1321 DEGs between AA vs. NA using the criteria |Log2Ratio|≥1 and FDR≤0.001. Of these, 25 of the most significant DEGs were verified by real-time PCR, showing trends in expression patterns that were almost in full concordance between the two techniques. GO analysis revealed that for DA vs. NA, 24 metabolic associated processes were highly enriched (corrected p value<0.05) whereas for AA vs. NA, 12 transport and metabolism associated processes were significantly enriched (corrected p value<0.05). Pathways associated with aestivation were also mined, and indicated that most DEGs were enriched in metabolic and signal transduction pathways in the deep aestivation stage. Two up pathways were significantly enriched at the arousal stage (ribosome and metabolism of xenobiotics by cytochrome P450 pathway). A set of key DEGs was identified that may play vital roles in aestivation; these involved metabolism, detoxification and tissue protection, and energy-expensive processes. Our work presents an overview of dynamic gene expression in torpor-arousal cycles during aestivation of A. japonicus and identifies a series of candidate genes and pathways for further research on the molecular mechanisms of aestivation.

Keywords: Aestivation–arousal cycles; Differential gene expression; Echinoderm; Environmental stress; Hypometabolism.