Follistatin rescues the inhibitory effect of activin A on the differentiation of bovine preadipocyte.

We investigated the effect of activin A and follistatin on the differentiation of bovine preadipocytes. Stromal-vascular (SV) cells containing preadipocytes were prepared from perirenal adipose tissue of approximately 30-month-old Japanese Black steers. After confluence, differentiation was induced by 1-methyl-3-isobutyl-xanthine, dexamethasone, insulin and troglitasone for 2 days, and then subsequently cultured for 6 days. The cells were treated with activin A during the induction of differentiation (the early phase of differentiation) or throughout the differentiation period. We measured the terminal differentiation markers such as glycerol-3-phosphate dehydrogenase (GPDH) activity, lipid accumulation, and the expression of adipocyte fatty acid-binding protein mRNA at the end of cultures. Activin A suppressed the induction of all differentiation markers regardless of the duration of treatment. The treatment with activin A also reduced the expression of peroxisome proliferator-activated receptor (PPAR) gamma and CCAAT/enhancer binding protein (C/EBP) alpha mRNAs without affecting the expression of C/EBPbeta mRNA. We also observed that follistatin completely rescued the inhibitory effect of activin A on bovine preadipocyte differentiation. Furthermore, the higher doses of follistatin increased GPDH activity even in the presence of activin A compared with the cells treated with neither activin A nor follistatin. Additionally, the SV cells expressed activin A and myostatin mRNAs. These results suggest that activin A inhibits bovine preadiopocyte differentiation via affecting transcriptional cascade upstream of PPARgamma and C/EBPalpha expressions, and that follistatin suppresses the inhibitory effect of activin A on bovine preadipocyte differentiation. Endogenous activin A and/or myostatin possibly inhibit the differentiation of bovine preadipocytes.

Nitric oxide suppresses preadipocyte differentiation in 3T3-L1 culture.

Nitric oxide (NO) is an important chemical messenger controlling many physiological functions, involving cell proliferation, and differentiation. The purpose of this study was to investigate the effect of NO on adipocyte differentiation using a murine preadipocyte cell line, 3T3-L1. The treatment with a NO donor, 1-hydroxy-2-oxo-3,3-bis(2-aminoethyl)-1-triazene (NOC18), reduced some markers of adipocyte differentiation such as glycerol-3-phosphate dehydrogenase activity, and intracellular lipid accumulation. To examine whether these effects of NOC18 on adipocyte differentiation markers are due to its cytotoxity, lactate dehydrogenase (LDH) release from the cells were measured. NOC18 did not affect LDH release into the culture medium. Thus, the suppressive actions of NO donor were unlikely to result from its cytotoxicity. Peroxisome proliferator-activated receptor (PPAR) gamma is a critical transcription factor for adipocyte differentiation and adipocyte fatty acid binding protein (aP2) gene is one of its targets. Protein expression of PPARgamma was not diminished by NOC18 treatment, although mRNA expression of aP2 was reduced. Electrophoretic mobility shift assay showed that NOC18 interfered with the DNA binding activity of PPARgamma. Therefore, the present experiment suggest that NO suppresses adipocyte differentiation through suppressing the transcriptional activity of PPARgamma, without suppressing its expression level.