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利用红外激光热休克操纵基因表达及其在果蝇气管发育研究中的应用。

Manipulation of gene expression by infrared laser heat shock and its application to the study of tracheal development in Drosophila.

作者信息

Miao Guangxia, Hayashi Shigeo

机构信息

Laboratory for Morphogenetic Signaling, RIKEN Center for Developmental Biology, Chuo-ku, Kobe, Hyogo, Japan; Department of Biology, Kobe University Graduate School of Science, Nada-ku, Kobe, Hyogo, Japan.

出版信息

Dev Dyn. 2015 Mar;244(3):479-87. doi: 10.1002/dvdy.24192.

Abstract

BACKGROUND

Induction of gene expression in a specific cell and a defined time window is desirable to investigate gene function at the cellular level during morphogenesis. To achieve this, we attempted to introduce the infrared laser-evoked gene operator system (IR-LEGO, Kamei et al., 2009) in the Drosophila embryo. In this technique, infrared laser light illumination induces genes to be expressed under the control of heat shock promoters at the single cell level.

RESULTS

We applied IR-LEGO to a transgenic fly stock, HS-eGFP, in which the enhanced green fluorescent protein (eGFP) gene is placed under the control of heat shock protein 70 promoter, and showed that eGFP expression can be induced in single cells within 1-2 hr after IR illumination. Furthermore, induction of HS-Branchless transgene encoding the Drosophila fibroblast growth factor (FGF) effectively altered the migration and branching patterns of the tracheal system.

CONCLUSIONS

Our results indicated that IR-LEGO is a promising choice for the timely control of gene expression in a small group of cells in the Drosophila embryo. By using IR-LEGO, we further demonstrated that the tracheal terminal branching program is sensitive to localized expression of exogenous FGF.

摘要

背景

在特定细胞和限定的时间窗口内诱导基因表达,对于在形态发生过程中在细胞水平研究基因功能是很有必要的。为实现这一点,我们尝试在果蝇胚胎中引入红外激光诱发基因操纵系统(IR-LEGO,龟井等人,2009年)。在这项技术中,红外激光照射可诱导基因在单细胞水平上在热休克启动子的控制下表达。

结果

我们将IR-LEGO应用于一种转基因果蝇品系HS-eGFP,其中增强型绿色荧光蛋白(eGFP)基因置于热休克蛋白70启动子的控制之下,结果表明在红外照射后1-2小时内可在单细胞中诱导eGFP表达。此外,编码果蝇成纤维细胞生长因子(FGF)的HS-Branchless转基因的诱导有效地改变了气管系统的迁移和分支模式。

结论

我们的结果表明,IR-LEGO是在果蝇胚胎中一小群细胞中及时控制基因表达的一个有前景的选择。通过使用IR-LEGO,我们进一步证明气管末端分支程序对外源FGF的局部表达敏感。

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