Biomedical subjects

M Pierre

Publications and source records attributed to M Pierre.

At least 37 records · Page 2Linked to original sources

PubMed1997

Mediation by arachidonic acid metabolites of the H2O2-induced stimulation of mitogen-activated protein kinases (extracellular-signal-regulated kinase and c-Jun NH2-terminal kinase).

Reactive oxygen species modulate major cellular functions by mechanisms which are still poorly understood. Recently, H2O2 has been reported to stimulate the activity of the mitogen-activated protein kinases (MAPKs) ERK and JNK, and the expression of the proto-oncogenes c-fos and c-jun. As their expression is enhanced by H2O2 in astrocytes, we studied whether these MAPKs were stimulated by H2O2 in primary cultured astrocytes. The result was positive, a maximum of stimulation being reached with 200 microM H2O2 (0.3 pmol H2O2/cell) for both ERK and JNK. ERK was previously reported to stimulate cytosolic phospholipase A2 phosphorylation and activity. H2O2 stimulated the release of arachidonic acid in astrocytes, as already reported in other cell types. We found also that cPLA2 phosphorylation was increased by H2O2. Moreover, the stimulation by H2O2 of ERK and JNK was decreased by phospholipase A2 activity inhibitors. When astrocytes were incubated first with eicosatetraynoic acid, a structural analogue competing in arachidonic acid metabolism, the stimulation of JNK by H2O was also inhibited, suggesting the involvement of arachidonic acid metabolites. Cyclooxygenase or cytochrome P450 monooxygenase inhibitors failed in decreasing the MAPK stimulation by H2O2, whereas lipoxygenase inhibitors completely abolished that of JNK. Mitogenicity has been reported to be stimulated by H2O2 in other cell types. Although ERK was strongly and durably stimulated by 200 microM H2O2 in astrocytes, at the same extent as by mitogenic growth factors, basal thymidine incorporation rate was decreased by more than 80% after 12-15 h. Moreover, the stimulation of thymidine incorporation induced by basic fibroblast growth factor was transiently abolished by H2O2. Furthermore, H2O2 likely induced the expression of CL100/PAC1/MKP-1, a dual specificity phosphatase which has been implicated in ERK and JNK inactivation in the nucleus. Finally, the prior treatment of astrocytes with MK886, a 5-lipoxygenase-activating protein inhibitor, prevented JNK from stimulation, but did not prevent thymidine incorporation from inhibition, both induced by H2O2. These results strongly suggest an involvement of arachidonic acid and/or its metabolites in the stimulation of both ERK and JNK following the oxidative stress evoked by H2O2, which induced a cell cycle arrest probably independent of the stimulation of JNK.

Animals↗

PubMed1996

The Ras-GTPase-activating protein SH3 domain is required for Cdc2 activation and mos induction by oncogenic Ras in Xenopus oocytes independently of mitogen-activated protein kinase activation.

The Ras-GTPase-activating protein (RasGAP) is an important modulator of p21ras - dependent signal transduction in Xenopus oocytes and in mammalian cells. We investigated the role of the RasGAP SH3 domain in signal transduction with a monoclonal antibody against the SH3 domain of RasGaP. This antibody prevented the activation of the maturation-promoting factor complex (cyclin B-p34cdc2) by oncogenic Ras. The antibody appears to be specific because as little as 5 ng injected per oocyte reduced the level of Cdc2 activation by 50% whereas 100 ng of nonspecific immunoglobulin G did not affect Cdc2 activation. The antibody blocked the Cdc2 activation induced by oncogenic Ras but not that induced by progesterone, which acts independently of Ras. A peptide corresponding to positions 317 to 326 of a sequence in the SH3 domain of human RasGAP blocked Cdc2 activation, whereas a peptide corresponding to positions 273 to 305 of a sequence in the N-terminal moiety of the SH3 domain of RasGAP had no effect. The antibody did not block the mitogen-activated protein (MAP) kinase cascade (activation of MAPK/ERK kinase [MEK], MAP kinase, and S6 kinase p90rsk). Surprisingly, injection of the negative MAP kinase mutant protein ERK2 K52R (containing a K-to-R mutation at position 52) blocked the Cdc2 activation induced by oncogenic Ras as well as blocking the activation of MAP kinase. Thus, MAP kinase is also implicated in the regulation of Cdc2 activity. In this study, we further investigated the regulation of the synthesis of the c-mos oncogene product, which is necessary for the activation of Cdc2. We report that the synthesis of the c-mos oncogene product, which is necessary for the activation antibody to the SH3 domain of RasGAP and by injecting the negative MAP kinase mutant protein ERK2 K52R. These results suggest that oncogenic Ras activates two signaling mechanisms: the MAP kinase cascade and a signaling pathway implicating the SH3 domain of RasGAP. These mechanisms might control Mos protein expression implicated in Cdc2 activation.

Animals↗

PubMed1996

Evidence that type III iodothyronine deiodinase in rat astrocyte is a selenoprotein.

A type III iodothyronine deiodinase (D-III) that inactivates thyroid hormones has been recently cloned and identified as a selenoprotein in neonatal rat skin. However, selenium (Se) deficiency does not affect the D-III activity in the rat placenta and decreases the D-III in the rat brain only slightly. This study examines the effect of Se on the D-III activity in cultures of rat brain astrocytes. Astrocytes were depleted in Se by maintaining them in Se-free chemically defined medium for 7 days. These conditions decreased the activity of a recognized selenoprotein, glutathione peroxidase, 3-10-fold. D-III activity induced by 12-0-tetradecanoylphorbol-13-acetate (TPA) was also decreased 2-6-fold. Addition of 30 nM Se to the culture medium caused a rapid increase in TPA-induced D-III activity visible within 1 h. This Se effect was maximal at 3 h (4-fold increase) and dose-dependent. Se also increased the induction of D-III by acidic Fibroblast Growth Factor, 8-bromo-cAMP, T4, or retinoic acid. Cycloheximide blocked the effect of Se on TPA-induced D-III activity, whereas actinomycin D did not. Thus the rapid effect of Se does not require messenger RNA synthesis but requires protein synthesis. We conclude that the D-III in astrocytes is probably a selenoprotein.

8-Bromo Cyclic Adenosine Monophosphate↗

PubMed1995

Effects of growth factors on phosphatidylinositol-3 kinase in astroglial cells.

Growth factors differently regulate astroglial cell differentiation and proliferation. In an effort to understand the early intracellular events promoted by growth factors in astroglial cells, we have determined the effects of insulin-like growth factor I (IGF1), insulin, platelet-derived growth factor (PDGF), epidermal growth factor (EGF) and fibroblast growth factors (FGFs) on phosphatidylinositol-3 kinase (PI(3)-kinase). In astroglial cells cultured in serum-free medium, IGF1, PDGF, and EGF, which stimulate cell proliferation, increased PI(3)-kinase activity immunoprecipitated with anti-phosphotyrosine antibodies as shown by thin layer chromatography and high performance liquid chromatography. FGFa and FGFb, which strongly stimulate proliferation, glutamine synthetase, and deiodinase activities and modify cell morphology, have no effect on PI(3)-kinase activity. Addition of 1 nM PDGF, 10 nM IGF1, or 100 nM EGF to the culture medium rapidly stimulated PI(3)-kinase activity which declined slowly after 2 min. The stimulation of PI(3)-kinase increased with growth factor concentration. The maximum increase in PI(3)-kinase activity occurred with 50 nM IGF1, 1 nM PDGF, or 100 nM EGF. Since insulin was active only at high concentration (1 microM), its effect was probably mediated through IGF1 receptors and not through insulin receptors. IGF1 and PDGF, to a lesser degree, also increased the PI(3)-kinase activity associated with pp60c-src protein. Immunoblots performed with an antibody directed against the p85-subunit of the PI(3)-kinase confirmed that IGF1 increased the number of PI(3)-kinase molecules associated with phosphotyrosine-containing proteins or with c-src protein. Each growth factor affects in a different manner the association of PI(3)-kinase with phosphotyrosine-containing proteins and with pp60c-src and thus probably modulates intracellular signals downstream of PI(3)-kinase in astroglial cells.

Animals↗

PubMed1995

Stimulation of mitogen-activated protein kinase by thyrotropin in primary cultured human thyroid follicles.

In the thyroid, thyrotropin (TSH) stimulates both growth and function, and stimulates the production of cAMP which reproduces most of the effects of TSH. Here, we report evidence that TSH stimulates the mitogen-activated protein (MAP) kinase cascade through a cAMP-independent pathway, in human thyroid. TSH stimulated MAP kinase activity (4-9-fold the basal level) measured in the cytosolic fractions of primary cultured thyroid follicles. Maximal activity was reached after 20 min and remained sustained for 1-3 h, TSH being as potent as EGF; EC50 was 1.5 nM TSH. Only a single isoform of MAP kinase (p42) was detected in the follicles. p42 was phosphorylated on tyrosine residues and showed a reduced electrophoretic mobility in follicles stimulated by TSH. All these effects on MAP kinase were decreased by preincubation of the follicles with human anti-TSH receptor antibodies. The stimulation of MAP kinase by TSH was neither blocked by pertussis toxin nor reproduced by forskolin, cholera toxin, or 8-bromo-cAMP. In conclusion, in human thyroid cells, in contrast with previous observations on dog thyroid cells, TSH stimulates strongly MAP kinase through a pertussis toxin-insensitive and cAMP-independent pathway.

Amino Acid Sequence↗

PubMed1995

Stimulation of mitogen-activated protein kinase by thyrotropin in astrocytes.

We have recently reported the expression of the thyrotropin (TSH) receptor and the stimulation by TSH of type-II iodothyronine 5'-deiodinase in astrocytes. In these cells, TSH stimulated arachidonate release, but neither cAMP production, nor phosphatidylinositolbisphosphate hydrolysis, as described in the human thyroid gland. Here we report, in contrast to a recent observation made in dog thyroid cells, that TSH stimulates mitogen-activated protein kinase (MAP kinase) in astrocytes. Indeed, TSH increases the tyrosine phosphorylation of the two isoforms of MAP kinase expressed in these cells, in correlation with both a slower electrophoretic migration of the tyrosine phosphorylated species and an enhanced enzymic activity measured on a specific substrate peptide. This stimulation of MAP kinase by TSH was specifically inhibited by incubation of astrocytes in the presence of human blocking anti-(TSH receptor) IgG, and by immunoprecipitation of TSH with monoclonal anti-TSH IgG. In astrocytes, TSH was neither mitogenic by itself, nor modified significantly the basic-fibroblast-growth-factor-induced mitogenesis. The stimulation of MAP kinase by TSH was not affected by treatment with pertussis toxin, suggesting guanine-nucleotide-binding-regulatory protein i/o was not implicated in this TSH effect. Our model will allow the study of the stimulation of MAP kinase by TSH without interference either from cAMP or from phosphoinositide signalling pathways.

Animals↗

PubMed1995

Rapid TGF beta 1 effects on actin cytoskeleton of astrocytes: comparison with other factors and implications for cell motility.

We have previously shown that long-term treatment of primary cultured astrocytes with TGF beta 1 induces morphological changes accompanied by increases in actin and GFAP synthesis, and a profound rearrangement of the cytoskeleton. The present report describes the short-term reorganization of actin filaments induced by TGF beta 1 in rat cerebellum cultured astrocytes and in an astrocytic cell line. TGF beta 1 caused the appearance of new actin and vinculin organizations, without protein synthesis. This cytoskeletal rearrangement was followed by altered cell-cell interactions. All these changes induced by TGF beta 1 were different and slower than those induced by serum, PDGF, and endothelin. TGF beta 1 induced the appearance of lamellipodia, organelles found at the cell front of motile cells in low-density cultures of immortalized astrocytes. These results indicate that the changes in the astrocyte cytoskeleton induced by TGF beta 1 are probably associated with cell movement. The events promoted by TGF beta 1 might help to clarify its action in the brain during embryogenesis and in tissue repair.

Actins↗

PubMed1995

Improved training techniques and UltraBag system resulted in lowered peritonitis rate in an inner-city population.

Fifteen continuous ambulatory peritoneal dialysis (CAPD) patients from our dialysis unit were studied while using two different types of exchange systems: a snap-disconnect Y-set system and the Baxter UltraBag system. The incidence of peritonitis in the snap-disconnect Y-system was found to be 32 cases in 187 months, or a rate of 1 case of peritonitis every 5.8 months. When these patients switched to the Baxter UltraBag system, the incidence of peritonitis dropped to 4 cases in 175 months, or 1 case every 43.8 months, a highly significant difference. We conclude, therefore, that the UltraBag system is highly effective in reducing the incidence of peritonitis in CAPD patients.

Adolescent↗

PubMed1994

Inhibition of G1 cyclin expression and G1 cyclin-dependent protein kinases by cAMP in an astrocytic cell line.

The effects of cAMP on cell cycle progression were examined using an astrocytic cell line. We show that forskolin and 8-bromo-cAMP block the basic Fibroblast Growth Factor-induced DNA synthesis, do not inhibit mitogen activated protein kinase activation whereas they reduce G1 cyclin (E and D1) expression without modification of cyclin A level. Furthermore, they inhibit the activation of cyclin A- and cyclin E-dependent histone H1 kinases. These results suggest that cAMP may exert its antiproliferative effects through the regulation of cyclin synthesis and cyclin-dependent kinase activation.

Astrocytes↗

PubMed1994

Activation of the mitogen-activated protein kinase cascade by tyrphostin (RG 50864).

Tyrphostins are synthetic compounds which have been described as in vitro inhibitors of epidermal growth factor (EGF)-receptor tyrosine kinase activity. In NIH3T3 cells, stimulation of EGF-receptor tyrosine kinase leads to an increase of intracellular protein phosphorylations, among them the phosphorylation of mitogen-activated protein (MAP) kinase and the S6 kinases p90rsk and p70S6K. Phosphorylation of these proteins, either on tyrosine or serine/threonine residues or on both residues increases their protein kinase activity. Unexpectedly, treatment of NIH3T3 cells with both tyrphostin (RG 50864) and EGF results in an increase in the level of tyrosine phosphorylation of the MAP kinase. During this treatment, we also observed an increase in MAP kinase and S6 kinase p90rsk activities. Tyrphostin treatment diminishes the level of c-fos mRNA but has no effect on c-myc mRNA expression nor on S6 kinase p70S6K activity. Mitogenic signalling induced by EGF in NIH3T3 cells was blocked by tyrphostin, suggesting that the target(s) for this event may be elements downstream from the MAP kinase or independent of this signal transduction.

3T3 Cells↗

PubMed1994

Activation of the cell cycle machinery and the isoflavonoid biosynthesis pathway by active Rhizobium meliloti Nod signal molecules in Medicago microcallus suspensions.

We have shown that treatment of Medicago microcallus suspensions with the cognate Rhizobium meliloti Nod signal molecule NodRm-IV(C16:2,S) can modify gene expression both qualitatively and quantitatively. At concentrations of 10(-6) - 10(-9) M, this host specific plant morphogen but not the inactive non-sulfated molecule stimulated cell cycle progression as indicated by the significantly enhanced thymidine incorporation, elevated number of S phase cells, increase in kinase activity of the p34cdc2-related complexes and enhancement of the level of expression of several cell cycle marker genes, the histone H3-1, the cdc2Ms and the cyclin cycMs2. The presented data suggest that at least part of the physiological role of the Nod factor may be linked to molecular events involved in the control of the plant cell division cycle. In situ hybridization experiments with antisense H3-1 RNA probe indicated that only certain cells of the calli were able to respond to the Nod factor. High (10(-6) M) but not low (10(-9) M) concentrations of the active Nod factors induced the expression of the isoflavone reductase gene (IFR), a marker gene of the isoflavonoid biosynthesis pathway in most callus cells. Our results indicate that Medicago cell responses to the Nod signal molecules can be investigated in suspension cultures.

CDC2 Protein Kinase↗

PubMed1994

MAP kinase cascade in astrocytes.

We have studied in cultured rat astroglial cells MAP kinases, known for their role in intracellular signal transduction. The MAP kinase activity was stimulated by growth factors (FGFb, FGFa, EGF, PDGF, and IGF1), by a phorbol ester (TPA) activating-protein kinase C (PKC), by a neuropeptide (endothelin-1), and by a neuromediator (carbachol). Astrocytes pretreated for 18 h with TPA were still stimulated by growth factors and endothelin, suggesting that down-regulated isoforms of PKC are not involved in MAP kinase activation. In contrast, the small effect of carbachol was suppressed by TPA pretreatment. Astrocytes contained two proteins (p41 and p44) recognized by MAP kinase antibody. These proteins were phosphorylated on tyrosine residues in the cytosols of stimulated astrocytes. The kinetics of MAP kinase activation by FGFb and IGF1 were very different. FGFb promoted a rapid activation of MAP kinase (about 10 min) plus a prolonged phase that lasted at least 12 h. IGF1 produced only a rapid transient peak of activation at about 20 min. Hence, extracellular signals might generate different effects in astrocytes by differentially modulating the MAP kinase cascade. On a Mono Q column the growth factor-stimulated MAP kinase activity was separated into two peaks containing p41 and p44. Stimulation of astrocytes altered the elution pattern of p44 as a result of its phosphorylation. An ATP-dependent MAP kinase activator (MW = 40-45 kDa) was found in fractions of FGFb-stimulated cells which were not retained on Mono Q column, indicating the existence of a MAP kinase kinase (MEK) in astrocytes. C-Raf, identified in other cells as a MAP kinase kinase kinase, was also present in astrocytes.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1994

Induction of type III-deiodinase activity in astroglial cells by retinoids.

Thyroid hormones and retinoic acid (RA) are important modulators of growth, development, and differentiation. Type III deiodinase (D-III), which catalyzes thyroid hormones degradation in the brain and in cultured astroglial cells, is induced in astroglial cells by multiple pathways, including cAMP, 12.0-tetradecanoylphorbol-13-acetate (TPA), fibroblast growth factors, and thyroid hormones themselves. In the present study, the effects of retinoids on D-III activity were examined in astroglial cells cultures in a chemically defined medium devoid of hormones and growth factors. Incubation of astroglial cells with 5 microM all-trans-RA caused up to 200-fold increase in D-III activity, which reached a plateau after 48 h. The retinoid-induced increase in D-III activity was concentration dependent (0.5 microM all-trans-RA and 9-cis-RA producing half-maximal effect). Retinol was effective at physiological concentrations (1 and 10 microM). The 48 h effects of 5 microM all-trans-RA and 10 nM thyroid hormones on D-III activity were at least additive. Addition of 2 nM acidic fibroblast growth factor or 1 mM 8-bromo-cAMP for the last 8 h of a 48 h incubation with 5 microM all-trans-RA did not alter the induction by all-trans-RA, whereas 0.1 microM TPA in the same conditions produced an additive effect with all-trans-RA. All-trans-RA (5 microM) had little or no effect on type II deiodinase, the enzyme which catalyzes the activation of thyroxine to 3,5,3'-triiodothyronine.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1994

12-O-tetradecanoylphorbol 13-acetate and fibroblast growth factor increase the 30-kDa substrate binding subunit of type II deiodinase in astrocytes.

Type II 5'-deiodinase (D-II) catalyzes the intracellular conversion of thyroxine (T4) to 3,5,3'-triiodothyronine (T3) in the brain. The D-II activity in astroglial cell cultures is induced by several pathways including cyclic AMP (cAMP), 12-O-tetradecanoylphorbol 13-acetate (TPA), and fibroblast growth factors (FGFs). We have examined the effect of TPA and FGFs on the 30-kDa substrate binding subunit of D-II, by affinity labeling with N-bromoacetyl-[125I]T4 in astroglial cells. TPA (0.1 microM), 20 ng/ml acidic FGF (aFGF), and 1 mM 8-bromo cyclic AMP all caused an increase in the 30-kDa protein. cAMP induced the greatest increase (fivefold) followed by TPA (3.2-fold) and FGF (2.8-fold). Glucocorticoids acted synergistically with cAMP and aFGF and promoted the effect of TPA. Affinity labeling was competitively inhibited by bromoacetyl-T4 > bromoacetyl-T3 > T4 > reverse T3 > iopanoic acid > T3 > 3,5,3'-triiodothyroacetic acid. The effect of TPA (0.1 microM) was maximum at 8 h and then gradually decreased. aFGF (20 ng/ml) plus heparin (17 micrograms/ml) induced a maximal 30-kDa increase at 8 h, which stayed stable for up to 24 h. The effect of aFGF was concentration dependent. Of the other growth factors studied, only basic FGF and platelet-derived growth factor induced small increases in the 30-kDa protein. Epidermal growth factor had little effect. In vitro labeling of cAMP, TPA, and aFGF-stimulated cell sonicates resulted in an increase in the 30-kDa protein that paralleled the increase in D-II activity.(ABSTRACT TRUNCATED AT 250 WORDS)

8-Bromo Cyclic Adenosine Monophosphate↗

PubMed1994

Sulfation after deiodination of 3,5,3'-triiodothyronine in rat cultured astrocytes.

The metabolism of [125I]T3 by rat astrocytes in culture was analyzed by Sephadex LH-20 chromatography and HPLC. The conjugates isolated on LH-20 were not hydrolyzed by glucuronidase, indicating the absence of glucuroconjugates. 3,3'-Diiodothyronine (3,3'T2) sulfate (3,3'T2-S) was the main product that accumulated in the medium over the T3 concentration explored (10 pM to 10 nM). The identity of the peak eluted as 3,3'T2-S was ascertained by its hydrolysis with sulfatase and the generation of 3,3'T2 identified by HPLC. 3'-Monoiodothyronine sulfate was also found in cells treated with 1 microM retinoic acid, i.e. with high type III deiodinase activity. No T3 sulfate (T3-S) was found as a metabolite of T3. Astrocytes did not break down 1 nM [125I]T3-S added to the medium. Astrocytes pretreated for 3 days with 10 nM T3 showed increased production of 3,3'T2-S from 10 nM [125I]T3. Exogenous [125I]3,3'T2 (20 nM) was conjugated to 3,3'T2-S released into the medium. Pretreatment of astrocytes with 10 nM T3 did not alter the production of 3,3'T2-S from 3,3'T2. Thus, T3 is metabolized in astrocytes by direct 5-deiodination, followed by sulfation. Whereas T3 induces its own deiodination and type III deiodinase activity, T3 does not regulate the sulfation of its main metabolite, 3,3'T2. This demonstration of sulfation of iodothyronines in cells originating from the brain raises the question of the role of this TH metabolic pathway in the brain.

Animals↗

PubMed1994

Growth factor-regulated phosphatidylinositol-3-kinase in astrocytes. Involvement of pp60c-src.

Growth factors differently regulate astroglial cell differentiation and proliferation. In an effort to understand the early intracellular events promoted by growth factors in astroglial cells, we have determined the effects of IGF1, insulin, PDGF, EGF and FGFs on phosphatidylinositol-3 kinase. IGF1, PDGF and EGF which stimulate cell proliferation of astroglial cells, increased phosphatidylinositol-3 kinase activity immunoprecipitated with anti-phosphotyrosine antibodies as shown by thin layer chromatography and high performance liquid chromatography. FGFa and FGFb, which strongly stimulate proliferation of astroglial cells, have no effect on phosphatidylinositol-3 kinase activity. Addition of 1 nM PDGF, 10 nM IGF1 or 100 nM EGF to the culture medium rapidly stimulated phosphatidylinositol-3 kinase activity which declined slowly after 2 min. The stimulation of phosphatidylinositol-3 kinase increased with growth factor concentration. The maximum increase in phosphatidylinositol-3 kinase activity occurred with 50 nM IGF1, 1 nM PDGF or 100 nM EGF. Since insulin was active only at high concentration (1 microM), its effect was probably mediated through IGF1 receptors and not through insulin receptors. Treatment with IGF1-plus-EGF, had an additive effect on PI(3)-kinase activity, PDGF-plus-IGF1 did not. IGF1 and PDGF, to a lesser degree, also increased the phosphatidylinositol-3 kinase activity associated with pp60c-src protein. Immunoblots performed with an antibody directed against the p85-subunit of the phosphatidylinositol-3 kinase confirmed that IGF1 increased the number of phosphatidylinositol-3 kinase molecules associated with phosphotyrosine-containing proteins or with c-src protein. Each growth factor affects in a different manner the association of phosphatidylinositol-3 kinase with phosphotyrosine-containing proteins and with c-src protein and thus probably modulates intracellular signals downstream of phosphatidylinositol-3 kinase in astroglial cells.

Animals↗

PubMed1993

Evidence for cAMP-independent thyrotropin effects on astroglial cells.

Thyroid hormones are essential for normal brain development and function. Brain astroglial cells express type II iodothyronine 5'-deiodinase which converts thyroxine into 3,5,3'-triiodothyronine. This type II deiodinase is regulated through various signalling pathways, allowing probably for the local adaptation of the level of 3,5,3'-triiodothyronine. Our results demonstrated that thyrotropin was able to induce type II deiodinase activity in astrocytes. A thyrotropin receptor was demonstrated. It was not coupled, as in thyroid, to adenylyl cyclase and phospholipase C, but it stimulated cytosolic phospholipase A2. The stimulation by thyrotropin of both thyroxine synthesis in thyroid and its local activation in astrocytes, could protect the brain from variations in the level of 3,5,3'-triiodothyronine.

Adenylyl Cyclases↗

PubMed1993

Effects of transforming growth factor-beta 1 on the extracellular matrix and cytoskeleton of cultured astrocytes.

The present study was performed on primary cultures and subcultures of cerebellar astrocytes in order to investigate the effects of transforming growth factor-beta 1 (TGF beta 1) on proliferation, extracellular matrix (ECM) components, and cytoskeletal structures in relation to morphological changes. The expression and cellular distribution of the ECM components laminin and fibronectin and the cytoskeletal proteins glial fibrillary acidic protein (GFAP) and actin were investigated by immunoblotting, immunocytochemistry, and phalloidin staining. The proliferation of primary cultures was strongly inhibited by TGF beta 1. Treated cells became enlarged and spread onto the substratum. TGF beta 1 promoted the appearance of actin stress fibers and increased the cell actin content. It elicited a slight increase in GFAP expression and induced dispersion of thin filaments of GFAP. TGF beta 1 also stimulated the production of laminin and fibronectin and their incorporation into the ECM of primary cultures grown in medium with or without serum. Astrocytes grown in serum-containing medium for 1 day after subculturing responded strongly to TGF beta 1. Changes promoted by TGF beta 1 in cell shape, cytoskeleton, and ECM production of cultured astrocytes may have relevance for understanding the mechanisms of action of TGF beta 1 during brain development.

Actins↗