Endoplasmic Reticulum Calcium Mediates Drosophila Wing Development

Laura Faith George; Mikaela Lynn Follmer; Emily Fontenoy; Hannah Rose Moran; Jeremy Ryan Brown; Yunus H Ozekin; Emily Anne Bates

doi:10.1089/bioe.2022.0036

Endoplasmic Reticulum Calcium Mediates Drosophila Wing Development

Bioelectricity. 2023 Dec 1;5(4):290-306. doi: 10.1089/bioe.2022.0036. Epub 2023 Dec 15.

Authors

Laura Faith George¹, Mikaela Lynn Follmer¹, Emily Fontenoy¹, Hannah Rose Moran¹, Jeremy Ryan Brown¹, Yunus H Ozekin¹, Emily Anne Bates¹

Affiliation

¹ Department of Pediatrics, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA.

PMID: 38143873
PMCID: PMC10733776 (available on 2024-12-01)
DOI: 10.1089/bioe.2022.0036

Abstract

Background: The temporal dynamics of morphogen presentation impacts transcriptional responses and tissue patterning. However, the mechanisms controlling morphogen release are far from clear. We found that inwardly rectifying potassium (Irk) channels regulate endogenous transient increases in intracellular calcium and bone morphogenetic protein (BMP/Dpp) release for Drosophila wing development. Inhibition of Irk channels reduces BMP/Dpp signaling, and ultimately disrupts wing morphology. Ion channels impact development of several tissues and organisms in which BMP signaling is essential. In neurons and pancreatic beta cells, Irk channels modulate membrane potential to affect intracellular Ca⁺⁺ to control secretion of neurotransmitters and insulin. Based on Irk activity in neurons, we hypothesized that electrical activity controls endoplasmic reticulum (ER) Ca⁺⁺ release into the cytoplasm to regulate the release of BMP.

Materials and methods: To test this hypothesis, we reduced expression of four proteins that control ER calcium, Stromal interaction molecule 1 (Stim), Calcium release-activated calcium channel protein 1 (Orai), SarcoEndoplasmic Reticulum Calcium ATPase (SERCA), small conductance calcium-activated potassium channel (SK), and Bestrophin 2 (Best2) using RNAi and documented wing phenotypes. We use live imaging to study calcium and Dpp release within pupal wings and larval wing discs. Additionally, we employed immunohistochemistry to characterize Small Mothers Against Decapentaplegic (SMAD) phosphorylation downstream of the BMP/Dpp pathway following RNAi knockdown.

Results: We found that reduced Stim and SERCA function decreases amplitude and frequency of endogenous calcium transients in the wing disc and reduced BMP/Dpp release.

Conclusion: Our results suggest control of ER calcium homeostasis is required for BMP/Dpp release, and Drosophila wing development.

Keywords: BMP; Drosophila wing development; Orai; SERCA; Stim; bestrophin; bone morphogenic protein; calcium; endoplasmic reticulum; transients; wing venation.