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Quantitative proteomics reveals remodeling of protein repertoire across life phases of Daphnia pulex

ABSTRACT: Microcrustacean Daphnia is becoming a model organism of choice for ecological and developmental genomics. Yet, Daphnia proteomics resources have so far been rudimental. This study presents LC-MS2/SPS-MS3 proteomes of adult females, juveniles, asexually produces embryos and sexually produced diapausing embryos capable of surviving freezing and desiccation. Samples were fractionated using high pH reversed-phase chromatography. 12 farctions were analyzed by LC-MS2/SPS-MS3. The samples are labeled as follows: E1 (128n), E2 (128c), E3 (129n); 3 biological replicates of asexually produced embryos J1 (127n), J2 (127c), J3 (131n); 3 biological replicates of juveniles M1 (129c), M2 (130n), M3 (130c); 3 biological replicates of adult females Eph (126); a single ephippium sample (sexually produced diapausing embryos).

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Daphnia Pulex (ncbitaxon:6669)

SUBMITTER: Lev Yampolski

PROVIDER: MSV000083711 | MassIVE | Mon Apr 22 10:19:00 BST 2019

REPOSITORIES: MassIVE

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Project description:The marine copepod Calanus finmarchicus is the most abundant zooplankton species in the northern regions of the Atlantic Ocean and the Barents Sea. Very little is known about the molecular mechanisms underlying critical processes associated with this species’ complex life history (e.g., ontogenetic development, reproduction, molting, diapause) and physiology (e.g., digestion, neural processes, and membrane physiology). This study analyzed patterns of gene expression of C. finmarchicus samples collected from the Gulf of Maine (Northwest Atlantic Ocean) using a 1,000 expressed sequence tag (EST) microarray designed to assay genes of known physiological function and hypothesized ecological importance for C. finmarchicus. Replicate analyses compared adult females and final-stage juveniles (Copepodite-5) collected from surface (0-30m) and deep (130-170m) layers. Environmental data include detailed characterization of biological, chemical, and physical oceanographic parameters in the sampled water packets. All data were screened for artifacts, normalized and selected using a fold-change filter prior to analysis. Replicate comparisons were analyzed by Significance Analysis of Microarrays (SAM; Stanford University Labs) with a control for False Discovery Rate (FDR) and with Principle Component Analysis with evaluation of significance by one- or two-sample t-test in Acuity Microarray Informatics Software (Molecular Devices, Inc.). Gene Ontology Enrichment Analysis was carried out using GOEAST (http://omicslab.genetics.ac.cn/GOEAST/index.php) to assess functional relationships of selected genes and/or proteins. The results indicated: up-regulation of genes involved in cell division, protein synthesis and mating in deep females and juveniles; up-regulation of genes related to cellular homeostasis, circadian behavior and nervous system development in surface females; and up-regulation of genes related to muscle development and protein catabolism in deep juveniles versus deep females. KEGG pathway analysis using the Blast2GO suite (http://www.blast2go.org/) indicated: up-regulation of genes encoding enzymes related to the citrate cycle and anaerobic metabolism in deep females and juveniles; and up-regulation of genes encoding enzymes related to energy metabolism and osmoregulation in surface females.

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Quantitative proteomics reveals remodeling of protein repertoire across life phases of Daphnia pulex

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