TY - JOUR
T1 - Decreased caveolae in AGPAT2 lacking adipocytes is independent of changes in cholesterol or sphingolipid levels
T2 - A whole cell and plasma membrane lipidomic analysis of adipogenesis
AU - González-Hódar, Lila
AU - McDonald, Jeffrey G.
AU - Vale, Goncalo
AU - Thompson, Bonne M.
AU - Figueroa, Ana María
AU - Tapia, Pablo J.
AU - Robledo, Fermín
AU - Agarwal, Anil K.
AU - Garg, Abhimanyu
AU - Horton, Jay D.
AU - Cortés, Víctor
N1 - Funding Information:
FONDECYT ( 1181214 ) and National Institutes of Health ( HL-20948 ) provided support to V.C and J.D.H., respectively. We thank Judy Sanchez and Manuel González for their technical support with mouse colonies, Bharat Jaishy for his technical support and Mary Tunison for her scientific editing of the text. L.G. was supported by CONICYT Doctoral Fellowship program grant 21171491.
Funding Information:
FONDECYT (1181214) and National Institutes of Health (HL-20948) provided support to V.C and J.D.H. respectively. We thank Judy Sanchez and Manuel Gonz?lez for their technical support with mouse colonies, Bharat Jaishy for his technical support and Mary Tunison for her scientific editing of the text. L.G. was supported by CONICYT Doctoral Fellowship program grant 21171491.
Publisher Copyright:
© 2021 Elsevier B.V.
PY - 2021/9/1
Y1 - 2021/9/1
N2 - Background: Adipocytes from lipodystrophic Agpat2−/− mice have impaired adipogenesis and fewer caveolae. Herein, we examined whether these defects are associated with changes in lipid composition or abnormal levels of caveolae-associated proteins. Lipidome changes were quantified in differentiated Agpat2−/− adipocytes to identify lipids with potential adipogenic roles. Methods: Agpat2−/− and wild type brown preadipocytes were differentiated in vitro. Plasma membrane was purified by ultracentrifugation. Number of caveolae and caveolae-associated proteins, as well as sterol, sphingolipid, and phospholipid lipidome were determined across differentiation. Results: Differentiated Agpat2−/− adipocytes had decreased caveolae number but conserved insulin signaling. Caveolin-1 and cavin-1 levels were equivalent between Agpat2−/− and wild type adipocytes. No differences in PM cholesterol and sphingolipids abundance were detected between genotypes. Levels of phosphatidylserine at day 10 of differentiation were increased in Agpat2−/− adipocytes. Wild type adipocytes had increased whole cell triglyceride, diacylglycerol, phosphatidylglycerol, phosphatidic acid, lysophosphatidylcholine, lysophosphatidylethanolamine, and trihexosyl ceramide, and decreased 24,25-dihydrolanosterol and sitosterol, as a result of adipogenic differentiation. By contrast, adipogenesis did not modify whole cell neutral lipids but increased lysophosphatidylcholine, sphingomyelin, and trihexosyl ceramide levels in Agpat2−/− adipocytes. Unexpectedly, adipogenesis decreased PM levels of main phospholipids in both genotypes. Conclusion: In Agpat2−/− adipocytes, decreased caveolae is not associated with changes in PM cholesterol nor sphingolipid levels; however, increased PM phosphatidylserine content may be implicated. Abnormal lipid composition is associated with the adipogenic abnormalities of Agpat2 −/− adipocytes but does not prevent insulin signaling.
AB - Background: Adipocytes from lipodystrophic Agpat2−/− mice have impaired adipogenesis and fewer caveolae. Herein, we examined whether these defects are associated with changes in lipid composition or abnormal levels of caveolae-associated proteins. Lipidome changes were quantified in differentiated Agpat2−/− adipocytes to identify lipids with potential adipogenic roles. Methods: Agpat2−/− and wild type brown preadipocytes were differentiated in vitro. Plasma membrane was purified by ultracentrifugation. Number of caveolae and caveolae-associated proteins, as well as sterol, sphingolipid, and phospholipid lipidome were determined across differentiation. Results: Differentiated Agpat2−/− adipocytes had decreased caveolae number but conserved insulin signaling. Caveolin-1 and cavin-1 levels were equivalent between Agpat2−/− and wild type adipocytes. No differences in PM cholesterol and sphingolipids abundance were detected between genotypes. Levels of phosphatidylserine at day 10 of differentiation were increased in Agpat2−/− adipocytes. Wild type adipocytes had increased whole cell triglyceride, diacylglycerol, phosphatidylglycerol, phosphatidic acid, lysophosphatidylcholine, lysophosphatidylethanolamine, and trihexosyl ceramide, and decreased 24,25-dihydrolanosterol and sitosterol, as a result of adipogenic differentiation. By contrast, adipogenesis did not modify whole cell neutral lipids but increased lysophosphatidylcholine, sphingomyelin, and trihexosyl ceramide levels in Agpat2−/− adipocytes. Unexpectedly, adipogenesis decreased PM levels of main phospholipids in both genotypes. Conclusion: In Agpat2−/− adipocytes, decreased caveolae is not associated with changes in PM cholesterol nor sphingolipid levels; however, increased PM phosphatidylserine content may be implicated. Abnormal lipid composition is associated with the adipogenic abnormalities of Agpat2 −/− adipocytes but does not prevent insulin signaling.
KW - Adipogenesis
KW - Caveolae
KW - Lipids
KW - Lipodystrophy
KW - Plasma membrane
KW - agpat2
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UR - http://www.scopus.com/inward/citedby.url?scp=85106916576&partnerID=8YFLogxK
U2 - 10.1016/j.bbadis.2021.166167
DO - 10.1016/j.bbadis.2021.166167
M3 - Article
C2 - 33989739
AN - SCOPUS:85106916576
SN - 0925-4439
VL - 1867
JO - Biochimica et Biophysica Acta - Molecular Basis of Disease
JF - Biochimica et Biophysica Acta - Molecular Basis of Disease
IS - 9
M1 - 166167
ER -