title: Cysteine deprivation prevents induction of peroxisome proliferator-activated receptor gamma-2 and adipose differentiation of 3T3-L1 cells.
abstract: Plasma cysteine is strongly associated with body fat mass in human cohorts and diets low in cysteine prevents fat accumulation in mice. It is unclear if plasma cysteine affects fat development or if fat accumulation raises plasma cysteine. To determine if cysteine affects adipogenesis, we differentiated 3T3-L1 preadipocytes in medium with reduced cysteine. Cells incubated in media with 10-20μM cysteine exhibited reduced capacity to differentiate into triacylglycerol-storing mature adipocytes compared with cells incubated with 50μM cysteine. Low cysteine severely reduced expression of peroxisome proliferator-activated receptor gamma2 (Pparγ2) and its target genes perlipin1 (Plin1) and fatty acid binding protein-4 (Fabp4). Expression of stearoyl-CoA desaturase-1 (Scd1), known to be repressed with cysteine depletion, was also reduced with low cysteine. Medium depletion of the essential amino acids leucine, valine, and isoleucine had only a modest effect on adipocyte specific gene expression and differentiation. Stimulation with the PPARγ agonist BRL-49653 or addition of a hydrogen sulfide donor enhanced differentiation of 3T3-L1 cells cultured in low cysteine. This demonstrates that the ability to induce PPARγ expression is preserved when cells are cultured in low cysteine. It therefore appears that cysteine depletion inhibits adipogenesis by specifically affecting molecular pathways required for induction of PPARγ expression, rather than through a general reduction of global protein synthesis. In conclusion, we show that low extracellular cysteine reduces adipocyte differentiation by interfering with PPARγ2 and PPARγ target gene expression. Our results provide further evidence for the hypothesis that plasma cysteine is a casual determinant for body fat mass.
authors.affiliation: Department of Nutrition, Institute of Basic Medical Sciences, University of Oslo, Norway.
authors.forename: Nadia Nabil
authors.initials: NN
authors.lastname: Haj-Yasein
authors.affiliation: Department of Nutrition, Institute of Basic Medical Sciences, University of Oslo, Norway.
authors.forename: Ole
authors.initials: O
authors.lastname: Berg
authors.affiliation: Department of Pharmacology, University of Oxford, United Kingdom.
authors.forename: Fredrik
authors.initials: F
authors.lastname: Jernerén
authors.affiliation: Department of Nutrition, Institute of Basic Medical Sciences, University of Oslo, Norway; Department of Pharmacology, University of Oxford, United Kingdom.
authors.forename: Helga
authors.initials: H
authors.lastname: Refsum
authors.affiliation: Department of Nutrition, Institute of Basic Medical Sciences, University of Oslo, Norway.
authors.forename: Hilde I
authors.initials: HI
authors.lastname: Nebb
authors.affiliation: Department of Nutrition, Institute of Basic Medical Sciences, University of Oslo, Norway; The Norwegian Transgenic Center, Department of Nutrition, Institute of Basic Medical Sciences, University of Oslo, Norway. Electronic address: k.t.dalen@medisin.uio.no.
authors.forename: Knut Tomas
authors.initials: KT
authors.lastname: Dalen
mesh_terms: D041721:3T3-L1 Cells; D017667:Adipocytes / Q000166:cytology / Q000378:metabolism*; D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D003545:Cysteine / Q000378:metabolism* / Q000494:pharmacology; D050556:Fatty Acid-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000502:physiology; D051379:Mice; D047495:PPAR gamma / Q000235:genetics / Q000378:metabolism*; D000070778:Perilipin-1 / Q000235:genetics / Q000378:metabolism*; D013230:Stearoyl-CoA Desaturase / Q000235:genetics / Q000378:metabolism*
chemical_list: C493411:Fabp4 protein, mouse; D050556:Fatty Acid-Binding Proteins; D047495:PPAR gamma; D000070778:Perilipin-1; C000602979:Plin1 protein, mouse; C490333:Scd1 protein, mouse; D013230:Stearoyl-CoA Desaturase; D003545:Cysteine
keywords: 3T3-L1 cells; Adipocytes; Cysteine; Differentiation; Obesity; Peroxisome proliferator-activated receptor gamma