Rapid Assessment of Insect Steroid Hormone Entry Into Cultured Cells

Mitchell Masterson; Riyan Bittar; Riyan Bittar; Hannah Chu; Naoki Yamanaka; Sachiko Haga-Yamanaka

doi:10.3389/fphys.2021.816058

Frontiers in Physiology (Jan 2022)

Rapid Assessment of Insect Steroid Hormone Entry Into Cultured Cells

Mitchell Masterson,
Riyan Bittar,
Riyan Bittar,
Hannah Chu,
Naoki Yamanaka,
Sachiko Haga-Yamanaka

Affiliations

Mitchell Masterson: Department of Molecular, Cell and Systems Biology, University of California, Riverside, Riverside, CA, United States
Riyan Bittar: Department of Molecular, Cell and Systems Biology, University of California, Riverside, Riverside, CA, United States
Riyan Bittar: Department of Entomology, University of California, Riverside, Riverside, CA, United States
Hannah Chu: Department of Entomology, University of California, Riverside, Riverside, CA, United States
Naoki Yamanaka: Department of Entomology, University of California, Riverside, Riverside, CA, United States
Sachiko Haga-Yamanaka: Department of Molecular, Cell and Systems Biology, University of California, Riverside, Riverside, CA, United States

DOI: https://doi.org/10.3389/fphys.2021.816058
Journal volume & issue: Vol. 12

Abstract

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Steroid hormones control development and homeostasis in a wide variety of animals by interacting with intracellular nuclear receptors. Recent discoveries in the fruit fly Drosophila melanogaster revealed that insect steroid hormones or ecdysteroids are incorporated into cells through a membrane transporter named Ecdysone Importer (EcI), which may become a novel target for manipulating steroid hormone signaling in insects. In this study, we established an assay system that can rapidly assess EcI-mediated ecdysteroid entry into cultured cells. Using NanoLuc Binary Technology (NanoBiT), we first developed an assay to detect ligand-dependent heterodimerization of the ecdysone receptor (EcR) and retinoid X receptor (RXR) in human embryonic kidney (HEK) 293T cells. We also developed HEK293 cells that stably express EcI. By combining these tools, we can monitor ecdysteroid entry into the cells in real time, making it a reliable system to assess EcI-mediated steroid hormone incorporation into animal cells.

Published in Frontiers in Physiology

ISSN: 1664-042X (Online)
Publisher: Frontiers Media S.A.
Country of publisher: Switzerland
LCC subjects: Science: Physiology
Website: https://www.frontiersin.org/journals/physiology

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