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S Spiegel

Publications and source records attributed to S Spiegel.

At least 109 records · Page 6Linked to original sources
PubMed1996

A new wound healing agent--sphingosylphosphorylcholine.

Spingosylphosphorylcholine (lysosphingomyelin or SPC) is an effective and broad spectrum cell growth promoting agent and a candidate for evaluation on wound healing. The effect of SPC on full-thickness excision and incision wounds in genetically healing-impaired diabetic (db/db) mice was evaluated by measurement of wound area, skin strength, and tissue histology. The effect on cell proliferation was measured in vivo by incorporation of bromo-deoxyuridine and in vitro by [3H] thymidine incorporation. SPC increased the rate of wound closure, with a statistically significant improvement in measured wound areas (p < 0.02, compared with vehicle controls). The optimum concentration was 2-3 microM. SPC, alone and in combination with insulin, stimulated DNA synthesis in cells known to participate in wound healing, including microvascular endothelial cells. In vivo, SPC stimulated proliferation of keratinocytes, fibroblasts, endothelial cells, and cells around sebaceous glands and hair follicles at day 2-4 postwound, resulting in a complete re- epithelialization and profound granulation tissue formation in excisional and incisional wound sites of db.db and db/+ mice. Quantitative assessment of wound tissue section morphology indicated that SPC induced up to a 3-fold increase in the numbers of mitotic cells, resulted in smaller cross-sectional scar area, and led to more normalized tissue in the wound sites. SPC had no deleterious effect on wound skin strength. In conclusion, the acceleration of dermal wound healing animal models suggests that SPC could be an interesting candidate for clinical application.

Animals↗
PubMed1996

Structural requirements of sphingosylphosphocholine and sphingosine-1-phosphate for stimulation of activator protein-1 activity.

The sphingolipids sphingosine-1-phosphate (SPP) and sphingosylphosphocholine (SPC) stimulate mitogenesis in Swiss 3T3 fibroblasts and stimulate DNA binding activity of activator protein-1 (AP-1). We show that SPP and SPC were more potent agents than nonphosphorylated sphingosines and N-acyl-sphingolipids (ceramides) with respect to DNA synthesis, AP-1 DNA binding activity, and AP-1 trans-activation, illustrating the importance of the terminal phosphate group. The free 2-amino group and the 4E double bond of SPC and SPP were found to be important for these activities. Although the combination of decreasing the sphingoid backbone chain length of SPC by two carbons and hydrogenating the 4E bond only slightly reduced its effects, in contrast, the same modifications in SPP significantly decreased its mitogenic and AP-1 trans-activation effects. Furthermore, substitution of the 3-hydroxyl group in SPP with hydrogen decreased its ability to stimulate DNA synthesis and to stimulate AP-1 transcriptional activity. Thus, critical sphingolipid structural components for AP-1 activation and mitogenic stimulation include the free 2-amino group, the free 3-hydroxyl group, the 4,5-trans double bond, and terminal phosphorylation. These observations may be relevant for clinical uses of these compounds in applications such as wound healing and inhibition of metastasis.

3T3 Cells↗
PubMed1995

Sphingosylphosphocholine, a signaling molecule which accumulates in Niemann-Pick disease type A, stimulates DNA-binding activity of the transcription activator protein AP-1.

Sphingosylphosphocholine (SPC) is the deacylated derivative of sphingomyelin known to accumulate in neuropathic Niemann-Pick disease type A. SPC is a potent mitogen that increases intracellular free Ca2+ and free arachidonate through pathways that are only partly protein kinase C-dependent. Here we show that SPC increased specific DNA-binding activity of transcription activator AP-1 in electrophoretic mobility-shift assays. Increased DNA-binding activity of AP-1 was detected after only 1-3 min, was maximal after 6 hr, and remained elevated at 12-24 hr. c-Fos was found to be a component of the AP-1 complex. Northern hybridization revealed an increase in c-fos transcripts after 30 min. Since the increase in AP-1 binding activity preceded the increase in c-fos mRNA, posttranslational modifications may be important in mediating the early SPC-induced increases in AP-1 DNA-binding activity. Western analysis detected increases in nuclear c-Jun and c-Fos proteins following SPC treatment. SPC also transactivated a reporter gene construct through the AP-1 recognition site, indicating that SPC can regulate the expression of target genes. Thus, SPC-induced cell proliferation may result from activation of AP-1, linking signal transduction by SPC to gene expression. Since the expression of many proteins with diverse functions is known to be regulated by AP-1, SPC-induced activation of AP-1 may contribute to the pathophysiology of Niemann-Pick disease.

3T3 Cells↗
PubMed1995

The role of calcium influx in cellular proliferation induced by interaction of endogenous ganglioside GM1 with the B subunit of cholera toxin.

The B subunit of cholera toxin, which binds specifically to ganglioside GM1, is mitogenic for quiescent Swiss 3T3 fibroblasts. It was previously shown that the B subunit had no effect on cAMP, protein kinase C or phosphoinositide turnover, but did cause an increase in the influx of calcium from extracellular sources (Spiegel, S. and Panagiotopoulos, C. (1988) Exp. Cell Res. 177, 414-427). In contrast to the action of known growth factors, the B subunit induced significant DNA synthesis after only a 1-3 h treatment. We utilized this unique property to determine whether the increase in calcium influx plays a role in B subunit-induced mitogenicity. Cells were briefly treated with the B subunit in the presence of calcium channel blockers, followed by removal of the blockers and further incubation in B subunit-free medium for the remaining time required to measure DNA synthesis. When 1 mM cobalt was only present during the first 3 h incubation. DNA synthesis induced by either the B subunit or fetal bovine serum was completely abolished. However, both nickel (1 mM) adn the L-type voltage-gated calcium channel inhibitor nicardipin (10 microM) inhibited B subunit-induced cell proliferation without abrogating the response to fetal bovine serum. Using a gel retardation assay, we found that the B subunit markedly stimulated specific DNA-binding activity of the transcription factor, activator protein-1 (AP-1), which functions as a major convergence point coupling early events induced by a variety of mitogens to long term growth responses. Presence of c-Fos protein in the AP-1 complex was demonstrated as a supershift band in the gel mobility assay using c-Fos polyclonal antibody. Cobalt, which markedly inhibited B subunit-induced DNA synthesis, also completely abolished AP-1 DNA-binding activity stimulated by the B subunit. In sharp contrast, cobalt had no effect on DNA-binding activity of AP-1 induced by the tumor promoter, 12-O-tetradecanoylphorbol 13-acetate. Our results suggest that calcium influx is a key element for both DNA-binding activity of AP-1 and cell proliferation induced by binding of the B subunit of cholera toxin to cell surface ganglioside GM1.

3T3 Cells↗
PubMed1995

Sphingosine 1-phosphate rapidly activates the mitogen-activated protein kinase pathway by a G protein-dependent mechanism.

Addition of sphingosine 1-phosphate induces proliferation of quiescent Swiss 3T3 fibroblasts by unknown mechanisms. To identify the pathways involved, the ability of sphingosine 1-phosphate to activate mitogen-activated protein (MAP) kinase was studied. Sphingosine 1-phosphate rapidly activated the Raf/MAP kinase kinase (MKK)/MAP kinase pathway, and the concentration dependence for MAP kinase activation correlated with that for induction of DNA synthesis. Both MKK1 and MKK2 were activated by sphingosine 1-phosphate, assessed by specific immune complex kinase assays. Prior treatment of the Swiss 3T3 cells with pertussis toxin inhibited 70-80% of the sphingosine 1-phosphate-stimulated MAP kinase activity. Thus, one of the direct or indirect targets of exogenous sphingosine 1-phosphate appears to be a G(i)/G(o) protein.

3T3 Cells↗
PubMed1995

Involvement of a pertussis toxin-sensitive G protein in the mitogenic signaling pathways of sphingosine 1-phosphate.

Sphingosine 1-phosphate, a sphingolipid metabolite, was previously reported to increase DNA synthesis in quiescent Swiss 3T3 fibroblasts and to induce transient increases in intracellular free calcium (Zhang, H., Desai, N. N., Olivera, A., Seki, T., Brooker, G., and Spiegel, S. (1991) J. Cell Biol. 114, 155-167). In the present study, pretreatment of Swiss 3T3 fibroblasts with pertussis toxin reduced sphingosine 1-phosphate-induced DNA synthesis. Sphingosine 1-phosphate decreased cellular cAMP levels and also caused a drastic decrease in isoproterenol- and forskolin-stimulated cAMP accumulation. Pertussis toxin treatment prevented the inhibitory effect of sphingosine 1-phosphate on cAMP accumulation, suggesting that a pertussis toxin-sensitive Gi or Gi-like protein may be involved in sphingosine 1-phosphate-mediated inhibition of cAMP accumulation. Mitogenic concentrations of sphingosine 1-phosphate stimulated production of inositol phosphates which was inhibited by pertussis toxin, while the response to bradykinin was not affected. Furthermore, calcium release induced by sphingosine 1-phosphate, but not by bradykinin, was also attenuated by pertussis toxin treatment. However, sphingosine 1-phosphate-induced phosphatidic acid accumulation was unaffected by pertussis toxin. The increase in specific DNA binding activity of activator protein-1, which was induced by treatment of quiescent Swiss 3T3 fibroblasts with sphingosine 1-phosphate, was also inhibited by pertussis toxin. These results suggest that some of the sphingosine 1-phosphate-induced signaling pathways are mediated by G proteins that are substrates for pertussis toxin.

3T3 Cells↗
PubMed1994

Stereospecificity of sphingosine-induced intracellular calcium mobilization and cellular proliferation.

Sphingosine is a positive regulator of cell growth in Swiss 3T3 fibroblasts (Zhang, H., Buckley, N. E., Gibson, K., and Spiegel, S. (1990) J. Biol. Chem. 265, 76-81). The present study investigated the stereospecificity of sphingosine-induced cell proliferation and its mitogenic signal transduction mechanisms. D-(+)-erythro Stereoisomers (cis and trans) stimulated DNA synthesis, whereas neither L-(-)-threo-sphingosine (cis or trans) nor DL-threo-dihydrosphingosine had any effect. Previously, we have shown that sphingosine-1-phosphate may mediate the mitogenic effect of sphingosine (Zhang, H., Desai, N. N., Olivera, A., Seki, T., Brooker, G., and Spiegel, S. (1991) J. Cell Biol. 114, 155-167). However, no major differences were found in the formation of D-(+)-erythro- and L-(-)-threo- sphingosine-1-phosphate derived from the respective sphingosine isomers in intact cells. Thus, the stereospecificity of the response to sphingosine may reside at the level of specific intracellular targets for sphingosine-1-phosphate. Sphingosine-1-phosphate triggers dual signal transduction pathways of activation of phospholipase D leading to increases in the levels of phosphatidic acid and mobilization of calcium from internal stores. Both D-(+)-erythro- and L-(-)-threo-sphingosine isomers induced similar increases in phosphatidic acid concomitant with identical decreases in phosphatidylcholine levels. In contrast, only the D-(+)-erythro-stereoisomers (cis and trans) were effective in releasing calcium from intracellular stores. Our results suggest that the formation of phosphatidic acid is not sufficient to mediate sphingosine-stimulated DNA synthesis. However, the stereospecificity of the sphingosine-induced mobilization of calcium from internal stores seems to correlate with the induction of DNA synthesis by sphingosine stereoisomers.

3T3 Cells↗
PubMed1994

Sphingosine 1-phosphate, a novel signaling molecule, stimulates DNA binding activity of AP-1 in quiescent Swiss 3T3 fibroblasts.

Sphingosine and sphingosine 1-phosphate, metabolites of sphingolipids, stimulate cell proliferation in quiescent Swiss 3T3 fibroblasts and induce transient increases in intracellular free calcium (Zhang, H., Desai, N. N., Olivera, A., Seki, T., Brooker, G., and Spiegel, S. (1991) J. Cell Biol. 114, 155-167). However, little is yet known of the nuclear events that follow the early responses induced by sphingolipid metabolites. Using a gel retardation assay, we found that specific DNA binding activity of activator protein-1 (AP-1) was markedly increased after treatment of quiescent Swiss 3T3 fibroblasts with sphingosine 1-phosphate and sphingosine. The DNA binding specificity of AP-1 was confirmed with competing probes containing consensus sequences of AP-1, AP-2, AP-3, SP-1, and NF1/CTF. The c-fos gene product was detected in the AP-1 complex using anti-c-Fos antibody. The dose response for stimulation of DNA binding activity of AP-1 by sphingosine 1-phosphate correlated closely with its effect on DNA synthesis. Furthermore, an inhibitor of sphingosine kinase, DL-threo-dihydrosphingosine, which inhibits sphingosine-induced DNA synthesis and the formation of sphingosine 1-phosphate, also inhibited sphingosine-stimulated AP-1 DNA binding activity. This result further supports our proposal that sphingosine 1-phosphate mediates the mitogenic effect of sphingosine. Our results indicate that sphingosine 1-phosphate-induced DNA synthesis and cell division may result from activation of AP-1 protein, linking signal transduction by sphingolipid metabolites to gene expression.

3T3 Cells↗
PubMed1994

Glycosphingolipids of rat T cells. Predominance of asialo-GM1 and GD1c.

Glycosphingolipids play an important role in the immune response, yet their compositions in T and B cells which mediate cellular and humoral immunity, respectively, have not been elucidated. In this study, characteristic features of glycosphingolipids in rat T lineage cells were revealed by comparing the gangliosides and neutral glycolipids of spleen T- and beta-cell-enriched fractions and thymocytes. In T cells, GD1c(NeuGc,NeuGc), a unique ganglioside synthesized through asialo-GM1 (GA1), was the predominant ganglioside as previously found in thymocytes [Nohara, K., Suzuki, M., Inagaki, F., & Kaya, K. (1991) J. Biochem. (Tokyo) 110, 274-278], and the amount was much higher than in thymocytes. In addition, three other GA1-derived gangliosides were detected in T cells and identified as GM1b(NeuAc), GM1b(NeuGc), and GD1 alpha(NeuAc,NeuAc). In contrast, GD1 alpha(NeuAc,NeuAc) was not discernible in thymocytes, although gangliosides corresponding to GM1b(NeuAc) and GM1b(NeuGc) were detected. The neutral glycolipids of T cells contained almost exclusively GA1, while thymocytes contained much lower amounts. The predominance of these GA1-derived gangliosides was confirmed as a singular feature of T lineage cells by comparison with gangliosides of spleen B-cell-enriched fractions which mainly consisted of gangliosides synthesized through GM3 and GM1. Furthermore, the unique structures, which contain the GM1 core and the extended modification of the lacto series, alpha Gal-LacNAc-GM1, alpha Gal-(LacNAc)2-GM1, and sialyl-LacNAc-GM1, were found in B-cell-enriched fractions. Unexpectedly, the neutral glycolipid composition of the thymocytes resembled that of the B-cell enriched fraction rather than that of the T cells.

Animals↗
PubMed1994

Sphingosine-1-phosphate, a putative second messenger, mobilizes calcium from internal stores via an inositol trisphosphate-independent pathway.

Sphingosine-1-phosphate, a metabolite of sphingolipids which has previously been shown to stimulate DNA synthesis and cell division in quiescent cultures of Swiss 3T3 fibroblasts (Zhang, H., Desai, N. N., Olivera, A., Seki, T., Brooker, G., and Spiegel, S. (1991) J. Cell Biol. 114, 155-167), induced a transient increase in intracellular free calcium independent of extracellular calcium. The increase in calcium was completely abolished when intracellular calcium pools were depleted with thapsigargin, an inhibitor of the endoplasmic reticulum Ca(2+)-ATPase. The dose-response for calcium release induced by sphingosine-1-phosphate correlated closely with the concentration required for stimulation of DNA synthesis. The magnitude of the calcium response decreased with successive challenges, although sphingosine-1-phosphate did not attenuate the responses to either bradykinin or ionomycin. Conversely, prior stimulation of the cells with bradykinin had no effect on the sphingosine-1-phosphate-induced calcium signal. Although sphingosine-1-phosphate increased inositol (1,4,5)-trisphosphate levels, complete inhibition of inositol phosphate formation by pretreatment with 12-O-tetradecanoylphorbol-13-acetate did not block sphingosine-1-phosphate-mediated calcium responses. Moreover, in permeabilized cells, heparin, an inositol (1,4,5)-trisphosphate antagonist, blocked Ca2+ release induced by inositol (1,4,5)-trisphosphate, but did not significantly alter the Ca2+ release induced by sphingosine-1-phosphate. Sphingosine-1-phosphate did not stimulate the release of arachidonic acid, another signaling molecule known to elevate [Ca2+]i without inositol lipid turnover or calcium influx. Our data suggest that sphingosine-1-phosphate mobilizes Ca2+ from internal stores primarily through a mechanism independent of inositol lipid hydrolysis and arachidonic acid release and that sphingolipid metabolism may be important in calcium homeostasis.

3T3 Cells↗
PubMed1994

Sphingosine kinase from Swiss 3T3 fibroblasts: a convenient assay for the measurement of intracellular levels of free sphingoid bases.

Sphingoid bases are important for diverse cellular processes. This paper describes the development of a simple and rapid enzymatic method to quantify mass levels of free long-chain sphingoid bases in cellular lipid extracts. The assay is based on the ability of sphingosine kinase from Swiss 3T3 fibroblasts to phosphorylate sphingoid bases, predominantly sphingosine. To quantitatively determine cellular levels of sphingosine or other long-chain sphingoid bases, it was necessary to solubilize cellular lipids by sonication and to generate standard curves in the presence of lipid extracts containing at least 50 nmol of phospholipids. The assay conditions were optimized to allow quantification of sphingosine over a broad range from 25 to 1,000 pmol. Using this method we were able to obtain reproducible measurements of free sphingoid bases in various cell types and to detect increases in intracellular sphingoid base levels after treatment with exogenous sphingosine or stimulation with fetal bovine serum.

3T3 Cells↗
PubMed1994

Sphingosine-1-phosphate, a novel second messenger involved in cell growth regulation and signal transduction, affects growth and invasiveness of human breast cancer cells.

This review will focus on the role of sphingosine and its phosphorylated derivative sphingosine-1-phosphate (SPP) in cell growth regulation and signal transduction. We will show that many of the effects attributed to sphingosine in quiescent Swiss 3T3 fibroblasts are mediated via its conversion to SPP. We propose that SPP has appropriate properties to function as an intracellular second messenger based on the following: it elicits diverse cellular responses; it is rapidly produced from sphingosine by a specific kinase and rapidly degraded by a specific lyase; its concentration is low in quiescent cells but increases rapidly and transiently in response to the growth factors, fetal calf serum (FCS) and platelet derived growth factor (PDGF); it releases Ca2+ from internal sources in an InsP3-independent manner; and finally, it may link sphingolipid signaling pathways to cellular ras-mediated signaling pathways by elevating phosphatidic acid levels. The effects of this novel second messenger on growth, differentiation and invasion of human breast cancer cells will be discussed.

3T3 Cells↗
PubMed1994

The distribution of BrK alpha/RbK alpha peak intensity ratio in human whole blood samples. Comparison between normal and colorectal cancer cases.

A preliminary study was conducted to compare the BrK alpha/RbK alpha peak intensity ratio in colorectal cancer (CRC) patients and healthy subjects (controls) in Israel. Blood samples were obtained from 61 CRC patients and 124 controls. The controls represent a normal population from different areas in Israel. Three hundred microliters of wet whole blood samples were analyzed by the EDXRF method. A weighted mean of 2.45 +/- 0.38 Br/Rb ratio was obtained for CRC patients, as compared with 3.28 +/- 0.40 Br/Rb ratio for controls. The mean value for controls represents only 112 subjects, since 12 individuals of the control group suffered from some other diseases and therefore were not included for the mean value calculation in this group. The results indicate that the Br/Rb ratio in CRC patients is significantly lower (p < 0.05) than in controls. No significant difference was found in Br/Rb ratio correlated to age and sex. It was not possible to draw final conclusion concerning the relationship between the Br/Rb ratio and the malignancy stage, since there was a relatively small number of cases in each stage under investigation.

Adult↗
PubMed1994

Endogenous ganglioside GM1 modulates L-type calcium channel activity in N18 neuroblastoma cells.

Digital imaging fluorescence microscopy was used to investigate the effect of the B subunit of cholera toxin on calcium homeostasis in neuroblastoma N18 cells. The B subunit, which binds specifically to ganglioside GM1 in the outer leaflet of the cell membrane, was found to induce a sustained increase of intracellular calcium concentration ([Ca2+]i). The increase in [Ca2+]i was not observed in the absence of extracellular calcium, or in the presence of the calcium chelator EGTA, and was blocked by nickel. The B subunit was also found to induce an influx of manganese ions, as indicated by a quench of the intracellular fura-2 fluorescence. These data suggest that the B subunit induces an increase in calcium influx in N18 cells. Potassium-induced depolarization also stimulated manganese influx; however, after the onset of depolarization-induced influx, the B subunit had no further effect. This occlusion suggests involvement of voltage-dependent calcium channels. Treatment with BayK8644, a dihydropyridine agonist selective for L-type calcium channels, induced manganese influx that was not altered by the B subunit and apparently blocked the effect of the B subunit itself. Furthermore, the dihydropyridine L-type channel antagonists niguldipine or nicardipine completely inhibited B subunit-induced manganese influx. Thus, the B subunit-induced manganese influx is likely due to activation of an L-type voltage-dependent calcium channel. Spontaneous influx of manganese ions was also inhibited by nicardipine or niguldipine and by exogenous gangliosides. Ganglioside GM1 was more potent than GM3, but globoside had no significant effect. The modulation of L-type calcium channels by endogenous ganglioside GM1 has important implications for its role in neural development, differentiation, and regeneration and also for its potential function in the electrical excitability of neurons.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗
PubMed1993

Sphingosine-1-phosphate as second messenger in cell proliferation induced by PDGF and FCS mitogens.

Growth signalling networks that use glycerophospholipid metabolites as second messengers have been well characterized, but less is known of the second messengers derived from sphingolipids, another major class of membrane lipids. A tantalizing link between sphingolipids and cellular proliferation has emerged from the discovery that the sphingolipid metabolites sphingosine and sphingosine-1-phosphate stimulate growth of quiescent Swiss 3T3 fibroblasts by a pathway that is independent of protein kinase C. Sphingosine-1-phosphate is rapidly produced from sphingosine and may mediate its biological effects. Furthermore, sphingosine-1-phosphate triggers the dual signal transduction pathways of calcium mobilization and activation of phospholipase D, prominent events in the control of cellular proliferation. Here we report that activation of sphingosine kinase and the formation of sphingosine-1-phosphate are important in the signal transduction pathways activated by the potent mitogens platelet-derived growth factor (PDGF) and fetal calf serum (FCS).

3T3 Cells↗
PubMed1993

Signaling pathways for sphingosylphosphorylcholine-mediated mitogenesis in Swiss 3T3 fibroblasts.

Sphingosylphosphorylcholine (SPC), or lysophingomyelin, a wide-spectrum growth promoting agent for a variety of cell types (Desai, N. N., and S. Spiegel. 1991. Biochem. Biophys. Res. Comm. 181: 361-366), stimulates cellular proliferation of quiescent Swiss 3T3 fibroblasts to a greater extent than other known growth factors or than the structurally related molecules, sphingosine and sphingosine-1-phosphate. SPC potentiated the mitogenic effect of an activator of protein kinase C, 12-O-tetradecanoylphorbol 13-acetate, and did not compete with phorbol esters for binding to protein kinase C in intact Swiss 3T3 fibroblasts. However, downregulation of protein kinase C, by prolonged treatment with phorbol ester, reduced, but did not eliminate, the ability of SPC to stimulate DNA synthesis, indicating that SPC may act via both protein kinase C-dependent and -independent signaling pathways. SPC induced a rapid rise in intracellular free calcium ([Ca2+]i) in viable 3T3 fibroblasts determined with a digital imaging system. Although the increases in [Ca2+]i were observed even in the absence of calcium in the external medium, no increase in the levels of inositol phosphates could be detected in response to mitogenic concentrations of SPC. Furthermore, in contrast to sphingosine or sphingosine-1-phosphate, the mitogenic effect of SPC was not accompanied by increases in phosphatidic acid levels or changes in cAMP levels. SPC, but not sphingosine or sphingosine-1-phosphate, stimulates the release of arachidonic acid. Therefore, the ability of SPC to act an extremely potent mitogen may be due to activation of signaling pathway(s) distinct from those used by sphingosine or sphingosine-1-phosphate.

3T3 Cells↗
PubMed1993

Sphingosine and sphingosine 1-phosphate in cellular proliferation: relationship with protein kinase C and phosphatidic acid.

Previous studies from our laboratory have shown that sphingosine (Zhang et al. (1990) J. Biol. Chem. 265, 76-81) and sphingosine 1-phosphate (Zhang et al. (1991) J. Cell. Biol. 114, 155-167), metabolites of membrane sphingolipids, stimulate release of calcium from internal sources and increase proliferation of quiescent Swiss 3T3 fibroblasts acting in a fundamentally different, protein kinase C-independent pathway. The mitogenic effect of sphingosine was accompanied by an increase in the levels of phosphatidic acid (PA), a potent mitogen for a variety of cell types, that may function as an intracellular second messenger (Zhang et al. (1990) J. Biol. Chem. 265, 21309-21316). Sphingosine also induced early increases in sphingosine 1-phosphate (SPP) levels that preceded the increase in PA (Desai et al. (1992) J. Biol. Chem. 267, 23122-23128). SPP itself produced a more rapid increase in PA, thus suggesting that it may mediate the effects of sphingosine on PA accumulation. The concentration dependence for the formation of PA induced by SPP correlated with its effect on DNA synthesis. Similar to sphingosine, SPP also stimulated the activity of phospholipase D, although a significant effect was observed at a much lower concentration. However, in contrast to previous reports with sphingosine, SPP did not inhibit the PA phosphohydrolase activity in cell homogenates. Thus, in addition to its effect on mobilization of calcium, SPP can increase the level of PA, most likely via activation of phospholipase D. We suggest that SPP mediates the effect of sphingosine on PA accumulation in Swiss 3T3 fibroblasts and may regulate cellular proliferation by affecting multiple transmembrane signaling pathways.

Animals↗