Quantitative analysis of lipid droplet fusion: inefficient steady state fusion but rapid stimulation by chemical fusogens
Lipid droplets (LDs) are dynamic cytoplasmic organelles containing neutral lipids and bounded by a phospholipid monolayer. Previous studies have suggested that LDs can undergo constitutive homotypic fusion, a process linked to the inhibitory effects of fatty acids on glucose transporter trafficking....
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| Veröffentlicht in: | PloS one 2010-12, Vol.5 (12), p.e15030-e15030 |
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| description | Lipid droplets (LDs) are dynamic cytoplasmic organelles containing neutral lipids and bounded by a phospholipid monolayer. Previous studies have suggested that LDs can undergo constitutive homotypic fusion, a process linked to the inhibitory effects of fatty acids on glucose transporter trafficking. Using strict quantitative criteria for LD fusion together with refined light microscopic methods and real-time analysis, we now show that LDs in diverse cell types show low constitutive fusogenic activity under normal growth conditions. To investigate the possible modulation of LD fusion, we screened for agents that can trigger fusion. A number of pharmacological agents caused homotypic fusion of lipid droplets in a variety of cell types. This provided a novel cell system to study rapid regulated fusion between homotypic phospholipid monolayers. LD fusion involved an initial step in which the two adjacent membranes became continuous ( |
| doi_str_mv | 10.1371/journal.pone.0015030 |
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Previous studies have suggested that LDs can undergo constitutive homotypic fusion, a process linked to the inhibitory effects of fatty acids on glucose transporter trafficking. Using strict quantitative criteria for LD fusion together with refined light microscopic methods and real-time analysis, we now show that LDs in diverse cell types show low constitutive fusogenic activity under normal growth conditions. To investigate the possible modulation of LD fusion, we screened for agents that can trigger fusion. A number of pharmacological agents caused homotypic fusion of lipid droplets in a variety of cell types. This provided a novel cell system to study rapid regulated fusion between homotypic phospholipid monolayers. LD fusion involved an initial step in which the two adjacent membranes became continuous (<10 s), followed by the slower merging (100 s) of the neutral lipid cores to produce a single spherical LD. These fusion events were accompanied by changes to the LD surface organization. Measurements of LDs undergoing homotypic fusion showed that fused LDs maintained their initial volume, with a corresponding decrease in surface area suggesting rapid removal of membrane from the fused LD. This study provides estimates for the level of constitutive LD fusion in cells and questions the role of LD fusion in vivo. In addition, it highlights the extent of LD restructuring which occurs when homotypic LD fusion is triggered in a variety of cell types.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0015030</identifier><identifier>PMID: 21203462</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>3T3 Cells ; Adipocytes ; Adipocytes - cytology ; Animals ; Biology ; Cores ; Cricetinae ; Cytoplasm - metabolism ; Droplets ; Fatty acids ; Fibroblasts ; Fibroblasts - cytology ; Fibroblasts - metabolism ; Fluorescent Antibody Technique, Indirect - methods ; Free radicals ; Glucose transporter ; Glycerol ; Growth conditions ; Humans ; Isoquinolines - pharmacology ; Kinases ; Lipid Metabolism ; Lipid peroxidation ; Lipids ; Lipids - chemistry ; Membranes ; Mice ; Microscopy ; Microscopy - methods ; Models, Theoretical ; Monolayers ; Monomolecular films ; Morphology ; Organelles ; Pharmacology ; Phospholipids ; Phospholipids - chemistry ; Protein Kinase Inhibitors - pharmacology ; Proteins ; Quantitative analysis ; Studies ; Sulfonamides - pharmacology</subject><ispartof>PloS one, 2010-12, Vol.5 (12), p.e15030-e15030</ispartof><rights>COPYRIGHT 2010 Public Library of Science</rights><rights>2010 Murphy et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 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Previous studies have suggested that LDs can undergo constitutive homotypic fusion, a process linked to the inhibitory effects of fatty acids on glucose transporter trafficking. Using strict quantitative criteria for LD fusion together with refined light microscopic methods and real-time analysis, we now show that LDs in diverse cell types show low constitutive fusogenic activity under normal growth conditions. To investigate the possible modulation of LD fusion, we screened for agents that can trigger fusion. A number of pharmacological agents caused homotypic fusion of lipid droplets in a variety of cell types. This provided a novel cell system to study rapid regulated fusion between homotypic phospholipid monolayers. LD fusion involved an initial step in which the two adjacent membranes became continuous (<10 s), followed by the slower merging (100 s) of the neutral lipid cores to produce a single spherical LD. 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In addition, it highlights the extent of LD restructuring which occurs when homotypic LD fusion is triggered in a variety of cell types.</description><subject>3T3 Cells</subject><subject>Adipocytes</subject><subject>Adipocytes - cytology</subject><subject>Animals</subject><subject>Biology</subject><subject>Cores</subject><subject>Cricetinae</subject><subject>Cytoplasm - metabolism</subject><subject>Droplets</subject><subject>Fatty acids</subject><subject>Fibroblasts</subject><subject>Fibroblasts - cytology</subject><subject>Fibroblasts - metabolism</subject><subject>Fluorescent Antibody Technique, Indirect - methods</subject><subject>Free radicals</subject><subject>Glucose transporter</subject><subject>Glycerol</subject><subject>Growth conditions</subject><subject>Humans</subject><subject>Isoquinolines - pharmacology</subject><subject>Kinases</subject><subject>Lipid Metabolism</subject><subject>Lipid peroxidation</subject><subject>Lipids</subject><subject>Lipids - chemistry</subject><subject>Membranes</subject><subject>Mice</subject><subject>Microscopy</subject><subject>Microscopy - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Murphy, Samantha</au><au>Martin, Sally</au><au>Parton, Robert G</au><au>Gruenberg, Jean</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Quantitative analysis of lipid droplet fusion: inefficient steady state fusion but rapid stimulation by chemical fusogens</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2010-12-23</date><risdate>2010</risdate><volume>5</volume><issue>12</issue><spage>e15030</spage><epage>e15030</epage><pages>e15030-e15030</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Lipid droplets (LDs) are dynamic cytoplasmic organelles containing neutral lipids and bounded by a phospholipid monolayer. Previous studies have suggested that LDs can undergo constitutive homotypic fusion, a process linked to the inhibitory effects of fatty acids on glucose transporter trafficking. Using strict quantitative criteria for LD fusion together with refined light microscopic methods and real-time analysis, we now show that LDs in diverse cell types show low constitutive fusogenic activity under normal growth conditions. To investigate the possible modulation of LD fusion, we screened for agents that can trigger fusion. A number of pharmacological agents caused homotypic fusion of lipid droplets in a variety of cell types. This provided a novel cell system to study rapid regulated fusion between homotypic phospholipid monolayers. LD fusion involved an initial step in which the two adjacent membranes became continuous (<10 s), followed by the slower merging (100 s) of the neutral lipid cores to produce a single spherical LD. These fusion events were accompanied by changes to the LD surface organization. Measurements of LDs undergoing homotypic fusion showed that fused LDs maintained their initial volume, with a corresponding decrease in surface area suggesting rapid removal of membrane from the fused LD. This study provides estimates for the level of constitutive LD fusion in cells and questions the role of LD fusion in vivo. In addition, it highlights the extent of LD restructuring which occurs when homotypic LD fusion is triggered in a variety of cell types.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>21203462</pmid><doi>10.1371/journal.pone.0015030</doi><tpages>e15030</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | 3T3 Cells Adipocytes Adipocytes - cytology Animals Biology Cores Cricetinae Cytoplasm - metabolism Droplets Fatty acids Fibroblasts Fibroblasts - cytology Fibroblasts - metabolism Fluorescent Antibody Technique, Indirect - methods Free radicals Glucose transporter Glycerol Growth conditions Humans Isoquinolines - pharmacology Kinases Lipid Metabolism Lipid peroxidation Lipids Lipids - chemistry Membranes Mice Microscopy Microscopy - methods Models, Theoretical Monolayers Monomolecular films Morphology Organelles Pharmacology Phospholipids Phospholipids - chemistry Protein Kinase Inhibitors - pharmacology Proteins Quantitative analysis Studies Sulfonamides - pharmacology |
| title | Quantitative analysis of lipid droplet fusion: inefficient steady state fusion but rapid stimulation by chemical fusogens |
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