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Sci Rep

Title:		Refinement of ectopic protein expression through the GAL4/UAS system in Bombyx mori: application to behavioral and developmental studies
Author(s):		Hara C; Morishita K; Takayanagi-Kiya S; Mikami A; Uchino K; Sakurai T; Kanzaki R; Sezutsu H; Iwami M; Kiya T;
Address:		"Division of Life Sciences, Graduate School of Natural Science and Technology, Kanazawa University, Kakuma-machi, Kanazawa, Ishikawa, 920-1192, Japan. Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo, 153-8904, Japan. Transgenic Silkworm Research Unit, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, 1-2 Owashi, Tsukuba, Ibaraki, 305-8634, Japan. Division of Life Sciences, Graduate School of Natural Science and Technology, Kanazawa University, Kakuma-machi, Kanazawa, Ishikawa, 920-1192, Japan. kiya@staff.kanazawa-u.ac.jp"
Journal Title:		Sci Rep
Year:		2017
Volume:		20170918
Issue:		1
Page Number:		11795 -
DOI:		10.1038/s41598-017-12102-2
ISSN/ISBN:		2045-2322 (Electronic) 2045-2322 (Linking)
Abstract:		"Silkmoth, Bombyx mori, is one of the important model insects in which transgenic techniques and the GAL4/UAS system are applicable. However, due to cytotoxicity and low transactivation activity of GAL4, effectiveness of the GAL4/UAS system and its application in B. mori are still limited. In the present study, we refined the previously reported UAS vector by exploiting transcriptional and translational enhancers, and achieved 200-fold enhancement of reporter GFP fluorescence in the GAL4/UAS system. Enhanced protein expression of membrane-targeted GFP and calcium indicator protein (GCaMP5G) drastically improved visualization of fine neurite structures and neural activity, respectively. Also, with the refined system, we generated a transgenic strain that expresses tetanus toxin light chain (TeTxLC), which blocks synaptic transmission, under the control of GAL4. Ectopic TeTxLC expression in the sex pheromone receptor neurons inhibited male courtship behavior, proving effectiveness of TeTxLC on loss-of-function analyses of neural circuits. In addition, suppression of prothoracicotropic hormone (PTTH) or insulin-like peptide (bombyxin) secretion impaired developmental timing and growth rate, respectively. Furthermore, we revealed that larval growth is sex-differentially regulated by these peptide hormones. The present study provides important technical underpinnings of transgenic approaches in silkmoths and insights into mechanisms of postembryonic development in insects"
Keywords:		"Animals *Animals, Genetically Modified/genetics/metabolism *Behavior, Animal *Bombyx/genetics/metabolism *Gene Expression *Insect Proteins/genetics/metabolism Larva/genetics/metabolism Male Recombinant Proteins/biosynthesis/genetics *Tetanus Toxin/biosynt;"
Notes:		"MedlineHara, Chiho Morishita, Koudai Takayanagi-Kiya, Seika Mikami, Akihisa Uchino, Keiro Sakurai, Takeshi Kanzaki, Ryohei Sezutsu, Hideki Iwami, Masafumi Kiya, Taketoshi eng Wellcome Trust/United Kingdom Research Support, Non-U.S. Gov't England 2017/09/20 Sci Rep. 2017 Sep 18; 7(1):11795. doi: 10.1038/s41598-017-12102-2"