Quantitative profiling of Drosophila melanogaster Dscam1 isoforms reveals no changes in splicing after bacterial exposure

PLoS One. 2014 Oct 13;9(10):e108660. doi: 10.1371/journal.pone.0108660. eCollection 2014.

Abstract

The hypervariable Dscam1 (Down syndrome cell adhesion molecule 1) gene can produce thousands of different ectodomain isoforms via mutually exclusive alternative splicing. Dscam1 appears to be involved in the immune response of some insects and crustaceans. It has been proposed that the diverse isoforms may be involved in the recognition of, or the defence against, diverse parasite epitopes, although evidence to support this is sparse. A prediction that can be generated from this hypothesis is that the gene expression of specific exons and/or isoforms is influenced by exposure to an immune elicitor. To test this hypothesis, we for the first time, use a long read RNA sequencing method to directly investigate the Dscam1 splicing pattern after exposing adult Drosophila melanogaster and a S2 cell line to live Escherichia coli. After bacterial exposure both models showed increased expression of immune-related genes, indicating that the immune system had been activated. However there were no changes in total Dscam1 mRNA expression. RNA sequencing further showed that there were no significant changes in individual exon expression and no changes in isoform splicing patterns in response to bacterial exposure. Therefore our studies do not support a change of D. melanogaster Dscam1 isoform diversity in response to live E. coli. Nevertheless, in future this approach could be used to identify potentially immune-related Dscam1 splicing regulation in other host species or in response to other pathogens.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alternative Splicing*
  • Animals
  • Cell Adhesion Molecules
  • Drosophila Proteins / genetics
  • Drosophila Proteins / metabolism*
  • Drosophila melanogaster
  • Escherichia coli / genetics
  • Neural Cell Adhesion Molecules / genetics
  • Neural Cell Adhesion Molecules / metabolism*
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism*
  • Sequence Analysis, RNA

Substances

  • Cell Adhesion Molecules
  • Drosophila Proteins
  • Dscam1 protein, Drosophila
  • Neural Cell Adhesion Molecules
  • Protein Isoforms

Grants and funding

Support was provided to DS by Fonds Wetenschappelijk Onderzoek - Vlaanderen (FWO), (http://www.fwo.be/en/) (G059611N) BELSPO IUAP VII-20 “WIBRAIN project”, to SAOA by Volkswagen Foundation Postdoctoral Fellowship (http://www.volkswagenstiftung.de/) Grant numbers: 83516 and 86020; German Research Foundation (DFG) (http://www.dfg.de/en/) Grant number: AR 872/1-1 within SPP 1399, to WC by Berlin Institute for Medical Systems Biology of the Federal Ministry for Education and Research (BMBF) and Senate of Berlin (www.bmbf.de, www.berlin.de/sen/wtf) Grant number: 0315362, and to WS by the China Scholarship Council (CSC) (http://en.csc.edu.cn/) Grant number: 2010623030. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.