Immediate-Early Promoter-Driven Transgenic Reporter System for Neuroethological Research in a Hemimetabolous Insect

Genes expressed in response to increased neuronal activity are widely used as activity markers in recent behavioral neuroscience. In the present study, we established transgenic reporter system for whole-brain activity mapping in the two-spotted cricket , a hemimetabolous insect used in neuroetholog...

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Published ineNeuro Vol. 5; no. 4; p. ENEURO.0061-18.2018
Main Authors Watanabe, Takayuki, Ugajin, Atsushi, Aonuma, Hitoshi
Format Journal Article
LanguageEnglish
Published United States Society for Neuroscience 01.07.2018
Subjects
Methods/New Tools
immediate-early gene
transgenesis
Activity mapping
Gryllus bimaculatus
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Summary:Genes expressed in response to increased neuronal activity are widely used as activity markers in recent behavioral neuroscience. In the present study, we established transgenic reporter system for whole-brain activity mapping in the two-spotted cricket , a hemimetabolous insect used in neuroethology and behavioral ecology. In the cricket brain, a homolog of ( ) was rapidly induced as an immediate-early gene (IEG) in response to neuronal hyperexcitability. The upstream genomic fragment of contains potential binding sites for transcription factors regulated by various intracellular signaling pathways, as well as core promoter elements conserved across insect/crustacean homologs. Using the upstream genomic fragment of we established an IEG promoter-driven transgenic reporter system in the cricket. In the brain of transgenic crickets, the reporter gene (a nuclear-targeted destabilized EYFP) was induced in response to neuronal hyperexcitability. Inducible expression of reporter protein was detected in almost all neurons after neuronal hyperexcitability. Using our novel reporter system, we successfully detected neuronal activation evoked by feeding in the cricket brain. Our IEG promoter-driven activity reporting system allows us to visualize behaviorally relevant neural circuits at cellular resolution in the cricket brain.
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This work was supported, in part, by the Grant-in-Aid for Japanese Society for the Promotion of Science (JSPS) Research Fellow (24•3065) to TW; the JSPS KAKENHI (16K20875) to TW; and Akiyama Life Science Foundation research grant 2014 to TW. The white-eye mutant of G. bimaculatus (gwhite mutant) was a kind gift of Prof. Sumihare Noji in the University of Tokushima. The pXL-BacII, pBSII-IFP2-orf, and pBSII-ITR1.1k-EYFP plasmids were kind gifts of Prof. Malcolm J. Fraser, Jr. in the University of Notre Dame.
Author contributions: T.W. designed the study and wrote the manuscript with the help of A.U. and T.W. performed all experiments and analyzed the data. All the authors reviewed the manuscript.
The authors declare no competing financial interests.
ISSN:2373-2822
2373-2822
DOI:10.1523/eneuro.0061-18.2018