Transcriptome Analysis of Larval Segment Formation and Secondary Loss in the Echiuran Worm Urechis unicinctus.

Xitan Hou,Tingting Zhang,Zhifeng Zhang,Di Zhou,Qi Li,Zhenkui Qin,Dexu Kong,Yueyang Xie,Maokai Wei

doi:10.3390/ijms20081806

Xitan Hou, Tingting Zhang + Show 7 more

Open Access

https://doi.org/10.3390/ijms20081806

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Abstract

The larval segment formation and secondary loss in echiurans is a special phenomenon, which is considered to be one of the important characteristics in the evolutionary relationship between the Echiura and Annelida. To better understand the molecular mechanism of this phenomenon, we revealed the larval transcriptome profile of the echiuran worm Urechis unicinctus using RNA-Seq technology. Twelve cDNA libraries of U. unicinctus larvae, late-trochophore (LT), early-segmentation larva (ES), segmentation larva (SL), and worm-shaped larva (WL) were constructed. Totally 243,381 unigenes were assembled with an average length of 1125 bp and N50 of 1836 bp, and 149,488 unigenes (61.42%) were annotated. We obtained 70,517 differentially expressed genes (DEGs) by pairwise comparison of the larval transcriptome data at different developmental stages and clustered them into 20 gene expression profiles using STEM software. Based on the typical profiles during the larval segment formation and secondary loss, eight signaling pathways were enriched, and five of which, mTOR, PI3K-AKT, TGF-β, MAPK, and Dorso-ventral axis formation signaling pathway, were proposed for the first time to be involved in the segment formation. Furthermore, we identified 119 unigenes related to the segment formation of annelids, arthropods, and chordates, in which 101 genes were identified in Drosophila and annelids. The function of most segment polarity gene homologs (hedgehog, wingless, engrailed, etc.) was conserved in echiurans, annelids, and arthropods based on their expression profiles, while the gap and pair-rule gene homologs were not. Finally, we verified that strong positive signals of Hedgehog were indeed located on the boundary of larval segments using immunofluorescence. Data in this study provide molecular evidence for the understanding of larval segment development in echiurans and may serve as a blueprint for segmented ancestors in future research.

Highlights

Echiurans are a group of marine benthic invertebrates including approximately 230 species, which all inhabit marine environments from the intertidal zone to thousands of meters in the deep sea [1,2,3], such as Bonellia viridis in coastal sediment and Hydroides elegans in the deep sea
NK genes may be involved in the formation and maintenance of larval segments in U. unicinctus
Our study presented the first transcriptome analysis focusing on the gene expression profiles of the segment formation and secondary loss in U. unicinctus, in which 101 genes were identified involving in the segment formation as in Drosophila and annelids

Summary

Introduction

Echiurans are a group of marine benthic invertebrates including approximately 230 species, which all inhabit marine environments from the intertidal zone to thousands of meters in the deep sea [1,2,3], such as Bonellia viridis in coastal sediment and Hydroides elegans in the deep sea. It has been a controversial issue whether the echiurans belong to Annelida or a separate phylum Echiura [4]. Based on the obvious difference between the larval segmentation in echiurans and the typical segmentation in annelids, it is of great significance to explore the molecular mechanism of larval segment formation in echiurans

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