Identification, Characterization, and Diel Pattern of Expression of Canonical Clock Genes in Nephrops norvegicus (Crustacea: Decapoda) Eyestalk

Type Article
Date 2015-11
Language English
Author(s) Sbragaglia Valerio1, Lamanna FrancescoORCID2, Mat Audrey3, Rotllant GuiomarORCID1, Joly Silvia1, Ketmaier Valerio4, de La Iglesia Horacio O.3, Aguzzi Jacopo1
Affiliation(s) 1 : ICM CSIC, Inst Marine Sci, Barcelona, Spain.
2 : Univ Potsdam, Inst Biochem & Biol, Unit Evolutionary Biol Systemat Zool, Potsdam, Germany.
3 : Univ Washington, Dept Biol, Seattle, WA 98195 USA.
4 : Univ Roma La Sapienza, Dept Biol & Biotechnol Charles Darwin, I-00185 Rome, Italy.
Source Plos One (1932-6203) (Public Library Science), 2015-11 , Vol. 10 , N. 11 , P. e0141893. (17p.)
DOI 10.1371/journal.pone.0141893
WOS© Times Cited 24
Abstract

The Norway lobster, Nephrops norvegicus, is a burrowing decapod with a rhythmic burrow emergence (24 h) governed by the circadian system. It is an important resource for European fisheries and its behavior deeply affects its availability. The current knowledge of Nephrops circadian biology is phenomenological as it is currently the case for almost all crustaceans. In attempt to elucidate the putative molecular mechanisms underlying circadian gene regulation in Nephrops, we used a transcriptomics approach on cDNA extracted from the eyestalk, a structure playing a crucial role in controlling behavior of decapods. We studied 14 male lobsters under 12-12 light-darkness blue light cycle. We used the Hiseq 2000 Illumina platform to sequence two eyestalk libraries (under light and darkness conditions) obtaining about 90 millions 100-bp paired-end reads. Trinity was used for the de novo reconstruction of transcriptomes; the size at which half of all assembled bases reside in contigs (N50) was equal to 1796 (light) and 2055 (darkness). We found a list of candidate clock genes and focused our attention on canonical ones: timeless, period, clock and bmal1. The cloning of assembled fragments validated Trinity outputs. The putative Nephrops clock genes showed high levels of identity (blastx on NCBI) with known crustacean clock gene homologs such as Eurydice pulchra (period: 47%, timeless: 59%, bmal1: 79%) and Macrobrachium rosenbergii (clock: 100%). We also found a vertebrate-like cryptochrome 2. RT-qPCR showed that only timeless had a robust diel pattern of expression. Our data are in accordance with the current knowledge of the crustacean circadian clock, reinforcing the idea that the molecular clockwork of this group shows some differences with the established model in Drosophila melanogaster.

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Sbragaglia Valerio, Lamanna Francesco, Mat Audrey, Rotllant Guiomar, Joly Silvia, Ketmaier Valerio, de La Iglesia Horacio O., Aguzzi Jacopo (2015). Identification, Characterization, and Diel Pattern of Expression of Canonical Clock Genes in Nephrops norvegicus (Crustacea: Decapoda) Eyestalk. Plos One, 10(11), e0141893. (17p.). Publisher's official version : https://doi.org/10.1371/journal.pone.0141893 , Open Access version : https://archimer.ifremer.fr/doc/00483/59476/