Research Article

Multispectral Imaging and Single-Cell Analysis with Genetically Encoded Biosensors Unveil Complex Interactions between Extracellular ATP and Intracellular Calcium

Volume: 45 Number: 2 June 30, 2024
EN

Multispectral Imaging and Single-Cell Analysis with Genetically Encoded Biosensors Unveil Complex Interactions between Extracellular ATP and Intracellular Calcium

Abstract

The dynamic interplay between extracellular ATP (eATP) and intracellular calcium ([Ca2+]i) serves as a pivotal signaling axis in cellular physiology, influencing a myriad of cellular processes. Traditionally recognized as an energy currency within the cell, ATP has emerged as a multifunctional signaling molecule that orchestrates diverse cellular responses through activation of purinergic receptors. The complex link between ATP signaling and calcium dynamics plays a central role in cellular communication and homeostasis. Advancements in imaging technologies such as development of genetically encoded biosensors have revolutionized the study of cellular signaling dynamics, enabling visualization of the spatiotemporal aspects of eATP and [Ca2+]i in real-time. The convergence of eATP signaling and [Ca2+]i dynamics serves as a central hub in cellular communication. In this study, utilizing bicistronic construct biosensors for multispectral imaging of [Ca2+]i responses to eATP, we show that distinct concentrations of eATP administration reveal complex intracellular [Ca2+]i responses, potentially attributed to receptor desensitization. Single-cell co-imaging uncovers [Ca2+]i heterogeneity, emphasizing the significance of individual cell dynamics in eATP-induced calcium signaling. Therefore, this study sheds light on the intricacies of eATP-induced calcium signaling, providing insights valuable for basic research and therapeutic applications.

Keywords

Project Number

EMBO IG-5322-2023

Ethical Statement

N.A.

Thanks

This research was supported by funds from the European Molecular Biology Organization EMBO (IG-5322-2023. The author acknowledges Asal Ghaffari Zaki for English language proofreading. Figure 1A was generated with Biorender (Licence code: ME26EKR3WY).

References

  1. [1] Arcuino G, Lin JHC, Takano T, Liu C, Jiang L, Gao Q, Kang J, Nedergaard M. Intercellular calcium signaling mediated by point-source burst release of ATP, Proc. Natl. Acad. Sci., 99 (15) (2002) 9840–9845.
  2. [2] Cisneros-Mejorado A, Pérez-Samartín A, Gottlieb M, Matute C. ATP signaling in brain: release, excitotoxicity and potential therapeutic targets, Cell. Mol. Neurobiol., 35 (1) (2015) 1–6.
  3. [3] Depaoli MR, Karsten F, Madreiter-Sokolowski CT, Klec C, Gottschalk B, Bischof H, Eroglu E, Waldeck-Weiermair M, Simmen T, Graier WF, Malli R. Real-Time Imaging of Mitochondrial ATP Dynamics Reveals the Metabolic Setting of Single Cells, Cell Rep., 25 (2) (2018) 501-512.
  4. [4] Bonora M, Patergnani S, Rimessi A, De Marchi E, Suski JM, Bononi A, Giorgi C, Marchi S, Missiroli S, Poletti F, Wieckowski MR, Pinton P., ATP synthesis and storage, Purinergic Signal., 8 (3) (2012) 343–357.
  5. [5] Eersapah V, Hugel S, Schlichter R., High-resolution detection of ATP release from single cultured mouse dorsal horn spinal cord glial cells and its modulation by noradrenaline, Purinergic Signal., 15 (3) (2019) 403–420.
  6. [6] Fields RD, Burnstock G., Purinergic signalling in neuron-glia interactions, Nat. Rev. Neurosci., 7 (6) (2006) 423–436.
  7. [7] Eroglu E, Saravi SSS, Sorrentino A, Steinhorn B, Michel T., Discordance between eNOS phosphorylation and activation revealed by multispectral imaging and chemogenetic methods, Proc. Natl. Acad. Sci. U. S. A., 116 (40) (2019) 20210–20217.
  8. [8] Eroglu E, Hallström S, Bischof H, Opelt M, Schmidt K, Mayer B, Waldeck-Weiermair M, Graier WF, Malli R., Real-time visualization of distinct nitric oxide generation of nitric oxide synthase isoforms in single cells, Nitric Oxide Biol. Chem., 70 (2017) 59–67.

Details

Primary Language

English

Subjects

Signal Transduction

Journal Section

Research Article

Publication Date

June 30, 2024

Submission Date

April 1, 2024

Acceptance Date

June 7, 2024

Published in Issue

Year 2024 Volume: 45 Number: 2

APA
Eroğlu, E. (2024). Multispectral Imaging and Single-Cell Analysis with Genetically Encoded Biosensors Unveil Complex Interactions between Extracellular ATP and Intracellular Calcium. Cumhuriyet Science Journal, 45(2), 360-365. https://doi.org/10.17776/csj.1462654

As of 2026, Cumhuriyet Science Journal will be published in six issues per year, released in February, April, June, August, October, and December