Establishment of tools for neurogenetic analysis of sexual behavior in the silkmoth, Bombyx mori.

Silkmoth, Bombyx mori, is an ideal model insect for investigating the neural mechanisms underlying sex pheromone-induced innate behavior. Although transgenic techniques and the GAL4/UAS system are well established in the silkmoth, genetic tools useful for investigating brain function at the neural c...

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Main Authors: Taketoshi Kiya, Koudai Morishita, Keiro Uchino, Masafumi Iwami, Hideki Sezutsu
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2014-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC4232604?pdf=render
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spelling doaj-f5447d5713c54ef7ad0bf214ba22b8222020-11-24T21:50:57ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-01911e11315610.1371/journal.pone.0113156Establishment of tools for neurogenetic analysis of sexual behavior in the silkmoth, Bombyx mori.Taketoshi KiyaKoudai MorishitaKeiro UchinoMasafumi IwamiHideki SezutsuSilkmoth, Bombyx mori, is an ideal model insect for investigating the neural mechanisms underlying sex pheromone-induced innate behavior. Although transgenic techniques and the GAL4/UAS system are well established in the silkmoth, genetic tools useful for investigating brain function at the neural circuit level have been lacking.In the present study, we established silkmoth strains in which we could visualize neural projections (UAS-mCD8GFP) and cell nucleus positions (UAS-GFP.nls), and manipulate neural excitability by thermal stimulation (UAS-dTrpA1). In these strains, neural projections and nucleus position were reliably labeled with green fluorescent protein in a GAL4-dependent manner. Further, the behavior of silkworm larvae and adults could be controlled by GAL4-dependent misexpression of dTrpA1. Ubiquitous dTrpA1 misexpression led both silkmoth larvae and adults to exhibit seizure-like phenotypes in a heat stimulation-dependent manner. Furthermore, dTrpA1 misexpression in the sex pheromone receptor neurons of male silkmoths allowed us to control male sexual behavior by changing the temperature. Thermally stimulated male silkmoths exhibited full sexual behavior, including wing-flapping, orientation, and attempted copulation, and precisely approached a thermal source in a manner similar to male silkmoths stimulated with the sex pheromone.These findings indicate that a thermogenetic approach using dTrpA1 is feasible in Lepidopteran insects and thermogenetic analysis of innate behavior is applicable in the silkmoth. These tools are essential for elucidating the relationships between neural circuits and function using neurogenetic methods.http://europepmc.org/articles/PMC4232604?pdf=render
collection DOAJ
language English
format Article
sources DOAJ
author Taketoshi Kiya
Koudai Morishita
Keiro Uchino
Masafumi Iwami
Hideki Sezutsu
spellingShingle Taketoshi Kiya
Koudai Morishita
Keiro Uchino
Masafumi Iwami
Hideki Sezutsu
Establishment of tools for neurogenetic analysis of sexual behavior in the silkmoth, Bombyx mori.
PLoS ONE
author_facet Taketoshi Kiya
Koudai Morishita
Keiro Uchino
Masafumi Iwami
Hideki Sezutsu
author_sort Taketoshi Kiya
title Establishment of tools for neurogenetic analysis of sexual behavior in the silkmoth, Bombyx mori.
title_short Establishment of tools for neurogenetic analysis of sexual behavior in the silkmoth, Bombyx mori.
title_full Establishment of tools for neurogenetic analysis of sexual behavior in the silkmoth, Bombyx mori.
title_fullStr Establishment of tools for neurogenetic analysis of sexual behavior in the silkmoth, Bombyx mori.
title_full_unstemmed Establishment of tools for neurogenetic analysis of sexual behavior in the silkmoth, Bombyx mori.
title_sort establishment of tools for neurogenetic analysis of sexual behavior in the silkmoth, bombyx mori.
publisher Public Library of Science (PLoS)
series PLoS ONE
issn 1932-6203
publishDate 2014-01-01
description Silkmoth, Bombyx mori, is an ideal model insect for investigating the neural mechanisms underlying sex pheromone-induced innate behavior. Although transgenic techniques and the GAL4/UAS system are well established in the silkmoth, genetic tools useful for investigating brain function at the neural circuit level have been lacking.In the present study, we established silkmoth strains in which we could visualize neural projections (UAS-mCD8GFP) and cell nucleus positions (UAS-GFP.nls), and manipulate neural excitability by thermal stimulation (UAS-dTrpA1). In these strains, neural projections and nucleus position were reliably labeled with green fluorescent protein in a GAL4-dependent manner. Further, the behavior of silkworm larvae and adults could be controlled by GAL4-dependent misexpression of dTrpA1. Ubiquitous dTrpA1 misexpression led both silkmoth larvae and adults to exhibit seizure-like phenotypes in a heat stimulation-dependent manner. Furthermore, dTrpA1 misexpression in the sex pheromone receptor neurons of male silkmoths allowed us to control male sexual behavior by changing the temperature. Thermally stimulated male silkmoths exhibited full sexual behavior, including wing-flapping, orientation, and attempted copulation, and precisely approached a thermal source in a manner similar to male silkmoths stimulated with the sex pheromone.These findings indicate that a thermogenetic approach using dTrpA1 is feasible in Lepidopteran insects and thermogenetic analysis of innate behavior is applicable in the silkmoth. These tools are essential for elucidating the relationships between neural circuits and function using neurogenetic methods.
url http://europepmc.org/articles/PMC4232604?pdf=render
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