scholarly journals Requirement and role of arachidonic acid in the differentiation of pre-adipose cells

1989 ◽  
Vol 257 (2) ◽  
pp. 389-397 ◽  
Author(s):  
D Gaillard ◽  
R Négrel ◽  
M Lagarde ◽  
G Ailhaud
Keyword(s):  
Growth Hormone ◽  
Arachidonic Acid ◽  
Cyclic Amp ◽  
Adipose Conversion ◽  
Inositol Phospholipid ◽  
Glycerol 3 Phosphate Dehydrogenase ◽  
Triacylglycerol Accumulation ◽  
Exogenous Origin ◽  
Inositol Phospholipid Breakdown ◽  
Adipose Cells

The terminal adipose differentiation of Ob1771 cells, characterized by glycerol-3-phosphate dehydrogenase activity and triacylglycerol accumulation, was studied in serum-free hormone-supplemented medium containing growth hormone, tri-iodothyronine, insulin, transferrin and fetuin. Arachidonic acid was able to substitute for a crude adipogenic fraction isolated from fetal bovine serum but not for growth hormone or tri-iodothyronine. Arachidonic acid was also able to increase in a rapid and dramatic manner cyclic AMP production; moreover it was able to amplify the adipose conversion promoted by other agents elevating cyclic AMP concentrations and to induce inositol phospholipid breakdown. Both phorbol 12-myristate 13-acetate, a protein kinase C activator and ionomycin, a Ca2+-mobilizing agent, showed potent synergy with agents elevating cyclic AMP concentrations for the promotion of adipose conversion, whereas 8-bromo cyclic GMP and 4 alpha-phorbol 12,13-dibutyrate were ineffective. The triggering of both the cyclic AMP and inositol phospholipid pathways was accompanied by a single round of cell division, and within a few days all the cells became differentiated. Similar results were obtained, after exposure to arachidonic acid, with preadipose 3T3-F442A cells and with rat adipose precursor cells in primary culture. The availability of arachidonic acid from intracellular stores and/or of exogenous origin should play a major role for the onset of critical mitoses leading to terminal differentiation in pre-adipose cells.

scholarly journals Prostacyclin as a potent effector of adipose-cell differentiation

1989 ◽  
Vol 257 (2) ◽  
pp. 399-405 ◽  
Author(s):  
R Négrel ◽  
D Gaillard ◽  
G Ailhaud
Keyword(s):  
Arachidonic Acid ◽  
Cyclic Amp ◽  
Critical Role ◽  
Terminal Differentiation ◽  
Prostaglandin F2 Alpha ◽  
Adipogenic Effect ◽  
Adipose Cells

The terminal differentiation of Ob1771 pre-adipose cells induced by arachidonic acid in serum-free hormone-supplemented medium containing insulin, transferrin, growth hormone, tri-iodothyronine and fetuin (5F medium) was strongly diminished in the presence of inhibitors of prostaglandin synthesis, namely aspirin or indomethacin. Carbaprostacyclin, a stable analogue of prostacyclin (prostaglandin I2) known to be synthesized by pre-adipocytes and adipocytes, behaved as an efficient activator of cyclic AMP production and was able, when added to 5F medium, to mimic the adipogenic effect of arachidonic acid. Prostaglandins E2, F2 alpha and D2, unable to affect the cyclic AMP production, failed to substitute for carbaprostacyclin. However, prostaglandin F2 alpha, which is another metabolite of arachidonic acid in pre-adipose and adipose cells, able to promote inositol phospholipid breakdown and protein kinase C activation, potentiated the adipogenic effect of carbaprostacyclin. In addition, carbaprostacyclin enhanced both a limited proliferation and terminal differentiation of adipose precursor cells isolated from rodent and human adipose tissues maintained in primary culture. These results demonstrate the critical role of prostacyclin and prostaglandin F2 alpha on adipose conversion in vitro and suggest a paracrine/autocrine role of both prostanoids in the development of adipose tissue in vivo.

scholarly journals Role of spermidine in the expression of late markers of adipose conversion Effects of growth hormone

1986 ◽  
Vol 239 (2) ◽  
pp. 363-370 ◽  
Author(s):  
E Z Amri ◽  
R Barbaras ◽  
A Doglio ◽  
C Dani ◽  
P Grimaldi ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Mrna Level ◽  
Fold Increase ◽  
Intracellular Concentration ◽  
Polyamine Content ◽  
Adipose Conversion ◽  
Highly Correlated ◽  
Glycerol 3 Phosphate Dehydrogenase

Confluent Ob1771 cells treated with an inhibitor of spermidine and spermine synthesis, methylglyoxyal bis(guanylhydrazone), were dependent on putrescine addition for the expression of glycerol-3-phosphate dehydrogenase and acyl-CoA synthetase, which behaved as late markers of adipose conversion. A similar dependence was observed with drug-treated Ob17MT18 and 3T3-F442A preadipocyte cells, but not with non-differentiating 3T3-C2 cells. Studies in drug-treated Ob1771 cells at the mRNA level showed that the parallel expression of mRNAs encoding for glycerol-3-phosphate dehydrogenase and an homologue of serine proteinases of Mr 28,000 [Cook, Groves, Min & Spiegelman (1985) Proc. Natl. Acad. Sci. U.S.A. 82, 6480-6484] was also dependent on putrescine addition. Double-isotope experiments with [14C]putrescine and [3H]spermidine, as well as analysis of the polyamine content in drug-treated Ob1771 cells under various conditions, demonstrate after putrescine addition that the expression of late markers of adipose conversion was highly correlated with a 2-fold increase in the intracellular concentration of spermidine. No correlation was observed with changes in the intracellular concentrations of putrescine and spermine. Long-term exposure of untreated Ob1771 cells to growth hormone, which led to the expression of late markers of adipose conversion [Doglio, Dani, Grimaldi & Ailhaud (1986) Biochem. J. 238, 123-129] was also accompanied by the same increase in spermidine concentration, which attained values identical with those determined in drug-treated cells supplemented with putrescine. This observation suggests that the permissive effect of growth hormone on the terminal differentiation of adipose cells might e related to changes in the intracellular concentration of spermidine.

Activity of human growth hormone and related polypeptides on the adipose conversion of 3T3 cells

1984 ◽  
Vol 4 (2) ◽  
pp. 228-231
Author(s):  
M Morikawa ◽  
H Green ◽  
U J Lewis
Keyword(s):  
Growth Hormone ◽  
Human Growth Hormone ◽  
Culture System ◽  
Human Growth ◽  
3T3 Cells ◽  
Specific Assay ◽  
Established Line ◽  
Adipose Conversion ◽  
Cellular Responsiveness ◽  
Adipose Cells

A culture system is described for the study of cellular responsiveness to growth hormone. The hormone acts directly on an established line of preadipose 3T3 cells and promotes their differentiation into adipose cells. This response is the basis of a sensitive and specific assay and does not depend on the participation of an intermediate effector.

scholarly journals Growth hormone regulation of the expression of differentiation-dependent genes in preadipocyte Ob1771 cells

1986 ◽  
Vol 238 (1) ◽  
pp. 123-129 ◽  
Author(s):  
A Doglio ◽  
C Dani ◽  
P Grimaldi ◽  
G Ailhaud
Keyword(s):  
Growth Hormone ◽  
Mrna Level ◽  
Protein A ◽  
Transcriptional Level ◽  
Hormone Regulation ◽  
Beta Actin ◽  
Adipose Conversion ◽  
Specific Mrnas ◽  
Actin Mrna ◽  
Glycerol 3 Phosphate Dehydrogenase

The adipose conversion of Ob1771 preadipocytes, during exposure to medium containing bovine serum and supplemented with growth hormone, is accompanied by the acquisition of phenotypic markers and the increased accumulation of specific mRNAs. The expression of lipoprotein lipase, and that of unidentified pOb24 and pGH3 mRNAs, are early events which are independent of growth hormone supplementation. By contrast, the late expression of mRNAs encoding for glycerol-3-phosphate dehydrogenase and p422 protein (a myelin-P2 homologue) and that of glycerol-3-phosphate dehydrogenase activity require the presence of growth hormone. The abundance of beta-actin mRNA does not change during differentiation. Runoff transcription by nuclei isolated from untreated or growth hormone-treated cells reveal little or no change in the rates of transcription of pOb24, pGH3 and beta-actin mRNAs. By contrast, the transcription rate of the p422 gene increases markedly (greater than 6-fold) in nuclei of growth hormone-treated cells. However, the p422 mRNA is more abundant than would be predicted by its nuclear transcription alone, suggesting, in Ob1771 cells exposed to growth hormone, that there is a post-transcriptional level of control. These results indicate that the permissive role of growth hormone during adipose cell differentiation is related to terminal events only and that its effects can be seen both at the protein and mRNA level. These results strongly suggest that an increased rate of specific transcription is primarily responsible for the accumulation of mRNAs during exposure to growth hormone.

scholarly journals Activity of human growth hormone and related polypeptides on the adipose conversion of 3T3 cells.

1984 ◽  
Vol 4 (2) ◽  
pp. 228-231 ◽  
Author(s):  
M Morikawa ◽  
H Green ◽  
U J Lewis
Keyword(s):  
Growth Hormone ◽  
Human Growth Hormone ◽  
Culture System ◽  
Human Growth ◽  
3T3 Cells ◽  
Specific Assay ◽  
Established Line ◽  
Adipose Conversion ◽  
Cellular Responsiveness ◽  
Adipose Cells

A culture system is described for the study of cellular responsiveness to growth hormone. The hormone acts directly on an established line of preadipose 3T3 cells and promotes their differentiation into adipose cells. This response is the basis of a sensitive and specific assay and does not depend on the participation of an intermediate effector.

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