Sublines of mouse 3T3 cells that accumulate lipid

Green, Howard; Kehinde, Olaniyi

doi:10.1016/0092-8674(74)90126-3

Cited by 912 publications

(465 citation statements)

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“…The murine 3T3-L1 preadipocyte is a model system for the process of adipocyte differentiation (28). Differentiation results in the increased expression of insulin receptors (29), insulinresponsive glucose transporters (30,31), and enzymes required for fatty acid synthesis (32)(33)(34).…”

Section: Resultsmentioning

confidence: 99%

CSF-1 Receptor/Insulin Receptor Chimera Permits CSF-1-dependent Differentiation of 3T3-L1 Preadipocytes

Chaika¹,

Chaika²,

Volle³

et al. 1997

Journal of Biological Chemistry

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A chimeric growth factor receptor (CSF1R/IR) was constructed by splicing cDNA sequences encoding the extracellular ligand binding domain of the human colony stimulating factor-1 (CSF-1) receptor to sequences encoding the transmembrane and cytoplasmic domains of the human insulin receptor. The addition of CSF-1 to cells transfected with the CSF1R/IR chimera cDNA stimulated the tyrosine phosphorylation of a protein that was immunoprecipitated by an antibody directed against the carboxyl terminus of the insulin receptor. Phosphopeptide maps of the 32 P-labeled CSF1R/IR protein revealed the same pattern of phosphorylation observed in 32 P-labeled insulin receptor ␤ subunits. CSF-1 stimulated the tyrosine phosphorylation of insulin receptor substrate-1 (IRS-1) and Shc in cells expressing the CSF1R/IR chimera. Lipid accumulation and the expression of a differentiation-specific marker demonstrated that 3T3-L1 preadipocytes undergo CSF-1-dependent differentiation when transfected with the CSF1R/IR chimera cDNA but not when transfected with the expression vector alone. A 12-amino acid deletion within the juxtamembrane region of the CSF1R/IR (CSF1R/IR⌬960) blocked CSF-1-stimulated phosphorylation of IRS-1 and Shc but did not inhibit CSF-1-mediated differentiation of 3T3-L1 preadipocytes. These observations indicate that adipocyte differentiation can be initiated by intracellular pathways that do not require tyrosine phosphorylation of IRS-1 or Shc.A primary goal in the study of insulin action is the identification of the intracellular pathways that lead ultimately to changes in the rates of growth, development, and metabolism in primary target tissues of insulin (e.g. adipose, muscle, and liver). One strategy for the study of insulin-sensitive intracellular signaling has been to identify structural features within the insulin receptor that are important components of divergent signal transduction pathways. The goal of this strategy has been to create mutations within specific receptor sequences that result in the selective disruption of some insulin-regulated metabolic pathways while leaving others intact. Deletion mutagenesis of the insulin receptor cytoplasmic domain has generated insulin receptors with altered biological properties (1-4). Experiments with these receptor mutants have led to the suggestion that different regions of the insulin receptor cytoplasmic domain may play roles in modulating the distinct biological effects of insulin. Most mutations of the insulin receptor cytoplasmic domain have removed or altered autophosphorylation sites within the cytoplasmic domain (5-8). This approach has furthered the notion that tyrosine phosphorylation and the tyrosine kinase encoded within the receptor ␤ subunit are essential components of normal insulin action. The majority of structure/function analyses of the insulin receptor, however, have been performed in cells expressing low levels of endogenous insulin receptors, e.g. Chinese hamster ovary (CHO) 1 or Rat-1 fibroblasts cell lines (reviewed in Ref. 9). Although su...

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Section: Resultsmentioning

confidence: 99%

CSF-1 Receptor/Insulin Receptor Chimera Permits CSF-1-dependent Differentiation of 3T3-L1 Preadipocytes

Chaika¹,

Chaika²,

Volle³

et al. 1997

Journal of Biological Chemistry

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“…To confirm our findings on the expression of PU.1 in preadipocytes and adipocytes, we used the 3T3-L1 preadipocyte cell line that can be differentiated into adipocytes in vitro (9). Total RNA was collected from 3T3-L1 cells at different time points of adipocyte differentiation.…”

Section: Pu1 Is Expressed In Adipose Tissuementioning

confidence: 99%

Transcription factor PU.1 is expressed in white adipose and inhibits adipocyte differentiation

Wang¹,

Tong²

2008

American Journal of Physiology-Cell Physiology

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PU.1 transcription factor is a critical regulator of hematopoiesis and leukemogenesis. Because PU.1 interacts with transcription factors GATA-2 and C/EBPα, and both are involved in the regulation of adipogenesis, we investigated whether PU.1 plays a role in the regulation of adipocyte differentiation. Our data indicate that PU.1 is expressed in white adipose tissue. PU.1 protein can also be detected in cultured 3T3-L1 adipocytes. Forced expression of PU.1 in 3T3-L1 cells inhibits adipocyte differentiation, whereas deletion of the transactivation domain of PU.1 abolishes this effect. The inhibition of adipocyte differentiation by PU.1 is achieved, at least in part, through repression of the transcriptional activity of C/EBPα and C/EBPβ. Furthermore, GATA-2 and PU.1 have an additive inhibitory effect on C/EBP transactivation and adipogenesis. Finally, the expression of PU.1 is increased in white adipose of obese mice.

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“…Such clonal variants can either arise spontaneously [3T3-L1 from Swiss 3T3 (Green and Kehinde 1974); BALB/c-T-3T3 from BALB/c-3T3 (Diamond et al 1977)] or be generated by treatment of cells with agents that alter DNA methylation {TA1 from C3H10T1/2, Chapman et al 1984). Although these variant cell lines typically show a low frequency of spontaneous differentiation, they can be induced to differentiate at high frequency by treatment with lipogenic hormones [3T3-L1 (Rubin et al 1978)], high concentrations of human plasma [BALB/c-T-3T3 {Scott et al 1982)] or high cell density [TA1 {Chapman et al 1984)].…”

Section: High Stable Expression Of C/ebpa Promotes Adipogenesis M Balmentioning

confidence: 99%

Ectopic expression of the CCAAT/enhancer-binding protein alpha promotes the adipogenic program in a variety of mouse fibroblastic cells.

Freytag

Paielli²,

Gilbert³

1994

Genes Dev.

416

294

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The CCAAT/enhancer-binding protein ot (C/EBPa) has been implicated in the regulation of adipoblast differentiation. In this study we investigate the potential of C/EBPot to promote the adipogenic program in a variety of fibroblastic cells. Transduction of the C/EBPc~ gene into eight mouse fibroblastic cell lines by retroviruses and DNA transfection generates adipocyte colonies at variable frequencies. The most dramatic results are obtained with NIH-3T3 cells, in which the percentage of G418-resistant colonies that exhibit the adipocyte morphology is reproducibly >50% when the C/EBPc~ gene is transduced by retroviruses. The ability to promote the adipogenic program requires the potent transcriptional activation domain of C/EBPa and is not observed with C/EBPI3. Paradoxically, in spite of its antimitogenic effects, clonal cell lines that stably express high amounts of C/EBPa can readily be generated. Stable expression of C/EBPot in BALB/c-3T3 cells dramatically enhances their ability to terminally differentiate into adipocytes. The results demonstrate that C/EBPa can efficiently promote the adipogenic program in a variety of mouse fibroblastic cells, including those that have little or no spontaneous capacity to undergo adipogenesis.

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Sublines of mouse 3T3 cells that accumulate lipid

Cited by 912 publications

References 1 publication

CSF-1 Receptor/Insulin Receptor Chimera Permits CSF-1-dependent Differentiation of 3T3-L1 Preadipocytes

CSF-1 Receptor/Insulin Receptor Chimera Permits CSF-1-dependent Differentiation of 3T3-L1 Preadipocytes

Transcription factor PU.1 is expressed in white adipose and inhibits adipocyte differentiation

Ectopic expression of the CCAAT/enhancer-binding protein alpha promotes the adipogenic program in a variety of mouse fibroblastic cells.

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