Reference - Detail
| RRC ID | 52750 |
|---|---|
| Author | Hara C, Morishita K, Takayanagi-Kiya S, Mikami A, Uchino K, Sakurai T, Kanzaki R, Sezutsu H, Iwami M, Kiya T. |
| Title | Refinement of ectopic protein expression through the GAL4/UAS system in Bombyx mori: application to behavioral and developmental studies. |
| Journal | Sci Rep |
| 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. |
| Volume | 7(1) |
| Pages | 11795 |
| Published | 2017-9-18 |
| DOI | 10.1038/s41598-017-12102-2 |
| PII | 10.1038/s41598-017-12102-2 |
| PMID | 28924263 |
| PMC | PMC5603595 |
| MeSH | Animals Animals, Genetically Modified* / genetics Animals, Genetically Modified* / metabolism Behavior, Animal* Bombyx* / genetics Bombyx* / metabolism Gene Expression* Insect Proteins* / genetics Insect Proteins* / metabolism Larva / genetics Larva / metabolism Male Recombinant Proteins / biosynthesis Recombinant Proteins / genetics Tetanus Toxin* / biosynthesis Tetanus Toxin* / genetics |
| IF | 3.998 |
| Times Cited | 3 |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | X(Twitter) |
| Total number of mentions | 3 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| DNA material | 20xUAS-IVS-myrGFP-p10T (RDB15812) pBac[20xUAS-IVS-myrGFP-p10T LL-EGFP] (RDB15813) pBac[20xUAS-IVS-GCaMP5G-p10T LL-EGFP] (RDB15814) pBac[20xUAS-IVS-TeTxLC::CFP-p10T LL-EGFP] (RDB15815). |