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Biomedical subjects

G Ailhaud

Publications and source records attributed to G Ailhaud.

At least 73 records · Page 4Linked to original sources
PubMed1993

Inhibition of prostacyclin-induced Ca2+ mobilization by phorbol esters in Ob1771 preadipocytes.

In addition to cAMP production, a transient elevation of intracellular free Ca2+ has been shown to take place in preadipose cells upon stimulation by carbaprostacyclin (cPGI2), both messengers acting in synergy to initiate adipose cell differentiation (Vassaux, G., Gaillard, D., Ailhaud, G., and Négrel, R. (1992) J. Biol. Chem.267, 11092-11097). Further studies reported herein show that this Ca2+ transient is i) elicited by the natural prostaglandin PGI2, ii) independent of the presence of extracellular Ca2+, suggesting a mobilization of Ca2+ from intracellular pools and ii) unaffected by cAMP elevating agents. Moreover, and in contrast to the InsP3-dependent Ca2+ signal evoked by PGF2 alpha, that induced by PGI2 is fully abolished by pretreatment with phorbol esters (EC50: 1-5 nM). Furthermore, experiments designed to empty the Ca2+ pools, using PGI2 or PGF2 alpha as Ca2+ mobilizing agents as well as pretreatments with drugs, allow to conclude that PGI2 mobilizes Ca2+ from an InsP3 sensitive, ryanodine insensitive intracellular pool. Altogether, these results strongly suggest that PGI2 mobilizes Ca2+ from an intracellular store common to that affected by InsP3, by means of a mechanism which remains to be elucidated.

1-Methyl-3-isobutylxanthine↗
PubMed1993

Terminal differentiation of mouse preadipocyte cells: adipogenic and antimitogenic role of triiodothyronine.

The role of triiodothyronine (T3) in the differentiation process of Ob1771 mouse preadipocyte cells has been studied under serum-free and hormone supplemented culture conditions which were previously shown to lead to terminal differentiation. In the absence of T3, a dramatic decrease in the adipogenic activity of the culture medium (EC50 = 0.1 nM) could be observed, as indicated 12 days after confluence by the low levels of late markers of differentiation such as adipsin, lipid-binding protein aP2 and glycerol-3-phosphate dehydrogenase as well as the sharp reduction of the number of triacyglycerol-containing cells. This decrease in adipogenic activity was accompanied by a parallel increase of the mitogenic potency of the culture medium. Therefore, T3 appears to be a hormone capable of modulating both proliferation and differentiation of preadipocytes. T3 ceased to be necessary provided the culture medium was supplemented with high concentrations of inducers of differentiation, such as 8-bromo-cAMP or carbaprostacyclin.

1-Methyl-3-isobutylxanthine↗
PubMed1993

Characterization of apoA-IV-containing lipoprotein particles isolated from human plasma and interstitial fluid.

Apolipoprotein (apo) A-IV has been proposed to play a role in reverse cholesterol transport. ApoA-IV-containing lipoprotein particles (A-IVLp) were isolated from human plasma and interstitial fluid and characterized by immunoaffinity chromatography. Two major A-IVLp subpopulations, lipoprotein particles containing apoA-IV with apoA-I (LpA-I:A-IV) and lipoprotein particles containing apoA-IV without apoA-I (LpA-IV), were identified. The larger subpopulation of A-IVLp is the LpA-IV that represents 70% (protein mass) of the initial particles. Only 5.8% of apoA-IV was recovered in the retained fraction after affinity chromatography with an anti-apoA-I immunosorbent. ApoA-I, apoA-II, apoA-IV, apoB, apoC-III, apoD, apoE, apoH, lecithin: cholesterol acyltransferase (LCAT), cholesteryl ester transfer (CET) protein, proline-rich protein, and a protein of Mr 59,000 were detected in the A-IVLp. These particles contain more than 20% triglycerides (lipid mass). ApoA-IV-containing particles that were isolated from plasma are heterogeneous in size, consisting of two major populations with Stokes' diameters of 10.3 nm and 9.3 nm. Both subpopulations of A-IVLp contain LCAT and CET activities and promote cholesterol efflux from cholesterol-preloaded adipose cells. These data support the hypothesis that A-IVLp particles may be involved in reverse cholesterol transport.

Adipose Tissue↗
PubMed1993

The regulation by growth hormone of lipoprotein lipase gene expression is mediated by c-fos protooncogene.

GH has been previously shown in Ob1771 adipose cells to activate transiently the expression of c-fos gene by a protein kinase-C-dependent pathway and to modulate, at last in part by a protein kinase-C-dependent pathway, the expression of the lipoprotein lipase (LPL) gene. In Ob1771 cells exposed to GH, under conditions where protein synthesis is inhibited by cycloheximide, the modulation of LPL gene expression is prevented, suggesting that synthesis of trans-acting factor(s) is required to modulate LPL gene expression. The present results indicate the involvement of c-Fos protein in this modulation; this involvement is supported by various lines of evidence: 1) upon GH stimulation, the increase in c-fos mRNA content is followed by the emergence of c-Fos protein within the nucleus, and this emergence precedes the increase in LPL mRNA content; 2) in GH-treated Ob1771 cells, exposure to antisense sof oligonucleotides abolishes the synthesis of c-Fos protein; and 3) at the same time, the increase in LPL mRNA content and LPL activity does not occur, whereas sense fos oligonucleotides show no effect. It is concluded that c-Fos protein plays an intermediary role in the modulation of LPL gene expression by GH.

Adipose Tissue↗
PubMed1993

Heterogeneity of high density lipoprotein particles.

Human high density lipoprotein (HDL) contains at least two types of lipoprotein-containing A-I particles: one contains both apolipoproteins (apo) A-I and A-II (Lp A-I:A-II) as main protein components, whereas in the other, apo A-II is absent (Lp A-I). The two subpopulations have different metabolisms and physiological roles. Studies indicate that apo A-II is an antagonist of cholesterol efflux. Lp A-I but not Lp A-I:A-II is increased in women compared with men. The lower apo A-I levels in patients with coronary artery disease reflect a decrease in Lp A-I. Lp A-I also is decreased in children with a family history of coronary artery disease, and increased in elderly subjects. Despite cholesterol intake remaining constant, a high polyunsaturated:saturated fat diet leads to a decrease in Lp A-I but not Lp A-I:A-II. Chronic alcohol consumption induces an increase in Lp A-I:A-II and a decrease in Lp A-I, although short bouts of drinking produce increases in both Lp A-I and Lp A-I:A-II. Hypolipidemic drugs have specific effects on discrete apo A-I particles.

Alcoholism↗
PubMed1992

Essential role of collagens for terminal differentiation of preadipocytes.

In order to study the role of collagens in the differentiation of TA1 preadipose cells in vitro, ethyl-3,4-dihydroxybenzoate (EDHB) was used as a specific inhibitor of collagen synthesis. The secretion of collagenous proteins only was severely decreased after exposure to EDHB, and this was accompanied by a decrease of differentiation as indicated by low activity levels of glycerophosphate dehydrogenase. The effect of EDHB was dose-dependent and also dependent upon the stage of cell differentiation. Northern-blot analysis show that EDHB addition to undifferentiated cells did not prevent the induction of A2COL6 gene, a marker of the preadipose state, but prevented the induction of the gene encoding for the adipocyte lipid binding protein and the modulation of the expression of the lipoprotein lipase gene which are both indicators of the adipose state. These results demonstrate that differentiation of preadipose cells into adipose cells requires active synthesis of collagens during the preadipose state.

Adipose Tissue↗
PubMed1992

Prostacyclin is a specific effector of adipose cell differentiation. Its dual role as a cAMP- and Ca(2+)-elevating agent.

The mitogenic-adipogenic activity of carbaprostacyclin (cPGI2), a stable analogue of prostacyclin (PGI2), has been proposed to be related to its ability to elicit cAMP production and to activate the protein kinase A cascade (Négrel, R., Gaillard, D., and Ailhaud, G. (1989) Biochem. J. 257, 399-405). In the present study, cPGI2 has been compared with other activators of the cAMP pathway, namely isoproterenol, forskolin and 8-bromo-cAMP, with respect to adipose cell differentiation. Carbaprostacyclin behaved as a much more potent and efficient effector of mouse Ob1771 preadipocyte differentiation than the latter agents. Moreover, cPGI2 also exerted a specific amplifying mitogenic-adipogenic role, as compared with isoproterenol in rat as well as human adipose precursor cells in primary culture, suggesting that the prostanoid was able to generate an additional second messenger. The fact that ionomycin was able to potentiate and amplify the differentiation induced by 8-bromo-cAMP led us to give evidence, using preadipocytes preloaded with the fluorescent calcium chelator Indo-1, that cPGI2, besides its ability to activate adenyl cyclase, was also able to induce a transient increase in intracellular free calcium. This phenomenon was independent of cAMP production or inositol phospholipid breakdown and appeared to be mediated after binding to a single class of PGI2 receptor. The potential to generate simultaneously two synergistic intracellular signals allows us to ascribe to PGI2 a key and specific role in the differentiation of adipose precursor cells in vitro that would likely lead in vivo to the recruitment of "dormant" preadipocytes to become adipocytes.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1992

Cellular and molecular aspects of adipose tissue development.

Both in animals and humans, before or after birth, angiogenesis appears to be closely coordinated in time and space with the formation of fat cell clusters. Monobutyrin, a novel fat-specific angiogenesis factor, may play a role in this process. The potential to acquire new fat cells appears to be permanent throughout life in both animals and humans, as revealed by in vitro experiments. Considerable evidence now supports the view that BAT and WAT are distinct organs; in addition, the existence of distinct BAT precursor cells is demonstrated by their unique ability to express the UCP gene. In bovine and ovine, the transformation of BAT into WAT is strongly suggested by the rapid disappearance after birth of UCP from the various BAT depots. Despite the initial cell heterogeneity of the stromal-vascular fraction, cultured stromal-vascular cells of adipose tissue are adipose precursor cells that show varying capacities for replication and differentiation, according to age and fat depot. Studies of adipose cell differentiation in vitro correspond to the sequence: adipoblast (unipotential cells)----commitment preadipose cell (preadipocyte)----terminal differentiation immature adipose cell----terminal differentiation mature adipose cell (adipocyte). Cell commitment is triggered by growth arrest and characterized by the expression of early markers (A2COL6/pOb24; clone 5; LPL), whereas only terminal differentiation of preadipocytes requires the presence of various hormones. Multiple signaling pathways have been characterized and shown to cooperate in the process of terminal differentiation. The concept that adipose cells behave as secretory cells is now emerging from in vitro data, since secretion of various proteins (LPL, adipsin, CETP) and important metabolites (fatty acids, monobutyrin, androgens, estrogens, prostaglandins) takes place both constitutively and upon hormonal stimulation. This suggests that adipose tissue participates more directly than previously thought in metabolic activities and energy balance.

Adipose Tissue↗
PubMed1992

Differential response of preadipocytes and adipocytes to prostacyclin and prostaglandin E2: physiological implications.

The major prostaglandins (PGs) locally produced in adipose tissue both in rodent and man are PGE2 and prostacyclin (PGI2). We have recently described PGI2 as an autocrine promoter and/or amplifier of terminal differentiation of cultured preadipocytes in several species. The effectiveness and specificity of PGI2 as an adipogenic agent are related to its ability to induce in preadipocytes intracellular increases of both cAMP and free calcium. Moreover, PGs of the E series are well known to exert an antilipolytic effect in mature adipocytes. These observations have prompted us to address two questions of physiological interest: 1) Is PGI2 still able to increase cAMP in differentiated adipocytes, behaving thus as a lipolytic agent, and 2) Is PGE2 able to negatively modulate cAMP production in adipose precursor cells, behaving thus as a counteracting effector of PGI2 action? Our results, with respect to cAMP production and/or lipolysis and antilipolysis, demonstrate clearly that in adipose tissue of both rat and man, PGI2 exclusively affects adipose precursor cells whereas PGE2 exclusively affects adipocytes. We propose a model of concerted action for both PGs in the development of adipose tissue mass, PGI2 behaving as an adipogenic-hyperplastic effector and PGE2 as an antilipolytic-hypertrophic effector.

Adipose Tissue↗
PubMed1992

Apolipoprotein A-I-containing particles and reverse cholesterol transport in IDDM.

Lipoprotein particles containing apoA-I but not apoA-II are, among high-density lipoproteins, effective protectors against atherosclerosis that act by promoting the efflux of cellular cholesterol and the reverse cholesterol transport process. Because previous studies showed that in vitro nonenzymatic glycosylation of HDL impairs HDL receptor-mediated cholesterol efflux, we isolated Lp A-I from two poorly controlled insulin-dependent diabetic patients and compared the chemical composition and ability to promote cholesterol efflux with the same particles purified from two matched nondiabetic control subjects. No differences in lipid composition or in the ability to promote cholesterol efflux from cultured adipose cells were noted between the two types of Lp A-I preparations. However, when we separated Lp A-I from diabetic subjects by degree of glycosylation, the specifically glycosylated subfractions were about 50% less effective in producing cholesterol efflux than the nonglycosylated particles.

Adult↗
PubMed1992

Terminal differentiation of mouse preadipocyte cells: the mitogenic-adipogenic role of growth hormone is mediated by the protein kinase C signalling pathway.

The role of growth hormone (GH) in the differentiation process of Ob1771 mouse preadipocyte cells has been studied under culture conditions that were serum-free and hormone-supplemented and which were previously shown to lead to terminal differentiation. In the absence of GH, a dramatic decrease in the adipogenic activity of the culture medium could be observed, as indicated 12 days after confluence by the low levels of glycerol-3-phosphate dehydrogenase activity and the sharp reduction of the number of triacylglycerol-containing cells. This decrease in adipogenic activity was accompanied by a parallel loss of the mitogenic potency of the culture medium. Determination of the half-maximal and maximal concentrations of GH required for the restoration of growth and differentiation were identical, 0.5 and 2 nM, respectively. Despite the presence of insulin-like growth factor-I (IGF-I) to substitute for supraphysiological concentrations of insulin and to saturate IGF-I receptor, GH was still required to induce terminal differentiation of a maximal number of cells. However, protein kinase C activators such as prostaglandin F2 alpha, phorbol esters and diacylglycerol were able to mimic GH in promoting a maximal mitogenic-adipogenic response, indicating that the ability of GH to induce diacylglycerol production (Doglio et al., 1989; Catalioto et al., 1990) plays a prominent role in this process. Furthermore, in agreement with the fact that the mitoses which precede terminal differentiation of Ob1771 preadipocytes are strictly controlled by cAMP and only modulated by protein kinase C, terminal differentiation of Ob1771 preadipocytes occurred in the absence of GH upon supplementation with high concentrations of carbaprostacyclin, added as a cAMP-elevating agent or with 8-Br-cAMP, added as a cAMP analogue. It is concluded that the control exerted by GH on terminal differentiation of mouse preadipocytes corresponds to a modulating mitogenic effect mediated through protein kinase C activation and leading to a potentiation of the cAMP and IGF-I mitogenic signalling pathways.

Adipose Tissue↗
PubMed1992

Apolipoprotein A-containing lipoprotein particles: physiological role, quantification, and clinical significance.

High-density lipoprotein (HDL) particles are made up of two major populations of particles, differing in composition and metabolism. Both contain apolipoprotein (apo) A-I but only one contains apo A-II. Lipoprotein particles that contain only apo A-I (LpA-I particles) can increase cellular cholesterol efflux from cultured cells in vitro. LpA-I:A-II particles, however, do not increase cholesterol efflux. LpA-I:A-II can be determined directly with an enzyme-linked differential antibody immunosorbent assay. LpA-I is determined by differential electroimmunoassay: in the presence of a large excess of anti-apo A-II, LpA-I:A-II particles are retained in one peak, whereas LpA-I migrates grates as a second peak. Both lipoprotein forms of apo A-I-containing particles are present mainly in HDL, but the relative proportion of LpA-I is greater in HDL2 than in HDL3. Concentrations of LpA-I in plasma samples from normolipemic subjects average approximately 10% higher in women than in men. The lower apo A-I concentrations in patients with significant coronary artery disease reflect a decrease in the LpA-I particles. Data obtained in octogenarians also support the possibility that LpA-I might represent the anti-atherogenic fraction of HDL. Moreover, the concentration of LpA-I in children of parents with premature coronary heart disease was lower than that of a control group without any family history of this disease. Nutrients and hypolipidemic drugs seem to affect the two kinds of particles differently.

Adipose Tissue↗
PubMed1991

Autocrine control of adipose cell differentiation by prostacyclin and PGF2 alpha.

The mitogenic-adipogenic effect exerted by arachidonic acid, which leads to terminal differentiation of Ob1771 mouse preadipocytes, has been shown to be (i) blocked by cyclooxygenase inhibitors, (ii) mimicked by a stable analogue of prostacyclin (carbaprostacyclin) and (iii) potentiated by PGF2 alpha. Since these prostanoids are known to be synthesized and secreted by preadipocytes, we have proposed that both prostacyclin as the key mediator and PGF2 alpha as a modulator control the expression of terminal events of adipose conversion by means of an autocrine mechanism (Gaillard, D. et al. and Negrel, R. et al. Biochem. J. (1989) 257, 389-397 and 399-405). In order to test this hypothesis, the release of prostacyclin, characterized under the form of its stable degradation product 6-keto-PGF1 alpha, and that of PGF2 alpha have been studied in the culture medium of Ob1771 cells. A striking increase in the release of 6-keto-PGF1 alpha and to a minor degree of PGF2 alpha was observed when cells were exposed to arachidonic acid as shown by using [3H]arachidonic acid prelabelled cells or by radio-immunoassays. Since antagonists of PGF2 alpha and PGI2 receptors were not available, specific antibodies directed against PGF2 alpha and 6 beta-PGI1, another stable analogue of prostacyclin, were added as neutralizing agents in the culture medium. These antibodies were able to counteract the mitogenic-adipogenic effect of arachidonic acid. Prostacyclin and PGF2 alpha thus appear as autocrine mediators in the process of adipose conversion.

Adipose Tissue↗