Methods for making and observing model lipid droplets.

Autor:	Gandhi SA; Department of Physics and Astronomy, Wayne State University, Detroit, MI, USA 48201., Parveen S; Department of Physics and Astronomy, Wayne State University, Detroit, MI, USA 48201., Alduhailan M; Department of Physics and Astronomy, Wayne State University, Detroit, MI, USA 48201., Tripathi R; Department of Physics and Astronomy, Wayne State University, Detroit, MI, USA 48201., Junedi N; Department of Physics and Astronomy, Wayne State University, Detroit, MI, USA 48201., Saqallah M; Department of Physics and Astronomy, Wayne State University, Detroit, MI, USA 48201., Sanders MA; Center for Molecular Medicine and Genetics, School of Medicine, Wayne State University, Detroit, MI, USA 40201.; Center for Integrative Metabolic and Endocrine Research, School of Medicine, Wayne State University, Detroit, MI USA 48201., Hoffmann PM; Department of Physics and Astronomy, Wayne State University, Detroit, MI, USA 48201.; Physical Sciences Department, Embry-Riddle Aeronautical University, Daytona Beach, FL, USA 32114., Truex K; Department of Physics, United States Naval Academy, Annapolis, MD, USA 21402., Granneman JG; Center for Molecular Medicine and Genetics, School of Medicine, Wayne State University, Detroit, MI, USA 40201.; Center for Integrative Metabolic and Endocrine Research, School of Medicine, Wayne State University, Detroit, MI USA 48201., Kelly CV; Department of Physics and Astronomy, Wayne State University, Detroit, MI, USA 48201.; Center for Integrative Metabolic and Endocrine Research, School of Medicine, Wayne State University, Detroit, MI USA 48201.
Jazyk:	angličtina
Zdroj:	BioRxiv : the preprint server for biology [bioRxiv] 2023 Jul 19. Date of Electronic Publication: 2023 Jul 19.
DOI:	10.1101/2023.07.17.549385
Abstrakt:	The mechanisms by which the lipid droplet (LD) membrane is remodeled in concert with the activation of lipolysis incorporate a complex interplay of proteins, phospholipids, and neutral lipids. Model LDs (mLDs) provide an isolated, purified system for testing the mechanisms by which the droplet composition, size, shape, and tension affects triglyceride metabolism. Described here are methods of making and testing mLDs ranging from 0.1 to 40 μm diameter with known composition. Methods are described for imaging mLDs with high-resolution microscopy during buffer exchanges for the measurement of membrane binding, diffusion, and tension via fluorescence correlation spectroscopy (FCS), fluorescence recovery after photobleaching (FRAP), fluorescence lifetime imaging microscopy (FLIM), atomic force microscopy (AFM), pendant droplet tensiometry, and imaging flow cytometry. These complementary, cross-validating methods of measuring LD membrane behavior reveal the interplay of biophysical processes in triglyceride metabolism. Competing Interests: Declaration of Interests The authors declare no conflicts of interest.
Databáze:	MEDLINE
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