Get Closer to the World of Contact Sites: A Beginner's Guide to Proximity-Driven Fluorescent Probes. (December 2022)

• Record Type: Journal Article
• Authors: Poggio, Elena; Brini, Marisa; Calì, Tito
• Is Part Of: Contact. Volume 5(2022)
• Publication Date: 2022-12
• Subjects: organelle contact sites -- SPLICS -- FRET -- PLA -- BiFC; Cell interaction -- Periodicals; Cells -- Periodicals; Cytology -- Periodicals; 571.6
• DOI: 10.1177/25152564221135748
• Languages: English
• ISSNs: 2515-2564

Abstract

To maintain cellular homeostasis and to coordinate the proper response to a specific stimulus, information must be integrated throughout the cell in a well-organized network, in which organelles are the crucial nodes and membrane contact sites are the main edges. Membrane contact sites are the cellular subdomains where two or more organelles come into close apposition and interact with each other. Even though many inter-organelle contacts have been identified, most of them are still not fully characterized, therefore their study is an appealing and expanding field of research. Thanks to significant technological progress, many tools are now available or are in rapid development, making it difficult to choose which one is the most suitable for answering a specific biological question. Here we distinguish two different experimental approaches for studying inter-organelle contact sites. The first one aims to morphologically characterize the sites of membrane contact and to identify the molecular players involved, relying mainly on the application of biochemical and electron microscopy (EM)-related methods. The second approach aims to understand the functional importance of a specific contact, focusing on spatio-temporal details. For this purpose, proximity-driven fluorescent probes are the experimental tools of choice, since they allow the monitoring and quantification of membrane contact sites and their dynamics in living cells under different cellular conditions or upon different stimuli. In this review, we focus on these tools with the purpose of highlighting their great versatility and how they can be applied in the study of membrane contacts. We will extensively describe all the different types of proximity-driven fluorescent tools, discussing their benefits and drawbacks, ultimately providing some suggestions to choose and apply the appropriate methods on a case-to-case basis and to obtain the best experimental outcomes.