Abstract
The sea cucumber (Apostichopus japonicus) has become a good model organism for studying environmentally-induced aestivation by a marine invertebrate more recently. In the present study, we hypothesized that miRNA-200-3p may contribute to establish rapid biological control to regulate fatty acid metabolism during a estivation. The peroxisomal bi-functional enzyme (EHHADH) is a crucial participant of the classical peroxisomal fatty acid β-oxidation pathway, the relative mRNA transcripts and protein expressions of EHHADH were analyzed in intestine from sea cucumbers experienced long-term aestivation. Both mRNA transcripts and protein expressions of EHHADH in intestine decreased significantly during deep-aestivation as compared with non-aestivation controls. Analysis of the 3′ UTR of AjEHHADH showed the presence of a conserved binding site for miR-200-3p. Level of miR-200-3p showed an inverse correlation with EHHADH mRNA transcripts and protein levels in intestine, implicating miR-200-3p may directly targeted AjEHHADH by inducing the degradation of AjEHHADH mRNA in the aestivating sea cucumber, further dual-luciferase reporter assay validated the predicted role of miRNA-200-3p in regulating AjEHHADH. In order to further understand their regulatory mechanism, we conducted the functional experiment in vivo. The overexpression of miR-200-3p in sea cucumber significantly decreased mRNA and protein expression levels of AjEHHADH. Taken together, these findings suggested the potential contribution of miRNA-200-3p to the fatty acid metabolism by regulating AjEHHADH during aestivation in sea cucumber.
Highlights
The sea cucumber, Apostichopus japonicus, is an important species farmed in the Chinese aquaculture industry and recently has been embraced as a model organism for studying aestivation in marine invertebrates (Chen et al, 2013; Chen & Storey, 2014; Chen, Zhu & Storey, 2016; Chen et al, 2016)
simple modular architecture research tool (SMART) analysis indicated that the sea cucumber AjEHHADH protein contained two enoyl-CoA hydratase/isomerase (ECH) domains and a 3-hydroxyacyl-CoA dehydrogenase NAD binding domain (Fig. 1) (Gene accession No KY328747KY)
Results showed that the 3 untranslated regions (UTRs) of sea cucumber AjEHHADH contained binding sequences for miR-200-3p starting from residue 2,139 within the 3 UTR sequence
Summary
The sea cucumber, Apostichopus japonicus, is an important species farmed in the Chinese aquaculture industry and recently has been embraced as a model organism for studying aestivation in marine invertebrates (Chen et al, 2013; Chen & Storey, 2014; Chen, Zhu & Storey, 2016; Chen et al, 2016) This animal becomes inactive when water temperature rises over 18 ◦C and is induced to aestivation when temperatures rise over 25 ◦C during the. The optimal regulatory mechanisms for aestivation survival are those that can broadly control multiple metabolic processes, can be coordinated by extracellular stimuli, are induced and readily reversed Among these mechanisms, post-transcription regulation of various transcripts by microRNAs (miRNAs) has become a topic of interest for many researchers
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