GALNT2 as a novel modulator of adipogenesis and adipocyte insulin signaling

Background/objectives: A better understanding of adipose tissue biology is crucial to tackle insulin resistance and eventually coronary heart disease and diabetes, leading causes of morbidity and mortality worldwide. GALNT2, a GalNAc-transferase, positively modulates insulin signaling in human liver cells by down-regulating ENPP1, an insulin signaling inhibitor. GALNT2 expression is increased in adipose tissue of obese as compared to that of non-obese individuals. Whether this association is secondary to a GALNT2-insulin sensitizing effect exerted also in adipocytes is unknown. We then investigated in mouse 3T3-L1 adipocytes the GALNT2 effect on adipogenesis, insulin signaling and expression levels of both Enpp1 and 72 adipogenesis-related genes. Methods: Stable over-expressing GALNT2 and GFP preadipocytes (T 0 ) were generated. Adipogenesis was induced with (R+) or without (R−) rosiglitazone and investigated after 15 days (T 15 ). Lipid accumulation (by Oil Red-O staining) and intracellular triglycerides (by fluorimetric assay) were measured. Lipid droplets (LD) measures were analyzed at confocal microscope. Gene expression was assessed by RT-PCR and insulin-induced insulin receptor (IR), IRS1, JNK and AKT phosphorylation by Western blot. Results: Lipid accumulation, triglycerides and LD measures progressively increased from T 0 to T 15 R- and furthermore to T 15 R+. Such increases were significantly higher in GALNT2 than in GFP cells so that, as compared to T 15 R+GFP, T 15 R- GALNT2 cells showed similar (intracellular lipid and triglycerides accumulation) or even higher (LD measures, p < 0.01) values. In GALNT2 preadipocytes, insulin-induced IR, IRS1 and AKT activation was higher than that in GFP cells. GALNT2 effect was totally abolished during adipocyte maturation and completely reversed at late stage maturation. Such GALNT2 effect trajectory was paralleled by coordinated changes in the expression of Enpp1 and adipocyte-maturation key genes. Conclusions: GALNT2 is a novel modulator of adipogenesis and related cellular phenotypes, thus becoming a potential target for tackling the obesity epidemics and its devastating sequelae

Enhanced insulin sensitivity associated with provision of mono and polyunsaturated fatty acids in skeletal muscle cells involves counter modulation of PP2A

International audienceAims/Hypothesis: Reduced skeletal muscle insulin sensitivity is a feature associated with sustained exposure to excess saturated fatty acids (SFA), whereas mono and polyunsaturated fatty acids (MUFA and PUFA) not only improve insulin sensitivity but blunt SFA-induced insulin resistance. The mechanisms by which MUFAs and PUFAs institute these favourable changes remain unclear, but may involve stimulating insulin signalling by counter-modulation/repression of protein phosphatase 2A (PP2A). This study investigated the effects of oleic acid (OA; a MUFA), linoleic acid (LOA; a PUFA) and palmitate (PA; a SFA) in cultured myotubes and determined whether changes in insulin signalling can be attributed to PP2A regulation. Principal Findings: We treated cultured skeletal myotubes with unsaturated and saturated fatty acids and evaluated insulin signalling, phosphorylation and methylation status of the catalytic subunit of PP2A. Unlike PA, sustained incubation of rat or human myotubes with OA or LOA significantly enhanced Akt-and ERK1/2-directed insulin signalling. This was not due to heightened upstream IRS1 or PI3K signalling nor to changes in expression of proteins involved in proximal insulin signalling, but was associated with reduced dephosphorylation/inactivation of Akt and ERK1/2. Consistent with this, PA reduced PP2Ac demethylation and tyrosine 307 phosphorylation-events associated with PP2A activation. In contrast, OA and LOA strongly opposed these PA-induced changes in PP2Ac thus exerting a repressive effect on PP2A.Conclusions/Interpretation: Beneficial gains in insulin sensitivity and the ability of unsaturated fatty acids to oppose palmitate-induced insulin resistance in muscle cells may partly be accounted for by counter-modulation of PP2A

Distribution and History of Naturalized Common Pear, Pyrus communis, in Ontario

Considered rare or entirely overlooked until 1998, Pyrus communis is currently widespread and locally abundant as an escape from cultivation in southern Ontario. It was first noticed growing wild in Elgin County in 1949 and up until 1965 it was known as a wild plant only in the eastern Lake Erie region. It was first reported in the eastern Lake Ontario region in 1969 and is now widespread and locally abundant there. It now occurs north to the Georgian Bay region, and southwest to Essex County. Since it has had a very effective and rapid dispersal system for centuries as a result of cultivation and discarded cores, the apparently recent spread may be a consequence of climate warming extending the area within which it can grow wild, otherwise it would have escaped much earlier. It occurs in old fields and along fences within agricultural landscapes, but has been recorded in native woodland, prairie and alvar habitats. It dominates abandoned fields and pasture in some areas and may also increase in certain natural habitats and compete with native species. It is, however, less aggressive than some other alien woody plants and provides abundant food in the form of pollen, nectar and fruit during the early stages of old field succession. Spread is believed to be mostly by humans, but Racoons, deer, and other small mammals may be important in local dispersal

INVESTIGATIONS OF NEW ENGLAND MARINE ALGAE VI: DISTRIBUTION OF MARINE ALGAE NEAR CAPE COD, MASSACHUSETTS

Volume: 76Start Page: 537End Page: 56

Ecoulement dans une géométrie plan-plan confinée: validité de la géométrie pour l'étude de fluides viscoélastiques et application à la dispersion du noir de carbone

Congrès Annuel de la société Européenne de Rhéologie (AERC 2010), Göteborg , Suède (07/04/2010)International audienceSi avoir un écoulement stable est une condition nécessaire pour faire des mesures de rhéologie en régime continu, il en est de même lors d'études rhéo-optiques où des observations d'évolution de structure ou de comportement de particules ou inclusions en suspension dans une matrice sont réalisées in-situ pendant le cisaillement. Dans cet article, nous montrons comment l'installation d'une bague de confinement autour d'une cellule de cisaillement plan-plan contra-rotative transparente permet de réduire le développement d'instabilités d'écoulement dans une matrice polymère viscoélastique pendant le cisaillement, sans modifier l'écoulement sur une zone relativement large de la cellule. Des mesures de périodes de rotation de billes de verre et de rhéologie en régime continu ont été effectuées pour montrer la validité de cette géométrie d'écoulement. Cette cellule a été utilisée pour déterminer les conditions de dispersion d'une charge de noir de carbone dans une matrice polymère sous l'action d'un champ de cisaillement. La détermination des conditions de rupture d'un noir de carbone en suspension dans une matrice polymère dans deux géométries d'écoulement différentes (avec et sans bague de confinement) permet de montrer que le critère de rupture s'exprime bien en terme de contrainte hydrodynamique. L'utilisation de la bague a permis d'atteindre des taux de cisaillement plus élevés dans des conditions stables d'écoulement

Role of AMP-activated protein kinase in adipose tissue metabolism and inflammation

AMPK (AMP-activated protein kinase) is a key regulator of cellular and whole-body energy balance. AMPK phosphorylates and regulates many proteins concerned with nutrient metabolism, largely acting to suppress anabolic ATP-consuming pathways while stimulating catabolic ATP-generating pathways. This has led to considerable interest in AMPK as a therapeutic target for the metabolic dysfunction observed in obesity and insulin resistance. The role of AMPK in skeletal muscle and the liver has been extensively studied, such that AMPK has been demonstrated to inhibit synthesis of fatty acids, cholesterol and isoprenoids, hepatic gluconeogenesis and translation while increasing fatty acid oxidation, muscle glucose transport, mitochondrial biogenesis and caloric intake. The role of AMPK in the other principal metabolic and insulin-sensitive tissue, adipose, remains poorly characterized in comparison, yet increasing evidence supports an important role for AMPK in adipose tissue function. Obesity is characterized by hypertrophy of adipocytes and the development of a chronic sub-clinical pro-inflammatory environment in adipose tissue, leading to increased infiltration of immune cells. This combination of dysfunctional hypertrophic adipocytes and a pro-inflammatory environment contributes to insulin resistance and the development of Type 2 diabetes. Exciting recent studies indicate that AMPK may not only influence metabolism in adipocytes, but also act to suppress this pro-inflammatory environment, such that targeting AMPK in adipose tissue may be desirable to normalize adipose dysfunction and inflammation. In the present review, we discuss the role of AMPK in adipose tissue, focussing on the regulation of carbohydrate and lipid metabolism, adipogenesis and pro-inflammatory pathways in physiological and pathophysiological conditions

Notes on Succession in Old Fields in Southeastern Ontario: the Herbs

Vegetation in abandoned hayfields was monitored during 1976-1998. An earlier successional stage followed ploughing. Changes in tree, shrub and vine populations have been reported earlier and showed expected increases in species richness and cover. Highest species richness of herbs occurred three years after ploughing. Non-woody species richness trended irregularly downward, while non-woody cover was variable, peaking in 1987. Within the herbaceous community, year-to-year changes in cover and frequency of species in the following selected groups are reported here: 18 grasses including sown and adventive species; 13 legumes including two sown species; 14 macroforbs of the Compositae, including a goldenrod, Solidago canadensis, which dominated parts of the fields; a rosette weed, Taraxacum officinalis; sedges, horsetails and some other minor components. Grasses and goldenrods were grazed, sometimes intensively and repeatedly, by insects; grasses were impacted by skipper larvae (Thymelicus lineola), and goldenrods by beetle larvae (Trirhabda spp.). Effects of repeated outbreaks on host plant cover are shown for two plots (100 m2) matching the scale of outbreaks

Fipronil Promotes Adipogenesis via AMPKα-mediated Pathway in 3T3-L1 Adipocytes

Emerging evidence suggests that organochlorine, organophosphorus and neonicotinoid insecticide exposure may be linked to the development of obesity and type 2 diabetes. However, there is no knowledge of the potential influence of fipronil, which belongs to the phenylpyrazole chemical family, on obesity. Thus, the goal of this study was to determine the role of fipronil in adipogenesis using 3T3-L1 adipocytes. Fipronil treatment, at 10 mM, increased fat accumulation in 3T3-L1 adipocytes as well as promoted key regulators of adipocyte differentiation (CCAAT/enhancer-binding protein α and peroxisome proliferator-activated receptor gamma-γ), and key regulators of lipogenesis (acetyl-CoA carboxylase and fatty acid synthase). The activation of AMPKα with 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) abolished effects of fipronil on increased adipogenesis. These results suggest that fipronil alters adipogenesis and results in increased lipid accumulation through a AMPKα-mediated pathway

Characterising the inhibitory actions of ceramide upon insulin signaling in different skeletal muscle cell models:a mechanistic insight

International audienceCeramides are known to promote insulin resistance in a number of metabolically important tissues including skeletal muscle, the predominant site of insulin-stimulated glucose disposal. Depending on cell type, these lipid intermediates have been shown to inhibit protein kinase B (PKB/Akt), a key mediator of the metabolic actions of insulin, via two distinct pathways: one involving the action of atypical protein kinase C (aPKC) isoforms, and the second dependent on protein phosphatase-2A (PP2A). The main aim of this study was to explore the mechanisms by which ceramide inhibits PKB/Akt in three different skeletal muscle-derived cell culture models; rat L6 myotubes, mouse C2C12 myotubes and primary human skeletal muscle cells. Our findings indicate that the mechanism by which ceramide acts to repress PKB/Akt is related to the myocellular abundance of caveolin-enriched domains (CEM) present at the plasma membrane. Here, we show that ceramide-enriched-CEMs are markedly more abundant in L6 myotubes compared to C2C12 myotubes, consistent with their previously reported role in coordinating aPKC-directed repression of PKB/Akt in L6 muscle cells. In contrast, a PP2A-dependent pathway predominantly mediates ceramide-induced inhibition of PKB/Akt in C2C12 myotubes. In addition, we demonstrate for the first time that ceramide engages an aPKC-dependent pathway to suppress insulin-induced PKB/Akt activation in palmitate-treated cultured human muscle cells as well as in muscle cells from diabetic patients. Collectively, this work identifies key mechanistic differences, which may be linked to variations in plasma membrane composition, underlying the insulin-desensitising effects of ceramide in different skeletal muscle cell models that are extensively used in signal transduction and metabolic studies