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An Optogenetic Method to Study Signal Transduction in Intestinal Stem Cell Homeostasis.
Journal of Molecular Biology ( IF 4.7 ) Pub Date : 2020-03-20 , DOI: 10.1016/j.jmb.2020.03.019 Nawat Bunnag 1 , Qian Hui Tan 1 , Prameet Kaur 1 , Anupriya Ramamoorthy 1 , Isabelle Chiao Han Sung 1 , Jay Lusk 1 , Nicholas S Tolwinski 1
Journal of Molecular Biology ( IF 4.7 ) Pub Date : 2020-03-20 , DOI: 10.1016/j.jmb.2020.03.019 Nawat Bunnag 1 , Qian Hui Tan 1 , Prameet Kaur 1 , Anupriya Ramamoorthy 1 , Isabelle Chiao Han Sung 1 , Jay Lusk 1 , Nicholas S Tolwinski 1
Affiliation
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Homeostasis in adult organs involves replacement of cells from a stem cell pool maintained in specialized niches regulated by extracellular signals. This cell-to-cell communication employs signal transduction pathways allowing cells to respond with a variety of behaviors. To study these cellular behaviors, signaling must be perturbed within tissues in precise patterns, a technique recently made possible by the development of optogenetic tools. We developed tools to study signal transduction in vivo in an adult fly midgut stem cell model where signaling was regulated by the application of light. Activation was achieved by clustering of membrane receptors EGFR and Toll, while inactivation was achieved by clustering the downstream activators ERK/Rolled and NFκB/Dorsal in the cytoplasm, preventing nuclear translocation and transcriptional activation. We show that both pathways contribute to stem and transit amplifying cell numbers and affect the lifespan of adult flies. We further present new approaches to overcome overexpression phenotypes and novel methods for the integration of optogenetics into the already-established genetic toolkit of Drosophila.
中文翻译：
研究肠道干细胞稳态的信号转导的光遗传学方法。
成年器官的体内平衡涉及从干细胞库中置换细胞，该干细胞库维持在受细胞外信号调节的特殊壁ches中。这种细胞间通信采用信号转导途径，使细胞能够以多种行为做出反应。为了研究这些细胞行为，必须以精确的模式在组织内扰动信号传导，最近通过光遗传学工具的发展使该技术成为可能。我们开发了用于研究成年蝇中肠干细胞模型体内信号转导的工具，该模型通过光的作用来调节信号传导。激活是通过聚集膜受体EGFR和Toll来实现的，而灭活是通过将下游激活剂ERK / Rolled和NFκB/ Dorsal聚集在细胞质中来实现的，从而防止了核移位和转录激活。我们表明这两种途径有助于干和转运放大细胞数量，并影响成年苍蝇的寿命。我们进一步提出了克服过度表达表型的新方法，以及将光遗传学整合到果蝇已经建立的遗传工具包中的新方法。
更新日期：2020-03-20
中文翻译：
研究肠道干细胞稳态的信号转导的光遗传学方法。
成年器官的体内平衡涉及从干细胞库中置换细胞，该干细胞库维持在受细胞外信号调节的特殊壁ches中。这种细胞间通信采用信号转导途径，使细胞能够以多种行为做出反应。为了研究这些细胞行为，必须以精确的模式在组织内扰动信号传导，最近通过光遗传学工具的发展使该技术成为可能。我们开发了用于研究成年蝇中肠干细胞模型体内信号转导的工具，该模型通过光的作用来调节信号传导。激活是通过聚集膜受体EGFR和Toll来实现的，而灭活是通过将下游激活剂ERK / Rolled和NFκB/ Dorsal聚集在细胞质中来实现的，从而防止了核移位和转录激活。我们表明这两种途径有助于干和转运放大细胞数量，并影响成年苍蝇的寿命。我们进一步提出了克服过度表达表型的新方法，以及将光遗传学整合到果蝇已经建立的遗传工具包中的新方法。
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