Transcriptional Profiling of Predator-Induced Phenotypic Plasticity in Daphnia Pulex Andrey Rozenberg1†, Mrutyunjaya Parida2†, Florian Leese1,4, Linda C

Transcriptional Profiling of Predator-Induced Phenotypic Plasticity in Daphnia Pulex Andrey Rozenberg1†, Mrutyunjaya Parida2†, Florian Leese1,4, Linda C

Rozenberg et al. Frontiers in Zoology (2015) 12:18 DOI 10.1186/s12983-015-0109-x RESEARCH Open Access Transcriptional profiling of predator-induced phenotypic plasticity in Daphnia pulex Andrey Rozenberg1†, Mrutyunjaya Parida2†, Florian Leese1,4, Linda C. Weiss1,3, Ralph Tollrian1* and J. Robert Manak2 Abstract Background: Predator-induced defences are a prominent example of phenotypic plasticity found from single-celled organisms to vertebrates. The water flea Daphnia pulex is a very convenient ecological genomic model for studying predator-induced defences as it exhibits substantial morphological changes under predation risk. Most importantly, however, genetically identical clones can be transcriptionally profiled under both control and predation risk conditions and be compared due to the availability of the sequenced reference genome. Earlier gene expression analyses of candidate genes as well as a tiled genomic microarray expression experiment have provided insights into some genes involved in predator-induced phenotypic plasticity. Here we performed the first RNA-Seq analysis to identify genes that were differentially expressed in defended vs. undefended D. pulex specimens in order to explore the genetic mechanisms underlying predator-induced defences at a qualitatively novel level. Results: We report 230 differentially expressed genes (158 up- and 72 down-regulated) identified in at least two of three different assembly approaches. Several of the differentially regulated genes belong to families of paralogous genes. The most prominent classes amongst the up-regulated genes include cuticle genes, zinc-metalloproteinases and vitellogenin genes. Furthermore, several genes from this group code for proteins recruited in chromatin- reorganization or regulation of the cell cycle (cyclins). Down-regulated gene classes include C-type lectins, proteins involved in lipogenesis, and other families, some of which encode proteins with no known molecular function. Conclusions: The RNA-Seq transcriptome data presented in this study provide important insights into gene regulatory patterns underlying predator-induced defences. In particular, we characterized different effector genes and gene families found to be regulated in Daphnia in response to the presence of an invertebrate predator. These effector genes are mostly in agreement with expectations based on observed phenotypic changes including morphological alterations, i.e., expression of proteins involved in formation of protective structures and in cuticle strengthening, as well as proteins required for resource re-allocation. Our findings identify key genetic pathways associated with anti-predator defences. Keywords: Daphnia; Inducible defences; Phenotypic plasticity; Predator-prey interaction; RNA-Seq; Transcriptomics; Morphology; Gene-environment interactions Background [4] make the system highly attractive for evolutionary Thecommonfreshwatermicro-crustaceanDaphnia has ecology research and provides the unique opportunity to become a model organism for many biological disciplines study ecological traits with the aid of emerging molec- [1–6]. The extensive knowledge of its ecology [5, 7, 8] ular biological tools. One of the most intriguing eco- and its biological responses to environmental changes logical responses of Daphnia species to environmental [3, 9, 10] together with the availability of genomic resources changes is their ability to develop different phenotypes given the same genetic background, a phenomenon called *Correspondence: [email protected] phenotypic plasticity. Prominent examples of phenotypic †Equal contributor 1Department of Animal Ecology, Evolution and Biodiversity, Ruhr University plasticity include inducible defences. Bochum, Universitaetsstrasse 150, 44801 Bochum, Germany Inducible defences are interpreted as adaptations to Full list of author information is available at the end of the article heterogeneous predation risks and are found in many © 2015 Rozenberg et al. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Rozenberg et al. Frontiers in Zoology (2015) 12:18 Page 2 of 13 organisms from protists to vertebrates [11–13]. Daphnia D. pulex this habit is displayed only by some clones evolved sensitivity against specific chemical compounds, [34–36] (LCW, unpublished observations). which are unintentionally emitted by their predators. Instability of environmental conditions (periodicity of These so-called kairomones serve as signals which prompt predation risk, different predators) and costs of defences the daphnid prey to develop individuals which are better explain the inducible nature of the defensive morphs. defended. Previous work has shown that different preda- This is also in line with the fact that the neck-teeth are tors, e.g. fish and the phantom midge Chaoborus spp., can present only in certain juvenile instars when the daphnids induce different, sometimes opposite phenotypic reac- reach preferred prey size of their gape-limited predators tions in the same species or clone [10, 14–17]. This means [10, 13, 29, 37, 38]. that the genome must encode multiple developmental Based on the experimental evidence we can make programs triggered by environmental conditions. Induced the following predictions regarding the underlying func- defences in Daphnia include prominent morphological tional classes of effector genes that might contribute to modifications: from tiny cuticular teeth to very elongated Daphnia’s anti-predatory response (Fig. 1): tail and head spines, helmets or even giant crests [18–21], but also changes in life history and different behaviours, 1. the structural changes in the cuticle are expected to which ultimately all act as deterrents to encounter, capture mirror changes in the amounts/types of and ingestion by the predator [9, 22–25]. cuticle-associated proteins; In the model species Daphnia pulex, kairomones from 2. life history modifications are expected to be the phantom-midge larvae Chaoborus trigger produc- controlled by several physiological changes affecting tion of neck-teeth, the most easily detectable trait, and both somatic growth and reproduction; overall hardening of the cuticle [26]. These external, 3. one can predict that other metabolic functions cuticle-associated alterations effectively protect juveniles should be down regulated under predation risk in from predation [27, 28]. At the same time, induced order to allocate energy resources primarily to the D. pulex females shift resources from reproduction to above mentioned pathways; somatic growth, thereby reaching maturity at a larger size 4. all levels of the response must ultimately be and producing less but larger offspring [29–31]. Vertical controlled by cascades of receptors, humoral factors, migration is deemed to comprise the main behavioural signalling pathways and transcription factors. reaction to the presence of the predator in D. pulex:thus, Chaoborus-induced specimens prefer shallower depths in Currently, the technique of choice suitable for address- comparison to control specimens [32, 33]. Chaoborus is ing response patterns in gene expression at a genome- an ambush predator, such that Daphnia is also expected wide level with potentially unlimited depth of coverage is to reduce its swimming speed, although in the case of RNA-Seq [39–41]. The availability of the D. pulex draft Fig. 1 Presumptive scheme of physiological and morphological changes during kairomone-induction in D. pulex Rozenberg et al. Frontiers in Zoology (2015) 12:18 Page 3 of 13 genome [4] greatly facilitates the power of such analy- h. Subsequently, specimens were stored at -20 °C until ses in that RNA-Seq reads can be specifically mapped RNA extraction. Three independent induction experi- to a particular genomic location. Investigations of the ments were performed leading to 90 juveniles in each genome-environment interactions in Daphnia are ongo- group (induction and control) in total, which were pooled ing, with the results of the first analyses of differential gene together to level individual variation. expression patterns in ecological experiments recently becoming available [42–45]. A number of features have RNA extraction and sequencing been discovered that point to an ecological responsive- Total RNA was extracted with the TRIzol Reagent (Life ness of the D. pulex genome; e.g. a large overall number of Technologies, Carlsbad, CA, USA) according to the man- genes, organized in the many families of paralogous genes ufacturer’s protocol with modifications. DNA was fur- that in many cases do not show homologies to genes in ther depleted with DNase treatment (TurboDNase, Life other organisms, but show differential expression under Technologies, Carlsbad, CA, USA) and its absence was different environmental conditions [4, 46–49]. confirmed afterwards via PCR with primers spanning While preliminary analysis of the transcriptomic exon-intron boundaries in an α-tubulin gene. Quality and responses of D. pulex to the predator was performed ear- amount of the purified RNA in the samples was ana- lier [4], gene expression was assessed with tiling microar- lyzed on the Experion System

View Full Text

Details

  • File Type
    pdf
  • Upload Time
    -
  • Content Languages
    English
  • Upload User
    Anonymous/Not logged-in
  • File Pages
    13 Page
  • File Size
    -

Download

Channel Download Status
Express Download Enable

Copyright

We respect the copyrights and intellectual property rights of all users. All uploaded documents are either original works of the uploader or authorized works of the rightful owners.

  • Not to be reproduced or distributed without explicit permission.
  • Not used for commercial purposes outside of approved use cases.
  • Not used to infringe on the rights of the original creators.
  • If you believe any content infringes your copyright, please contact us immediately.

Support

For help with questions, suggestions, or problems, please contact us