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Biomedical subjects

B Fauconneau

Publications and source records attributed to B Fauconneau.

At least 19 recordsLinked to original sources

Plasmalogen degradation by oxidative stress: production and disappearance of specific fatty aldehydes and fatty alpha-hydroxyaldehydes.

Plasmalogens are often considered as antioxidant molecules that protect cells from oxidative stress. Their vinyl ether bond could indeed be among the first targets for newly formed radicals. However, the long chain aldehydes released from plasmalogens were seldom studied and possible injurious or harmless effects were poorly examined. Thus, the sensitivity of the vinyl ether bond of plasmalogens was investigated in a cerebral cortex homogenate under UV irradiation- or Fe2+/ascorbate-induced peroxidation. Kinetics of aldehyde production was followed by gas chromatography/mass spectrometry. This confirmed that plasmalogens were highly sensitive to oxidative stress (70% cleavage after 90 min UV irradiation and 30% after 30 min of Fe2+/ascorbate). The aldehydes corresponding to sn-1 position 16:0, 18:0, or 18:1 were poorly detected. Conversely, oxidation of plasmalogens yielded preferentially 15:0, 17:0, and 17:1 aldehydes under UV and the alpha-hydroxyaldehydes 16:0-OH and 18:0-OH following a Fe2+/ascorbate oxidation. Kinetics showed that free aldehydes and above all free alpha-hydroxyaldehydes disappeared from the medium as soon as produced. Consequently, the behavior of these released aldehydes in the tissues has to be investigated in order to ascertain the protective effect of plasmalogens against oxidation.

Aldehydes↗

Impairment of the neuronal dopamine transporter activity in MPP(+)-treated rat was not prevented by treatments with nitric oxide synthase or poly(ADP-ribose) polymerase inhibitors.

The neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) causes, via its metabolite MPP(+), damages of the nigrostriatal dopaminergic pathway, similar to those observed in Parkinson's disease. An intranigral injection of 10 microg MPP(+) in rat induced a decrease of about 30% of the neuronal dopamine transporter (DAT) activity 21 days after lesion. Based on the hypothesis that MPTP/MPP(+) neurotoxicity involves the nitric oxide (NO) production and/or an activation of poly(ADP-ribose) polymerase (PARP), we investigated the preventive effects of a treatment either with L-Name, a NO synthase (NOS) inhibitor or 3-aminobenzamide, a PARP inhibitor on the reduction of dopamine uptake induced by MPP(+). Rats received a daily injection i.p. of 50 mg/kg L-Name or 10 mg/kg 3-aminobenzamide 3 days before and during 21 days after the MPP(+) lesion. The results showed that inhibitors of NOS and PARP did not prevent the alteration of DAT activity induced by 10 microg MPP(+), indicating that NO and PARP were not involved in the biochemical cascade leading to the inhibition of rat DAT activity by MPP(+) in our experimental conditions.

1-Methyl-4-phenylpyridinium↗

Hypothyroidism induces type I iodothyronine deiodinase expression in tilapia liver.

In the current study, the authors examined the effects of experimentally induced hypothyroidism on peripheral thyroid hormone metabolism and growth in two closely related tilapia species: the Nile tilapia (Oreochromis niloticus) and the slower growing black tilapia (Sarotherodon melanotheron). Hypothyroidism, induced by administration of 0.2% methimazole through the food, significantly decreased plasma T(3) and T(4) in both species. This decrease in circulating thyroid hormones was accompanied by an increase in hepatic type II deiodinase (D2) and a decrease in hepatic type III deiodinase (D3). Hepatic type I deiodinase (D1), which is barely expressed in euthyroid tilapia, was significantly upregulated during hypothyroidism. The changes in hepatic D1 and D2 enzyme activity were paralleled by changes in D1 and D2 mRNA levels, indicating pretranslational regulation. Hypothyroidism also resulted in severe growth retardation that was accompanied by an increase in condition factor. Because hyperthyroidism has been shown to decrease the condition factor, these results suggest that thyroid hormones play an essential role in the control of proportional body growth in fish. The authors conclude that (1) hepatic D1 expression is induced by hypothyroidism in tilapia, (2) the changes in hepatic iodothyronine deiodinases during hypothyroidism in tilapia are predominantly regulated at a pretranslational level, and (3) thyroid hormones are involved in the control of proportional body growth in fish.

Amino Acid Sequence↗

Evidence against a major role of plasmalogens in the resistance of astrocytes in lactic acid-induced oxidative stress in vitro.

Astrocytes are known to play a key role in buffering extracellular pH variations and, in addition, they are particularly resistant to oxidative stress and subsequent lipid peroxidation. This great resistance may be ascribed to the presence of high concentrations of certain antioxidants, but another explanation may be the presence of a high quantity of plasmalogens, which are a special group of glycerophospholipids characterized by a vinyl ether bond instead of an ester bond in the sn-1 position of the glycerol backbone. Plasmalogens are sensitive to free radical attack and acidity, and numerous works have supported the hypothesis that they may be antioxidant molecules that protect cells from oxidative stress. The aim of this work was to investigate, on astrocytes in lactic acid-induced oxidative stress (pH 5.5), the behavior of phospholipids and, in particular, plasmalogens. Two main techniques, based on the susceptibility of the vinyl ether bond to hydrolysis, were employed in this study to measure plasmalogen levels. In both cases, the sn-1 vinyl ether linkage was cleaved using mercuric chloride, producing a lysophospholipid that was assessed by phosphorus measurement or using HCl treatment, producing a long-chain fatty aldehyde assayed by gas chromatography/mass spectrometry. On astrocytes in culture, only plasmenylethanolamine (PlmEtn) was evidenced, representing 40% of glycerophosphoethanolamine lipids. When astrocytes were incubated with lactic acid, no modification in the amount of PlmEtn was seen. Furthermore, free aldehydes and aldehydes corresponding to the quantity of intact plasmalogens were similar to those observed on controls. In addition, the constancy of two lipid peroxidation markers, thiobarbituric acid reactive substances and polyunsaturated fatty acids, was clear evidence of the resistance of these cells in lactic acid conditions. In conclusion, our results fail to demonstrate a major role of plasmalogens in the resistance of astrocytes in lactic acid-induced oxidative stress.

Acidosis, Lactic↗

Sensitivity of muscle satellite cells to pollutants: an in vitro and in vivo comparative approach.

Muscle satellite cells from rainbow trout were exposed in vitro to increasing concentrations of different xenobiotics: copper, dichloroaniline, prochloraz, nonyl-phenol polyethexylate. Mortality and proliferation rate were measured by Hoechst binding and BrdU incorporation. Dose dependent effect of copper on survival and proliferation was observed with an EC50 at 100 microM. A dose dependent effect of nonyl-phenol diethoxylate was observed on survival with an EC50 at 100 microM. This was associated with a biphasic effect on proliferation rate observed both for nonyl-phenol di and poly-ethoxylate: a stimulation of proliferation at low concentration and an inhibition proliferation at large concentration. These effects were related to the inhibition of cells adhesion through the detergent capacity of nonyl-phenol polyethoxylate. The effects of prochloraze and dichloroaniline on cells mortality (EC 50 > 500 microM) and proliferation rate (LOEC: 100 microM) were limited. Whole fish growth, muscle fibre size distribution and satellite cells survival and proliferation were measured on rainbow trout (60-80 g BW) exposed to two concentrations of prochloraze (10 and 100 microg/ml) or nonyl phenol diethoxylate IGEPAL 210 (100 and 400 microg/ml) during 14 and 10 days exposure, respectively. Muscle fibre size distribution and satellite cells proliferation were affected by prochloraz exposure in vivo and this could be related to the alteration in fish feeding status. The exposure to IGEPAL 210 affected the number of satellite cells extracted and induce a biphasic effect on satellite cells proliferation similar to that observed in vivo. The combined exposure to IGEPAL 210 and prochloraze revealed additive effects of these two compounds. The in vivo and in vitro comparison demonstrated that in vitro satellite cells system could be used as a valuable tool to test the effects of pollutants.

Aniline Compounds↗

A new in vitro approach for investigating the MPTP effect on DA uptake.

Previous studies have shown that dopamine (DA) uptake was decreased after preincubation of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) or 1-methyl-4-phenylpyridinium (MPP(+)) in in vitro slice and synaptosome models. The present study, conducted with and without preincubation, attempted to determine whether inhibition results from a direct effect of neurotoxins on neuronal DA transporter or from an alteration of the transporter secondary to other toxic events. DA uptake was inhibited about 50% in the presence of MPTP+O(2) or MPP(+) (0.1, 1 and 5 mM) in rat striatal slices and synaptosomes. Such inhibition was obtained in synaptosomes preincubated for 150 min with MPP(+) and then washed. Inhibition of DA uptake was lower in slices preincubated with MPTP (5 mM)+O(2) and then washed (30%). Experiments in synaptosomes prepared from slices preincubated with MPTP or MPP(+) showed greater inhibition of DA uptake with MPTP. The results suggest that the inhibition of DA uptake in vitro by MPTP or MPP(+) results initially from a direct effect on the transporter during its penetration in nerve endings and subsequently from a transporter alteration related to toxic events. Thus, the preincubation of striatal slices followed by DA uptake measurement in synaptosomes would appear to be a good in vitro model for studying the dopaminergic toxicity of MPTP.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗

Lower environmental temperature delays and prolongs myogenic regulatory factor expression and muscle differentiation in rainbow trout (Onchrhynchus mykiss) embryos.

The effect of different temperatures (4 degrees C and 12 degrees C) on myogenic regulatory factors (MyoD and myogenin) and myosin heavy chain (MyHC) expression was investigated in rainbow trout (Onchrhynchus mykiss) during early development. MyoD is first switched on at stage 14 [about 5 somites are formed (1/2 epiboly)] while myogenin mRNA is expressed at stage 15 [around 15 somites are visible (2/3 epiboly)] at both temperatures. Subsequently (up to at least stage 20), the most caudal somites exhibit less myogenin mRNA at 4 degrees C compared to 12 degrees C. At the eyed stage (stage 23-24), both myogenin mRNA and protein are present in greater amounts throughout all myotomes at the lower temperature, with mRNA levels in warmer (12 degrees C) embryos at 83% for MyoD and 72% for myogenin of the levels seen in 4 degrees C embryos. Conversely, however, at this same stage, fast-MyHC mRNA and protein are more abundant in 12 degrees C than in 4 degrees C embryos. This indicates relatively advanced muscle differentiation at the warmer temperature. At hatching, myogenin-positive cells are concentrated within the myosepta at both temperatures and they are also sparsely distributed in the myotome at 4 degrees C, but not at 12 degrees C. MyoD, myogenin, and MyHC levels provide an indication of differentiation of muscle cells. These findings suggest that myogenic regulatory factor expression is delayed but prolonged by the lowering of temperature.

Animals↗

Red and white muscle development in the trout (Oncorhynchus mykiss) as shown by in situ hybridisation of fast and slow myosin heavy chain transcripts.

The axial muscle of most teleost species consists of a deep bulk of fast-contracting white fibres and a superficial strip of slow-contracting red fibres. To investigate the embryological development of fast and slow muscle in trout embryos, we carried out single and double in situ hybridisation with fast and slow myosin heavy chain (MyHC)-isoform-specific riboprobes. This showed that the slow-MyHC-positive cells originate in a region of the somite close to the notochord. As the somite matures in a rostrocaudal progression, the slow-MyHC-positive cells appear to migrate radially away from the notochord to the lateral surface of the myotome, where they form the superficial strip of slow muscle. Surprisingly, the expression pattern of the fast MyHC showed that the differentiation of fast muscle commences in the medial domain of the somite before the differentiation and migration of the slow muscle precursors. Later, as the differentiation of fast muscle progressively spreads from the inside to the outside of the myotome, slow-MyHC-expressing cells become visible medially. Our observations that the initial differentiation of fast muscle takes place in proximity to axial structures and occurs before the differentiation and migration of slow muscle progenitors are not in accord with the pattern of muscle formation in teleosts previously described in the zebrafish Danio rerio, which is often used as the model organism in fishes.

Amino Acid Sequence↗

Temperature and myogenic factor transcript levels during early development determines muscle growth potential in rainbow trout (Oncorhynchus mykiss) and sea bass (Dicentrarchus labrax).

The influence of changes in environmental temperature on the mRNA levels of myogenic regulatory factors (MRFs), i.e. MyoD and myogenin, as well as myosin heavy chain (MyHC) were studied during early larval development in rainbow trout and sea bass. Phosphoimager analysis of northern blots indicated that there is an optimum temperature for the RNA transcript levels of MRF and MyHC RNA in trout and in sea bass larvae. In the trout strain studied, the highest concentration for MRF and MyHC transcripts was found at 8 degrees C rather than 4 degrees C or 20 degrees C. In European sea bass, the highest concentrations of MRF and MyHC mRNA were observed at 15-20 degrees C rather than 13 degrees C. Raising sea bass larvae at 15 degrees C was associated with higher MyHC gene expression as well as a trend towards an increase in total muscle fibre number and higher growth rates after transfer at ambient temperature. Results suggest that mRNA levels of MRF and MyHC can be used to optimise early development. An experiment in which the temperature was changed illustrates the consequence of precise temporal expression of MRF genes in specifying muscle fibre number at critical stages during early development.

Animals↗

Embryonic muscle development in rainbow trout (Oncorhynchus mykiss): a scanning electron microscopy and immunohistological study.

Embryonic muscle development was studied in rainbow trout (Oncorhynchus mykiss) at low and high temperature using scanning electron microscopy (SEM) and immunohistology. Somite development was described starting at stage 16 (Vernier JM. 1969. Ann Embryol Morphogen 4:495-520) for both temperatures, with special interest in their shape and size. Muscle differentiation, associated with somite growth, is characterized by a larger increase in height compared to width and by acquisition of a chevron shape. Thin structures such as striation, sarcomeres, and myofibrils within muscle cells and myotubes were observed starting at the eyed stage (stage 24). Immunohistological analyses showed appearance of embryonic fast myosin at stage 20 in the deep part of the somite. The area where myosin was expressed extended in the somite throughout embryonic development and the presence of myosin was observed in the entire somite at hatching (stage 30). Slow myosin was expressed in a monolayer of superficial cells at the eyed stage and during the entire embryonic development. Then it was expressed in a few layers of cells located in the red muscle area. These results suggest that muscle differentiation, characterized by myosin expression, is engaged at stage 20. Myogenesis starts in the deep part of the somite, near the notochord and progresses laterally to cover the complete somite at hatching when the somite is composed of muscle fibres exhibiting a high degree of maturity. No significant difference was observed in terms of muscular development between low- and high-temperature conditions. J. Exp. Zool. 286:379-389, 2000.

Animals↗

Effect of fasting and refeeding on in vitro muscle cell proliferation in rainbow trout (Oncorhynchus mykiss).

The effects of short-term fasting and refeeding were studied on satellite cells extracted from white epaxial muscle of juvenile rainbow trout (1-3 g body weight). In vitro changes in the proliferation of satellite cells were analyzed using bromodeoxyuridine (BrdU) incorporation over a 24-h period. Proliferation in fed control fish was characterized by an initial basal proliferation rate of 5-10% BrdU-labeled nuclei x day(-1), followed by an exponential increase at a rate of +18-20% x day(-1), up to a maximum of 60-70% BrdU-labeled nuclei x day(-1). Characteristics of satellite cells extracted from starved fish, namely extraction yield, morphology, and proliferation, were different from those of fed fish. Fasting (8-10 days) completely suppressed initial proliferation of satellite cells in vitro over a period of 4 days. After this delay, proliferation resumed and changes in proliferation rates over time were similar to those of the control group. In fish fed for 4 days after an 8-day fast, the initial proliferation rate and the changes in proliferation rates over time were completely restored. These findings demonstrate that satellite cells express different behavior depending on feeding status, which could be due to the presence of different satellite cell populations.

Age Factors↗

Origin of 4-hydroxynonenal incubation-induced inhibition of dopamine transporter and Na+/K+ adenosine triphosphate in rat striatal synaptosomes.

Previous experiments reported that an incubation of striatal synaptosomes with 4-hydroxynonenal (4-HNE) resulted in an inhibition of dopamine (DA) uptake and Na+/K+ adenosine triphosphate (ATPase) activity. The present work investigated whether theses inhibitions are related to a 4-HNE binding to the DA transporter (DAT) and the Na+/K+ ATPase. The number of specific [125I]-PE21 binding sites on the DAT was significantly reduced after incubation with 4-HNE. The Na+/K+ ATPase activity decrease induced by 4-HNE was partially reversed, in a dose-dependent manner, by veratridine, a pump stimulator agent. Our previous data (Morel, P., Tallineau, C., Pontcharraud, R., Piriou, A. and Huguet, F., Effects of 4-hydroxynonenal, a lipid peroxidation product, on dopamine transport and Na+/K+ ATPase in rat striatal synaptosomes. Neurochem. Int., 33 (1999) 531-540) combining with the data observed in this study suggest that changes in DA uptake in striatal synaptosomes are directly related to 4-HNE binding to the DAT, whereas the decrease in Na+/K+ ATPase activity resulted only partially from 4-HNE binding to the pump and is mainly secondary to membrane lipid disruption.

Adenosine Triphosphatases↗

Synthesis and antioxidant activity of new tetraarylpyrroles.

The synthesis and in vitro antioxidant activity of 17 new tetraarylpyrroles are investigated by 2 tests highly documented in the literature: capability to prevent Fe(2+)-induced lipid peroxidation on microsomes, which is a membrane preparation rich in polyunsaturated fatty acids, and direct scavenging effect on a stable free radical, 1,l-diphenyl-2-picryl-hydrazyl (DPPH). For the Fe(2+)-induced microsomal lipid peroxidation system, the results show that molecules which possess 2-pyrazinyl or 2-pyridyl in the 3- and 4-positions on the pyrrole ring are the most efficient. Introduction of methoxy groups on the phenyl ring in the 2- and 5-positions increases the effects but the higher activity is obtained with 2-furyl or 2-thienyl. The only compounds which possess a direct scavenger effect on trapping the stable free radical DPPH are those which have 2-pyridyl in the 3- and 4-positions and 2-furyl or 2-thienyl in the 2- and 5-positions.

Journal Article↗

Membrane carbohydrate conjugates desialylation does not alter [3H]-dopamine uptake in rat striatal slices.

Incubation of rat striatal slices induced a large decrease (about 50%) of DA uptake and a slight desialylation of polysialogangliosides (GT1b, GD1b, GD1a) with an increase of monosialogangliosides (GM1). Moreover, a pretreatment of slices by exogenous added neuraminidase of Vibrio cholerae did not modify DA uptake, although the pattern of gangliosides was modified and there was considerable loss (about 45%) of sialic acid in gangliosides and glycoproteins. It was verified that neuraminidase activity occured in synaptic membrane. Thus, DA uptake was apparently not altered by desialylation of plasma membrane carbohydrate conjugates.

Animals↗

Inhibitory effects of ascorbic acid on dopamine uptake by rat striatal synaptosomes: relationship to lipid peroxidation and oxidation of protein sulfhydryl groups.

Ascorbic acid is frequently added in the incubation medium to prevent oxidation of dopamine (DA) during uptake assays. However, a preliminary study showed that the presence of ascorbic acid induced a decrease of DA uptake after prolonged incubation. The purpose of this study was to determine the mechanism underlying ascorbic acid-induced alterations of DA uptake in rat striatal synaptosomes. In this context, the effects of physiological concentrations of ascorbic acid (100-500 microM) on DA uptake and Na+/K+ ATPase activity (which is essential for DA transporter function) were assessed in synaptosomes before and after incubation at 37 degrees C. The capacity of synaptosomes to take up DA was significantly decreased after incubation owing to a reduction in DA transporters (but with no modification of their affinity for DA). This partial inhibition was associated with a decrease of Na+/K+ ATPase activity, a production of thiobarbituric acid reactive substances (TBARS) and malonaldehyde (MDA), and a loss of sulfhydryl group content. Addition of Trolox C to the medium prevented the reduction of DA uptake, the inhibition of Na+/K+ ATPase activity, the decrease in sulfhydryl group content and the production of TBARS and MDA. These results suggest that ascorbic acid in the presence of contaminant ferrous ions induced a decrease in functional DA transporters, probably through a lipid peroxidation process involving oxidation of sulfhydryl groups and at least in part through a decrease of Na+/K+ ATPase activity.

Animals↗

Isolation, identification, and antioxidant activity of three stilbene glucosides newly extracted from vitis vinifera cell cultures

Suspension cultures of Vitis vinifera L. (Vitaceae) produce many hydroxylated stilbene glucosides found in red wine. From these cells, we isolated and characterized glycosylated stilbenes, (Z)-piceatannol (3,5,3',4'-tetrahydroxystilbene) -3-O-beta-d-glucopyranoside (6) and (E)- and (Z)-resveratrol (3,5, 4'-trihydroxystilbene)-4'-O-beta-d-glucopyranoside (2 and 7, respectively), which have not previously reported to be constituents of Vitis vinifera or wine. The ability of these compounds to act as radical scavengers was investigated using 1,1 diphenyl-2-picryl-hydrazyl, a stable free radical. Antioxidant activities were assessed by their capacity to prevent Cu2+-induced lipid peroxidation in human low-density lipoprotein.

Journal Article↗