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An Optogenetic Method to Study Signal Transduction in Intestinal Stem Cell Homeostasis.

Publication ,  Journal Article
Bunnag, N; Tan, QH; Kaur, P; Ramamoorthy, A; Sung, ICH; Lusk, J; Tolwinski, NS
Published in: Journal of molecular biology
May 2020

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.

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Published In

Journal of molecular biology

DOI

EISSN

1089-8638

ISSN

0022-2836

Publication Date

May 2020

Volume

432

Issue

10

Start / End Page

3159 / 3176

Related Subject Headings

  • Stem Cells
  • Signal Transduction
  • Optogenetics
  • Longevity
  • Light
  • Intestinal Mucosa
  • Homeostasis
  • Gene Regulatory Networks
  • Gene Expression Regulation
  • Drosophila melanogaster
 

Citation

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Bunnag, N., Tan, Q. H., Kaur, P., Ramamoorthy, A., Sung, I. C. H., Lusk, J., & Tolwinski, N. S. (2020). An Optogenetic Method to Study Signal Transduction in Intestinal Stem Cell Homeostasis. Journal of Molecular Biology, 432(10), 3159–3176. https://doi.org/10.1016/j.jmb.2020.03.019
Bunnag, Nawat, Qian Hui Tan, Prameet Kaur, Anupriya Ramamoorthy, Isabelle Chiao Han Sung, Jay Lusk, and Nicholas S. Tolwinski. “An Optogenetic Method to Study Signal Transduction in Intestinal Stem Cell Homeostasis.Journal of Molecular Biology 432, no. 10 (May 2020): 3159–76. https://doi.org/10.1016/j.jmb.2020.03.019.
Bunnag N, Tan QH, Kaur P, Ramamoorthy A, Sung ICH, Lusk J, et al. An Optogenetic Method to Study Signal Transduction in Intestinal Stem Cell Homeostasis. Journal of molecular biology. 2020 May;432(10):3159–76.
Bunnag, Nawat, et al. “An Optogenetic Method to Study Signal Transduction in Intestinal Stem Cell Homeostasis.Journal of Molecular Biology, vol. 432, no. 10, May 2020, pp. 3159–76. Epmc, doi:10.1016/j.jmb.2020.03.019.
Bunnag N, Tan QH, Kaur P, Ramamoorthy A, Sung ICH, Lusk J, Tolwinski NS. An Optogenetic Method to Study Signal Transduction in Intestinal Stem Cell Homeostasis. Journal of molecular biology. 2020 May;432(10):3159–3176.
Journal cover image

Published In

Journal of molecular biology

DOI

EISSN

1089-8638

ISSN

0022-2836

Publication Date

May 2020

Volume

432

Issue

10

Start / End Page

3159 / 3176

Related Subject Headings

  • Stem Cells
  • Signal Transduction
  • Optogenetics
  • Longevity
  • Light
  • Intestinal Mucosa
  • Homeostasis
  • Gene Regulatory Networks
  • Gene Expression Regulation
  • Drosophila melanogaster