Precise staging of beetle horn formation in Trypoxylus dichotomus reveals the pleiotropic roles of doublesex depending on the spatiotemporal developmental contexts

PLoS Genet. 2019 Apr 10;15(4):e1008063. doi: 10.1371/journal.pgen.1008063. eCollection 2019 Apr.

Abstract

Many scarab beetles have sexually dimorphic exaggerated horns that are an evolutionary novelty. Since the shape, number, size, and location of horns are highly diverged within Scarabaeidae, beetle horns are an attractive model for studying the evolution of sexually dimorphic and novel traits. In beetles including the Japanese rhinoceros beetle Trypoxylus dichotomus, the sex differentiation gene doublesex (dsx) plays a crucial role in sexually dimorphic horn formation during larval-pupal development. However, knowledge of when and how dsx drives the gene regulatory network (GRN) for horn formation to form sexually dimorphic horns during development remains elusive. To address this issue, we identified a Trypoxylus-ortholog of the sex determination gene, transformer (tra), that regulates sex-specific splicing of the dsx pre-mRNA, and whose loss of function results in sex transformation. By knocking down tra function at multiple developmental timepoints during larval-pupal development, we estimated the onset when the sex-specific GRN for horn formation is driven. In addition, we also revealed that dsx regulates different aspects of morphogenetic activities during the prepupal and pupal developmental stages to form appropriate morphologies of pupal head and thoracic horn primordia as well as those of adult horns. Based on these findings, we discuss the evolutionary developmental background of sexually dimorphic trait growth in horned beetles.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Coleoptera / genetics*
  • Coleoptera / growth & development*
  • Female
  • Gene Expression Regulation, Developmental
  • Gene Knockdown Techniques
  • Genes, Insect
  • Horns / growth & development
  • Insect Proteins / genetics
  • Larva / genetics
  • Larva / growth & development
  • Male
  • Phenotype
  • Pupa / genetics
  • Pupa / growth & development
  • RNA Interference
  • Sex Characteristics
  • Sex Differentiation / genetics

Substances

  • Insect Proteins

Grants and funding

TN was supported by MEXT KAKENHI (The Ministry of Education, Culture, Sports, Science and Technology; http://www.mext.go.jp/en/) (23128505, 25128706, 16H01452 and 18H04766) and NIBB Cooperative Research Programs (http://www.nibb.ac.jp/en/collabo/collabo.html) (18-433) and NIG-JOINT(https://www.nig.ac.jp/nig/research-infrastructure-collaboration/nig-collaboration-grant-2) (35A2017). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.