Abstract
Fetal rat brown adipocytes show high-affinity binding sites for both insulin-like growth factor I (IGF-I) and insulin. Cell culture for 24 h in the presence of IGF-I or insulin, independently, up-regulated the mRNA expression of adipogenic-related genes, such as fatty acid synthase (FAS), glycerol-3-phosphate de-hydrogenase and insulin-regulated glucose transporter Glut4, and down-regulated the expression of phosphoenolpyruvate carboxykinase mRNA in a dose-dependent manner. Moreover, both IGF-I and insulin increased the FAS gene transcription rate at 2 h, producing a time-dependent accumulation of FAS mRNA. Furthermore IGF-I or insulin increased glucose uptake and lipid content throughout the 24 h culture period. Our results suggest that both IGF-I and insulin are major signals involved in initiating and/or maintaining the expression of adipogenic-related genes in fetal rat brown adipocytes.
Full Text
The Full Text of this article is available as a PDF (409.2 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Amasino R. M. Acceleration of nucleic acid hybridization rate by polyethylene glycol. Anal Biochem. 1986 Feb 1;152(2):304–307. doi: 10.1016/0003-2697(86)90413-6. [DOI] [PubMed] [Google Scholar]
- Amy C. M., Witkowski A., Naggert J., Williams B., Randhawa Z., Smith S. Molecular cloning and sequencing of cDNAs encoding the entire rat fatty acid synthase. Proc Natl Acad Sci U S A. 1989 May;86(9):3114–3118. doi: 10.1073/pnas.86.9.3114. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Beale E. G., Tishler E. J. Expression and regulation of cytosolic phosphoenolpyruvate carboxykinase in 3T3-L1 adipocytes. Biochem Biophys Res Commun. 1992 Dec 15;189(2):925–930. doi: 10.1016/0006-291x(92)92292-6. [DOI] [PubMed] [Google Scholar]
- Birnbaum M. J. Identification of a novel gene encoding an insulin-responsive glucose transporter protein. Cell. 1989 Apr 21;57(2):305–315. doi: 10.1016/0092-8674(89)90968-9. [DOI] [PubMed] [Google Scholar]
- Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
- Cleveland D. W., Lopata M. A., MacDonald R. J., Cowan N. J., Rutter W. J., Kirschner M. W. Number and evolutionary conservation of alpha- and beta-tubulin and cytoplasmic beta- and gamma-actin genes using specific cloned cDNA probes. Cell. 1980 May;20(1):95–105. doi: 10.1016/0092-8674(80)90238-x. [DOI] [PubMed] [Google Scholar]
- Foufelle F., Gouhot B., Perdereau D., Girard J., Ferre P. Regulation of lipogenic enzyme and phosphoenolpyruvate carboxykinase gene expression in cultured white adipose tissue. Glucose and insulin effects are antagonized by cAMP. Eur J Biochem. 1994 Aug 1;223(3):893–900. doi: 10.1111/j.1432-1033.1994.tb19066.x. [DOI] [PubMed] [Google Scholar]
- Foufelle F., Gouhot B., Pégorier J. P., Perdereau D., Girard J., Ferré P. Glucose stimulation of lipogenic enzyme gene expression in cultured white adipose tissue. A role for glucose 6-phosphate. J Biol Chem. 1992 Oct 15;267(29):20543–20546. [PubMed] [Google Scholar]
- Greenspan P., Mayer E. P., Fowler S. D. Nile red: a selective fluorescent stain for intracellular lipid droplets. J Cell Biol. 1985 Mar;100(3):965–973. doi: 10.1083/jcb.100.3.965. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ireland R. C., Kotarski M. A., Johnston L. A., Stadler U., Birkenmeier E., Kozak L. P. Primary structure of the mouse glycerol-3-phosphate dehydrogenase gene. J Biol Chem. 1986 Sep 5;261(25):11779–11785. [PubMed] [Google Scholar]
- Lorenzo M., Fabregat I., Benito M. Hormonal regulation of malic enzyme expression in primary cultures of foetal brown adipocytes. Biochem Biophys Res Commun. 1989 Aug 30;163(1):341–347. doi: 10.1016/0006-291x(89)92141-4. [DOI] [PubMed] [Google Scholar]
- Lorenzo M., Roncero C., Fabregat I., Benito M. Hormonal regulation of rat foetal lipogenesis in brown-adipocyte primary cultures. Biochem J. 1988 Apr 15;251(2):617–620. doi: 10.1042/bj2510617. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lorenzo M., Valverde A. M., Teruel T., Benito M. IGF-I is a mitogen involved in differentiation-related gene expression in fetal rat brown adipocytes. J Cell Biol. 1993 Dec;123(6 Pt 1):1567–1575. doi: 10.1083/jcb.123.6.1567. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Misra S., Sakamoto K., Moustaïd N., Sul H. S. Localization of sequences for the basal and insulin-like growth factor-I inducible activity of the fatty acid synthase promoter in 3T3-L1 fibroblasts. Biochem J. 1994 Mar 15;298(Pt 3):575–578. doi: 10.1042/bj2980575. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moustaïd N., Beyer R. S., Sul H. S. Identification of an insulin response element in the fatty acid synthase promoter. J Biol Chem. 1994 Feb 25;269(8):5629–5634. [PubMed] [Google Scholar]
- Moustaïd N., Sul H. S. Regulation of expression of the fatty acid synthase gene in 3T3-L1 cells by differentiation and triiodothyronine. J Biol Chem. 1991 Oct 5;266(28):18550–18554. [PubMed] [Google Scholar]
- Prip-Buus C., Perdereau D., Foufelle F., Maury J., Ferre P., Girard J. Induction of fatty-acid-synthase gene expression by glucose in primary culture of rat hepatocytes. Dependency upon glucokinase activity. Eur J Biochem. 1995 May 15;230(1):309–315. doi: 10.1111/j.1432-1033.1995.0309i.x. [DOI] [PubMed] [Google Scholar]
- Rice K. M., Garner C. W. Correlation of the insulin receptor substrate-1 with insulin-responsive deoxyglucose transport in 3T3-L1 adipocytes. Biochem Biophys Res Commun. 1994 Jan 28;198(2):523–530. doi: 10.1006/bbrc.1994.1077. [DOI] [PubMed] [Google Scholar]
- Roberts S., Bentley D. L. Distinct modes of transcription read through or terminate at the c-myc attenuator. EMBO J. 1992 Mar;11(3):1085–1093. doi: 10.1002/j.1460-2075.1992.tb05147.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rubin C. S., Hirsch A., Fung C., Rosen O. M. Development of hormone receptors and hormonal responsiveness in vitro. Insulin receptors and insulin sensitivity in the preadipocyte and adipocyte forms of 3T3-L1 cells. J Biol Chem. 1978 Oct 25;253(20):7570–7578. [PubMed] [Google Scholar]
- Santalucía T., Camps M., Castelló A., Muñoz P., Nuel A., Testar X., Palacin M., Zorzano A. Developmental regulation of GLUT-1 (erythroid/Hep G2) and GLUT-4 (muscle/fat) glucose transporter expression in rat heart, skeletal muscle, and brown adipose tissue. Endocrinology. 1992 Feb;130(2):837–846. doi: 10.1210/endo.130.2.1370797. [DOI] [PubMed] [Google Scholar]
- Shima A., Shinohara Y., Doi K., Terada H. Normal differentiation of rat brown adipocytes in primary culture judged by their expressions of uncoupling protein and the physiological isoform of glucose transporter. Biochim Biophys Acta. 1994 Aug 11;1223(1):1–8. doi: 10.1016/0167-4889(94)90066-3. [DOI] [PubMed] [Google Scholar]
- Sivitz W. I., DeSautel S. L., Kayano T., Bell G. I., Pessin J. E. Regulation of glucose transporter messenger RNA in insulin-deficient states. Nature. 1989 Jul 6;340(6228):72–74. doi: 10.1038/340072a0. [DOI] [PubMed] [Google Scholar]
- Smas C. M., Sul H. S. Control of adipocyte differentiation. Biochem J. 1995 Aug 1;309(Pt 3):697–710. doi: 10.1042/bj3090697. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smyth M. J., Sparks R. L., Wharton W. Proadipocyte cell lines: models of cellular proliferation and differentiation. J Cell Sci. 1993 Sep;106(Pt 1):1–9. doi: 10.1242/jcs.106.1.1. [DOI] [PubMed] [Google Scholar]
- Smyth M. J., Wharton W. Differentiation of A31T6 proadipocytes to adipocytes: a flow cytometric analysis. Exp Cell Res. 1992 Mar;199(1):29–38. doi: 10.1016/0014-4827(92)90458-k. [DOI] [PubMed] [Google Scholar]
- Stephens J. M., Pilch P. F. The metabolic regulation and vesicular transport of GLUT4, the major insulin-responsive glucose transporter. Endocr Rev. 1995 Aug;16(4):529–546. doi: 10.1210/edrv-16-4-529. [DOI] [PubMed] [Google Scholar]
- Teruel T., Valverde A. M., Alvarez A., Benito M., Lorenzo M. Differentiation of rat brown adipocytes during late foetal development: role of insulin-like growth factor I. Biochem J. 1995 Sep 15;310(Pt 3):771–776. doi: 10.1042/bj3100771. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Teruel T., Valverde A. M., Benito M., Lorenzo M. Transforming growth factor beta 1 induces differentiation-specific gene expression in fetal rat brown adipocytes. FEBS Lett. 1995 May 8;364(2):193–197. doi: 10.1016/0014-5793(95)00385-m. [DOI] [PubMed] [Google Scholar]
- Valverde A. M., Benito M., Lorenzo M. Hormonal regulation of malic enzyme and glucose-6-phosphate-dehydrogenase expression in fetal brown-adipocyte primary cultures under non-proliferative conditions. Eur J Biochem. 1992 Jan 15;203(1-2):313–319. doi: 10.1111/j.1432-1033.1992.tb19861.x. [DOI] [PubMed] [Google Scholar]
- Valverde A. M., Benito M., Lorenzo M. Proliferation of fetal brown adipocyte primary cultures: relationship with the genetic expression of glucose 6 phosphate dehydrogenase. Exp Cell Res. 1991 Jun;194(2):232–237. doi: 10.1016/0014-4827(91)90359-3. [DOI] [PubMed] [Google Scholar]
- Valverde A. M., Lorenzo M., Teruel T., Benito M. cAMP inhibits IGF-I-induced mitogenesis in fetal rat brown adipocytes: role of p21 ras. Exp Cell Res. 1995 May;218(1):305–309. doi: 10.1006/excr.1995.1159. [DOI] [PubMed] [Google Scholar]
- Yoo-Warren H., Monahan J. E., Short J., Short H., Bruzel A., Wynshaw-Boris A., Meisner H. M., Samols D., Hanson R. W. Isolation and characterization of the gene coding for cytosolic phosphoenolpyruvate carboxykinase (GTP) from the rat. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3656–3660. doi: 10.1073/pnas.80.12.3656. [DOI] [PMC free article] [PubMed] [Google Scholar]