Lack of CD2AP disrupts Glut4 trafficking and attenuates glucose uptake in podocytes

Tolvanen, Tuomas A.; Dash, S. K.; Polianskyte-Prause, Zydrune; Dumont, Vincent; Lehtonen, Sanna

doi:10.1242/jcs.175075

Cited by 18 publications

(26 citation statements)

References 66 publications

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“…L6 myoblasts stably expressing hemagglutinin-GLUT4-green fluorescent protein (HA-GLUT4-GFP, referred to as L6-GLUT4) were generated, as previously described (19). Immortalized human podocytes were maintained as previously described (10).…”

Section: Methodsmentioning

confidence: 99%

“…Rat L6 myoblasts (American Type Culture Collection, Manassas, VA, USA) and hepatoma cells (Fao Cells, Culture Collection; Public Health England, Salisbury, United Kingdom) were maintained according to the provider's instructions. L6 myoblasts stably expressing hemagglutinin-GLUT4-green fluorescent protein (HA-GLUT4-GFP, referred to as L6-GLUT4) were generated, as previously described (19). Immortalized human podocytes were maintained as previously described (10).…”

Section: Cell Culture and Biochemical Assaysmentioning

confidence: 99%

“…Podocytes were lysed as previously described (21). Myotubes, hepatoma cells, and tissues were lysed and immunoblotting performed as in Tolvanen et al (19) with anti-SHIP2 and anti-AMPK (Santa Cruz Biotechnology, Dallas, TX, USA); anti-phospho (p)Akt (S473), anti-pAMPK (Thr172), anti-AMPK, and anti-cleaved caspase-3 (Cell Signaling Technology, Danvers, MA, USA); anti-Pan Akt (R&D Systems, Minneapolis, MN, USA); anti-tubulin, anti-Glut4, and anti-Glut1 (Millipore-Sigma, Burlington, MA, USA); and anti-actin (Abcam, Cambridge, United Kingdom). Immunofluorescence and cell-surface labeling with anti-HA.11 (Covance, Princeton, NJ, USA) were performed as in Tolvanen et al (19) and Heikkila et al (21) using CAS-Block (Vector Laboratories, Burlingame, CA, USA).…”

Section: Cell Culture and Biochemical Assaysmentioning

confidence: 99%

“…Glucose uptake assay and On-Cell Western were performed essentially as in Tolvanen et al (19). For HA-GLUT4-GFP endocytosis assay, L6-GLUT4 cells were incubated under basal conditions with metformin (2 mM, 20-24 h) or stimulated with insulin (100 nM, 15 min).…”

Section: Glucose Uptake and Glut4 Translocation Assaysmentioning

confidence: 99%

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Metformin increases glucose uptake and acts renoprotectively by reducing SHIP2 activity

Polianskyte-Prause¹,

Tolvanen²,

Lindfors³

et al. 2018

FASEB j.

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Metformin, the first-line drug to treat type 2 diabetes (T2D), inhibits mitochondrial glycerolphosphate dehydrogenase in the liver to suppress gluconeogenesis. However, the direct target and the underlying mechanisms by which metformin increases glucose uptake in peripheral tissues remain uncharacterized. Lipid phosphatase Src homology 2 domain-containing inositol-5-phosphatase 2 (SHIP2) is upregulated in diabetic rodent models and suppresses insulin signaling by reducing Akt activation, leading to insulin resistance and diminished glucose uptake. Here, we demonstrate that metformin directly binds to and reduces the catalytic activity of the recombinant SHIP2 phosphatase domain in vitro. Metformin inhibits SHIP2 in cultured cells and in skeletal muscle and kidney of db/db mice. In SHIP2-overexpressing myotubes, metformin ameliorates reduced glucose uptake by slowing down glucose transporter 4 endocytosis. SHIP2 overexpression reduces Akt activity and enhances podocyte apoptosis, and both are restored to normal levels by metformin. SHIP2 activity is elevated in glomeruli of patients with T2D receiving nonmetformin medication, but not in patients receiving metformin, compared with people without diabetes. Furthermore, podocyte loss in kidneys of metformin-treated T2D patients is reduced compared with patients receiving nonmetformin medication. Our data unravel a novel molecular mechanism by which metformin enhances glucose uptake and acts renoprotectively by reducing SHIP2 activity.—Polianskyte-Prause, Z., Tolvanen, T. A., Lindfors, S., Dumont, V., Van, M., Wang, H., Dash, S. N., Berg, M., Naams, J.-B., Hautala, L. C., Nisen, H., Mirtti, T., Groop, P.-H., Wähälä, K., Tienari, J., Lehtonen, S. Metformin increases glucose uptake and acts renoprotectively by reducing SHIP2 activity.

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Section: Methodsmentioning

confidence: 99%

Section: Cell Culture and Biochemical Assaysmentioning

confidence: 99%

Section: Cell Culture and Biochemical Assaysmentioning

confidence: 99%

Section: Glucose Uptake and Glut4 Translocation Assaysmentioning

confidence: 99%

See 2 more Smart Citations

Metformin increases glucose uptake and acts renoprotectively by reducing SHIP2 activity

Polianskyte-Prause¹,

Tolvanen²,

Lindfors³

et al. 2018

FASEB j.

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“…We observed that endocytosed nephrin was not trapped in the endolysosomal system (Supplemental Fig. 2), but the punctate structures stained with antibodies against nephrin were also positive for CD2AP, which regulates intracellular trafficking in podocytes (35) and anchors the slit diaphragm to the cytoskeleton by binding actin directly (36). Strikingly, podocin, which escorts newly synthesized nephrin to the plasma membrane (5), also showed extensive colocalization with nephrin on the punctate structures (Supplemental Fig.…”

Section: Discussionmentioning

confidence: 99%

PACSIN2 accelerates nephrin trafficking and is up‐regulated in diabetic kidney disease

et al. 2017

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Nephrin is a core component of podocyte (glomerular epithelial cell) slit diaphragm and is required for kidney ultrafiltration. Down-regulation or mislocalization of nephrin has been observed in diabetic kidney disease (DKD), characterized by albuminuria. Here, we investigate the role of protein kinase C and casein kinase 2 substrate in neurons 2 (PACSIN2), a regulator of endocytosis and recycling, in the trafficking of nephrin and development of DKD. We observe that PACSIN2 is up-regulated and nephrin mislocalized in podocytes of obese Zucker diabetic fatty (ZDF) rats that have altered renal function. In cultured podocytes, PACSIN2 and nephrin colocalize and interact. We show that nephrin is endocytosed in PACSIN2-positive membrane regions and that PACSIN2 overexpression increases both nephrin endocytosis and recycling. We identify rabenosyn-5, which is involved in early endosome maturation and endosomal sorting, as a novel interaction partner of PACSIN2. Interestingly, rabenosyn-5 expression is increased in podocytes in obese ZDF rats, and, in vitro, its overexpression enhances the association of PACSIN2 and nephrin. We also show that palmitate, which is elevated in diabetes, enhances this association. Collectively, PACSIN2 is up-regulated and nephrin is abnormally localized in podocytes of diabetic ZDF rats. In vitro, PACSIN2 enhances nephrin turnover apparently via a mechanism involving rabenosyn-5. The data suggest that elevated PACSIN2 expression accelerates nephrin trafficking and associates with albuminuria

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Dehydration Accelerates Cytogenesis and Cyst Growth in Pkd1−/− Mice by Regulating Macrophage M2 Polarization

et al. 2023

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Adult autosomal dominant polycystic kidney disease (ADPKD) has been shown to be related as a "third hit" to the occurrence of acute or chronic kidney injury. Here, we examined whether dehydration, as a common kidney risk factor, could cause cystogenesis in chronic-onset Pkd1 −/− mice by regulating macrophage activation. First, we con rmed that dehydration accelerated cytogenesis in Pkd1 −/− mice and that macrophages in ltrated the kidney tissues even earlier than macroscopic cyst formation. Then, microarray analysis suggested that glycolysis pathway may be involved in macrophage activation in Pkd1 −/− kidneys under conditions of dehydration. Further, we con rmed glycolysis pathway was activated and lactic acid (L-LA) was overproduced in the Pkd1 −/− kidney under conditions of dehydration. We have already proved that L-LA strongly stimulated M2 macrophage polarization and overproduction of polyamine in macrophage in vitro; and in the present study, we further discovered that M2 polarizationinduced polyamine production shortened the primary cilia length by disrupting the PC1/PC2 complex. Finally, the activation of L-LA-arginase 1-polyamine pathway contributed to cystogenesis and progressive cyst growth in Pkd1 −/− mice recurrently exposed to dehydration.

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Lack of CD2AP disrupts Glut4 trafficking and attenuates glucose uptake in podocytes

Cited by 18 publications

References 66 publications

Metformin increases glucose uptake and acts renoprotectively by reducing SHIP2 activity

Metformin increases glucose uptake and acts renoprotectively by reducing SHIP2 activity

PACSIN2 accelerates nephrin trafficking and is up‐regulated in diabetic kidney disease

Dehydration Accelerates Cytogenesis and Cyst Growth in Pkd1−/− Mice by Regulating Macrophage M2 Polarization

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