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Journal of Lipid Research, Vol. 43, 2123-2129, December 2002
Copyright © 2002 by Lipid Research, Inc.
Nitric oxide promotes differentiation of rat white preadipocytes in culture
* Julia McFarlane Diabetes Research Centre, Departments of Medicine, Biochemistry and Molecular Biology, The University of Calgary, Calgary, AB, T2N 4N1
Department of Medicine and the Loeb Health Research Institute, The Ottawa Hospital and the University of Ottawa, Ottawa, ON, K1Y 4E9, Canada
1 To whom correspondence should be addressed. e-mail: dcwlau{at}ucalgary.ca
The putative role of nitric oxide (NO) in modulating adipogenesis was investigated in cultured preadipocytes derived from rat white adipose tissue. The NO releasing reagent, hydroxylamine (HA), and nitric oxide synthase (NOS) substrate L-arginine (Arg) had no influence on cell replication. However, both HA and Arg exhibited significant induction on differentiation, as evidenced by increased lipoprotein lipase (LPL) and glycerol-3-phosphate dehydrogenase (GPDH) activities, as well as accelerated triacylglycerol (TG) accumulation. These observations suggested a positive role of NO in modulating adipogenesis. Preadipocytes were found to produce NO, and a 
50% increase over basal level was observed on the first 2 days of differentiation. Deprivation of endogenous NOS activity by a non-selective NOS inhibitor, NG-monomethyl-L-arginine (NMMA), partially abrogated the differentiation process, implicating a role for endogenous NO to stimulate preadipocyte differentiation. Both NOS isoforms, eNOS and iNOS, were detected in differentiating preadipocytes.
Specific iNOS inhibitors (1400W and aminoguanidine) had little influence on NO production and differentiation, suggesting that eNOS rather than iNOS may be the major isoform involved in modulating adipogenesis.
Abbreviations: ACM, adipocyte-conditioned medium; AG, aminoguanidine; 
MEM, -minimum essential medium; Arg, L-arginine; DD, differentiation day of; eNOS, endothelial nitric oxide synthase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GPDH, glycerol-3-phosphate dehydrogenase; HA, hydroxylamine; iNOS, inducible nitric oxide synthase; LPL, lipoprotein lipase; nNOS, neuronal nitric oxide synthase; NMMA, NG-monomethyl-L-arginine; NO, nitric oxide; PPAR
2, peroxisome-proliferator-activated receptor 2; RT-PCR, reverse transcriptase-polymerase chain reaction; TG, triacylglycerol
Supplementary key words eNOS • iNOS • adipocytes
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