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Bone morphogenetic protein-3b (BMP-3b) is expressed in adipocytes and inhibits adipogenesis as a unique complex.

ABSTRACT: BACKGROUND: Bone morphogenetic protein-3b (BMP-3b) is a member of the transforming growth factor-? (TGF-?) superfamily. BMP-3b regulates osteogenesis and has critical roles in developing embryos. BMP-3b is expressed not only in the bone and developing embryos but also in adipose tissues. However, the functions of BMP-3b in adipose tissue are still unknown. METHODS: BMP-3b expression was quantified in various adipose tissues and in the adipose-derived stromal-vascular fraction (SVF) and mature adipocyte fraction (AD.F) of mice. We also used 3T3-L1 preadipocytes to analyze the expression, function and molecular forms of BMP-3b. In order to determine the effects of BMP-3b on the adipogenesis of 3T3-L1 cells, BMP-3b siRNA-mediated knockdown and gene overexpression studies were performed, and a conditioned medium (CM) containing the BMP-3b protein was added to 3T3-L1 cell cultures. Adipocyte differentiation was evaluated by measuring the expression of adipogenic markers or by Oil Red O staining. The molecular form of BMP-3b that was secreted from the 3T3-L1 cells was analyzed by western blotting. RESULTS: BMP-3b is expressed in all adipose tissues and is expressed at higher levels in preadipocytes than in mature adipocytes. In mesenteric adipose tissue, BMP-3b expression was increased in diet-induced obesity (DIO) mice as compared with that in control mice. BMP-3b was also expressed highly in 3T3-L1 cells. We showed that siRNA-mediated knockdown of endogenous BMP-3b expression in 3T3-L1 cells enhanced adipogenesis. Conversely, overexpressing BMP-3b inhibited adipocyte differentiation. We also showed that addition of CM containing the BMP-3b protein inhibited the differentiation of 3T3-L1 cells, and that this inhibitory effect was abolished by removing BMP-3b with an anti-BMP-3b antibody. Furthermore, BMP-3b was secreted from adipocytes as a unique non-covalent complex. CONCLUSION: These data suggest that BMP-3b is secreted from adipocytes and is involved in adipocyte differentiation.

SUBMITTER: Hino J

PROVIDER: S-EPMC3348488 | biostudies-other | 2012 May

REPOSITORIES: biostudies-other

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Bone morphogenetic protein-3b (BMP-3b) is expressed in adipocytes and inhibits adipogenesis as a unique complex.

Hino J J Miyazawa T T Miyazato M M Kangawa K K

International journal of obesity (2005) 20110628 5

<h4>Background</h4>Bone morphogenetic protein-3b (BMP-3b) is a member of the transforming growth factor-β (TGF-β) superfamily. BMP-3b regulates osteogenesis and has critical roles in developing embryos. BMP-3b is expressed not only in the bone and developing embryos but also in adipose tissues. However, the functions of BMP-3b in adipose tissue are still unknown.<h4>Methods</h4>BMP-3b expression was quantified in various adipose tissues and in the adipose-derived stromal-vascular fraction (SVF) ...[more]

PMID: 21712809

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Project description:BACKGROUND:Bone morphogenetic proteins (BMPs) regulate adipogenesis but it is not clear whether they influence regional adipose tissue (AT) development in humans. OBJECTIVE:To characterise BMP2 expression, BMP2-SMAD1/5/8 signalling, and BMP2's potential effect on proliferation and adipogenesis in human subcutaneous abdominal and gluteal AT and its constituent preadipocytes. METHODS:BMP2 expression was measured in whole AT and immortalised preadipocytes via qPCR and Western blot; secreted/circulating BMP2 was measured by ELISA. The effect of BMP2 on preadipocyte proliferation was evaluated using a fluorescent assay. BMP2's effect on adipogenesis in immortalised preadipocytes was determined via qPCR of adipogenic markers and cellular triacylglycerol (TAG) accumulation. BMP2-SMAD1/5/8 signalling was assessed in immortalised preadipocytes via Western blot and qPCR of ID1 expression. RESULTS:BMP2 was expressed and released by abdominal and gluteal AT and preadipocytes. Exogenous BMP2 dose dependently promoted adipogenesis in abdominal preadipocytes only; 50?ng/ml BMP2 increased PPARG2 expression (10-fold compared to vehicle, p?<?0.001) and TAG accumulation (3-fold compared to vehicle; p?<?0.001). BMP2 stimulated SMAD1/5/8 phosphorylation and ID1 expression in abdominal and gluteal preadipocytes but this was blocked by 500?nM K02288, a type 1 BMP receptor inhibitor (p?<?0.001). Co-administration of 500?nM K02288 also inhibited the pro-adipogenic effect of 50?ng/ml BMP2 in abdominal cells; >90% inhibition of TAG accumulation (p?<?0.001) and ~50% inhibition of PPARG2 expression (p?<?0.001). The endogenous iron regulator erythroferrone reduced BMP2-SMAD1/5/8 signalling by ~30% specifically in subcutaneous abdominal preadipocytes (p?<?0.01), suggesting it plays a role in restricting the expansion of the body's largest AT depot during energy deficiency. Additionally, a waist-hip ratio-increasing common polymorphism near BMP2 is an eQTL associated with ~15% lower BMP2 expression in abdominal and gluteal AT (p?<?0.05) as well as altered adipocyte size in male abdominal AT (p?<?0.05). CONCLUSIONS:These data implicate BMP2-SMAD1/5/8 signalling in depot-specific preadipocyte development and abdominal AT expansion in humans.

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Dataset Information

Bone morphogenetic protein-3b (BMP-3b) is expressed in adipocytes and inhibits adipogenesis as a unique complex.

Publications

Bone morphogenetic protein-3b (BMP-3b) is expressed in adipocytes and inhibits adipogenesis as a unique complex.

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