Heterogeneity in the physiological states and pharmacological responses of differentiating 3T3-L1 preadipocytes
Increases in key components of adipogenesis and lipolysis pathways correlate at the population-averaged level during adipogenesis. However, differentiating preadipocytes are highly heterogeneous in cellular and lipid droplet (LD) morphologies, and the degree to which individual cells follow populati...
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| Veröffentlicht in: | The Journal of cell biology 2009-11, Vol.187 (3), p.375-384 |
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| creator | Loo, Lit-Hsin Lin, Hai-Jui Singh, Dinesh K Lyons, Kathleen M Altschuler, Steven J Wu, Lani F |
| description | Increases in key components of adipogenesis and lipolysis pathways correlate at the population-averaged level during adipogenesis. However, differentiating preadipocytes are highly heterogeneous in cellular and lipid droplet (LD) morphologies, and the degree to which individual cells follow population-averaged trends is unclear. In this study, we analyze the molecular heterogeneity of differentiating 3T3-L1 preadipocytes using immunofluorescence microscopy. Unexpectedly, we only observe a small percentage of cells with high simultaneous expression of markers for adipogenesis (peroxisome proliferator-activated receptor γ [PPARγ], CCAAT/enhancer-binding protein α, and adiponectin) and lipid accumulation (hormone-sensitive lipase, perilipin A, and LDs). Instead, we identify subpopulations of cells with negatively correlated expressions of these readouts. Acute perturbation of adipocyte differentiation with PPARγ agonists, forskolin, and fatty acids induced subpopulation-specific effects, including redistribution of the percentage of cells in observed subpopulations and differential expression levels of PPARγ. Collectively, our results suggested that heterogeneity observed during 3T3-L1 adipogenesis reflects a dynamic mixture of subpopulations with distinct physiological states. |
| doi_str_mv | 10.1083/jcb.200904140 |
| format | Article |
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However, differentiating preadipocytes are highly heterogeneous in cellular and lipid droplet (LD) morphologies, and the degree to which individual cells follow population-averaged trends is unclear. In this study, we analyze the molecular heterogeneity of differentiating 3T3-L1 preadipocytes using immunofluorescence microscopy. Unexpectedly, we only observe a small percentage of cells with high simultaneous expression of markers for adipogenesis (peroxisome proliferator-activated receptor γ [PPARγ], CCAAT/enhancer-binding protein α, and adiponectin) and lipid accumulation (hormone-sensitive lipase, perilipin A, and LDs). Instead, we identify subpopulations of cells with negatively correlated expressions of these readouts. Acute perturbation of adipocyte differentiation with PPARγ agonists, forskolin, and fatty acids induced subpopulation-specific effects, including redistribution of the percentage of cells in observed subpopulations and differential expression levels of PPARγ. Collectively, our results suggested that heterogeneity observed during 3T3-L1 adipogenesis reflects a dynamic mixture of subpopulations with distinct physiological states.</description><identifier>ISSN: 0021-9525</identifier><identifier>EISSN: 1540-8140</identifier><identifier>DOI: 10.1083/jcb.200904140</identifier><identifier>PMID: 19948481</identifier><identifier>CODEN: JCLBA3</identifier><language>eng</language><publisher>United States: The Rockefeller University Press</publisher><subject>3T3 L1 cells ; Adipocytes ; Adipocytes - cytology ; Adipocytes - drug effects ; Adipogenesis - physiology ; Adiponectin - metabolism ; Agonists ; Animals ; Antibodies ; Biochemistry ; Body fat ; CCAAT-Binding Factor - metabolism ; Cell Differentiation - drug effects ; Cellular differentiation ; Colforsin - pharmacology ; Fatty acids ; Fatty Acids - pharmacology ; Genetic Markers ; Imaging ; Lipid Metabolism ; Lipids ; Lipogenesis ; Mice ; Microscopy ; Microscopy, Fluorescence ; NIH 3T3 Cells ; Phenotypes ; PPAR gamma - agonists ; PPAR gamma - metabolism ; Proteins ; Studies ; Time Factors</subject><ispartof>The Journal of cell biology, 2009-11, Vol.187 (3), p.375-384</ispartof><rights>Copyright Rockefeller University Press Nov 2, 2009</rights><rights>2009 Loo et al. 2009</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c525t-890ab57f7ddfb889a9bdb2b273571083c1ec49b615e0f3dc6fb3f4d2f53bba1b3</citedby><cites>FETCH-LOGICAL-c525t-890ab57f7ddfb889a9bdb2b273571083c1ec49b615e0f3dc6fb3f4d2f53bba1b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,776,780,881,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19948481$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Loo, Lit-Hsin</creatorcontrib><creatorcontrib>Lin, Hai-Jui</creatorcontrib><creatorcontrib>Singh, Dinesh K</creatorcontrib><creatorcontrib>Lyons, Kathleen M</creatorcontrib><creatorcontrib>Altschuler, Steven J</creatorcontrib><creatorcontrib>Wu, Lani F</creatorcontrib><title>Heterogeneity in the physiological states and pharmacological responses of differentiating 3T3-L1 preadipocytes</title><title>The Journal of cell biology</title><addtitle>J Cell Biol</addtitle><description>Increases in key components of adipogenesis and lipolysis pathways correlate at the population-averaged level during adipogenesis. However, differentiating preadipocytes are highly heterogeneous in cellular and lipid droplet (LD) morphologies, and the degree to which individual cells follow population-averaged trends is unclear. In this study, we analyze the molecular heterogeneity of differentiating 3T3-L1 preadipocytes using immunofluorescence microscopy. Unexpectedly, we only observe a small percentage of cells with high simultaneous expression of markers for adipogenesis (peroxisome proliferator-activated receptor γ [PPARγ], CCAAT/enhancer-binding protein α, and adiponectin) and lipid accumulation (hormone-sensitive lipase, perilipin A, and LDs). Instead, we identify subpopulations of cells with negatively correlated expressions of these readouts. Acute perturbation of adipocyte differentiation with PPARγ agonists, forskolin, and fatty acids induced subpopulation-specific effects, including redistribution of the percentage of cells in observed subpopulations and differential expression levels of PPARγ. Collectively, our results suggested that heterogeneity observed during 3T3-L1 adipogenesis reflects a dynamic mixture of subpopulations with distinct physiological states.</description><subject>3T3 L1 cells</subject><subject>Adipocytes</subject><subject>Adipocytes - cytology</subject><subject>Adipocytes - drug effects</subject><subject>Adipogenesis - physiology</subject><subject>Adiponectin - metabolism</subject><subject>Agonists</subject><subject>Animals</subject><subject>Antibodies</subject><subject>Biochemistry</subject><subject>Body fat</subject><subject>CCAAT-Binding Factor - metabolism</subject><subject>Cell Differentiation - drug effects</subject><subject>Cellular differentiation</subject><subject>Colforsin - pharmacology</subject><subject>Fatty acids</subject><subject>Fatty Acids - pharmacology</subject><subject>Genetic Markers</subject><subject>Imaging</subject><subject>Lipid Metabolism</subject><subject>Lipids</subject><subject>Lipogenesis</subject><subject>Mice</subject><subject>Microscopy</subject><subject>Microscopy, Fluorescence</subject><subject>NIH 3T3 Cells</subject><subject>Phenotypes</subject><subject>PPAR gamma - agonists</subject><subject>PPAR gamma - metabolism</subject><subject>Proteins</subject><subject>Studies</subject><subject>Time Factors</subject><issn>0021-9525</issn><issn>1540-8140</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpdkU1vEzEURS1ERUNhyRIYsWE15fljMvamEqqAVorUBe3asj32xNHEHmwHKf8eR4kCdGVL57yr-_QQeofhGgOnXzZGXxMAAQwzeIEWuGPQ8vp_iRYABLeiI90lep3zBgBYz-grdImFYJxxvEDxzhab4miD9WXf-NCUtW3m9T77OMXRGzU1uahic6PCUIFKW2XOKNk8x5Arja4ZvHM22VC8Kj6MDX2k7Qo3c7Jq8HM0-5ryBl04NWX79vReoafv3x5v79rVw4_726-r1tS6peUClO561w-D05wLJfSgiSY97frD2gZbw4Re4s6Co4NZOk0dG4jrqNYKa3qFbo65805v7WBqq6QmOSe_VWkvo_LyfxL8Wo7xtyR9LwhjNeDzKSDFXzubi9z6bOw0qWDjLsue0iUwwWg1Pz0zN3GXQt1OEtxjApQepPYomRRzTtadq2CQh41kPaQ8H7L6H_7t_9c-Xa4K74_CJpeYzryO8hpAKv945E5Fqcbks3z6SQBTwEvBKeX0Dz6hrxk</recordid><startdate>20091102</startdate><enddate>20091102</enddate><creator>Loo, Lit-Hsin</creator><creator>Lin, Hai-Jui</creator><creator>Singh, Dinesh K</creator><creator>Lyons, Kathleen M</creator><creator>Altschuler, Steven J</creator><creator>Wu, Lani F</creator><general>The Rockefeller University Press</general><general>Rockefeller University Press</general><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20091102</creationdate><title>Heterogeneity in the physiological states and pharmacological responses of differentiating 3T3-L1 preadipocytes</title><author>Loo, Lit-Hsin ; Lin, Hai-Jui ; Singh, Dinesh K ; Lyons, Kathleen M ; Altschuler, Steven J ; Wu, Lani F</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c525t-890ab57f7ddfb889a9bdb2b273571083c1ec49b615e0f3dc6fb3f4d2f53bba1b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>3T3 L1 cells</topic><topic>Adipocytes</topic><topic>Adipocytes - cytology</topic><topic>Adipocytes - drug effects</topic><topic>Adipogenesis - physiology</topic><topic>Adiponectin - metabolism</topic><topic>Agonists</topic><topic>Animals</topic><topic>Antibodies</topic><topic>Biochemistry</topic><topic>Body fat</topic><topic>CCAAT-Binding Factor - metabolism</topic><topic>Cell Differentiation - drug effects</topic><topic>Cellular differentiation</topic><topic>Colforsin - pharmacology</topic><topic>Fatty acids</topic><topic>Fatty Acids - pharmacology</topic><topic>Genetic Markers</topic><topic>Imaging</topic><topic>Lipid Metabolism</topic><topic>Lipids</topic><topic>Lipogenesis</topic><topic>Mice</topic><topic>Microscopy</topic><topic>Microscopy, Fluorescence</topic><topic>NIH 3T3 Cells</topic><topic>Phenotypes</topic><topic>PPAR gamma - agonists</topic><topic>PPAR gamma - metabolism</topic><topic>Proteins</topic><topic>Studies</topic><topic>Time Factors</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Loo, Lit-Hsin</creatorcontrib><creatorcontrib>Lin, Hai-Jui</creatorcontrib><creatorcontrib>Singh, Dinesh K</creatorcontrib><creatorcontrib>Lyons, Kathleen M</creatorcontrib><creatorcontrib>Altschuler, Steven J</creatorcontrib><creatorcontrib>Wu, Lani F</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>The Journal of cell biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Loo, Lit-Hsin</au><au>Lin, Hai-Jui</au><au>Singh, Dinesh K</au><au>Lyons, Kathleen M</au><au>Altschuler, Steven J</au><au>Wu, Lani F</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Heterogeneity in the physiological states and pharmacological responses of differentiating 3T3-L1 preadipocytes</atitle><jtitle>The Journal of cell biology</jtitle><addtitle>J Cell Biol</addtitle><date>2009-11-02</date><risdate>2009</risdate><volume>187</volume><issue>3</issue><spage>375</spage><epage>384</epage><pages>375-384</pages><issn>0021-9525</issn><eissn>1540-8140</eissn><coden>JCLBA3</coden><abstract>Increases in key components of adipogenesis and lipolysis pathways correlate at the population-averaged level during adipogenesis. However, differentiating preadipocytes are highly heterogeneous in cellular and lipid droplet (LD) morphologies, and the degree to which individual cells follow population-averaged trends is unclear. In this study, we analyze the molecular heterogeneity of differentiating 3T3-L1 preadipocytes using immunofluorescence microscopy. Unexpectedly, we only observe a small percentage of cells with high simultaneous expression of markers for adipogenesis (peroxisome proliferator-activated receptor γ [PPARγ], CCAAT/enhancer-binding protein α, and adiponectin) and lipid accumulation (hormone-sensitive lipase, perilipin A, and LDs). Instead, we identify subpopulations of cells with negatively correlated expressions of these readouts. Acute perturbation of adipocyte differentiation with PPARγ agonists, forskolin, and fatty acids induced subpopulation-specific effects, including redistribution of the percentage of cells in observed subpopulations and differential expression levels of PPARγ. 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| subjects | 3T3 L1 cells Adipocytes Adipocytes - cytology Adipocytes - drug effects Adipogenesis - physiology Adiponectin - metabolism Agonists Animals Antibodies Biochemistry Body fat CCAAT-Binding Factor - metabolism Cell Differentiation - drug effects Cellular differentiation Colforsin - pharmacology Fatty acids Fatty Acids - pharmacology Genetic Markers Imaging Lipid Metabolism Lipids Lipogenesis Mice Microscopy Microscopy, Fluorescence NIH 3T3 Cells Phenotypes PPAR gamma - agonists PPAR gamma - metabolism Proteins Studies Time Factors |
| title | Heterogeneity in the physiological states and pharmacological responses of differentiating 3T3-L1 preadipocytes |
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