Clathrin-dependent entry and vesicle-mediated exocytosis define insulin transcytosis across microvascular endothelial cells

Azizi, Paymon; Zyla, Roman; Guan, Sha; Wang, Changsen; Li, Jun; Bolz, Steffen‐Sebastian; Heit, Bryan; Klip, Amira; Lee, Warren L.

doi:10.1091/mbc.e14-08-1307

Cited by 75 publications

(85 citation statements)

References 40 publications

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“…The phenotypic contrast between microvascular and macrovascular ECs (34) creates difficulties in interpretation. In fact, the mechanism by which insulin is taken up by ECs depends on the type of EC used (35). who previously showed that the delivery of insulin to SkM is not saturable in dogs (12).…”

Section: Discussionmentioning

confidence: 99%

Insulin exits skeletal muscle capillaries by fluid-phase transport

Williams¹,

Valenzuela²,

Kahl³

et al. 2018

Journal of Clinical Investigation

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Section: Discussionmentioning

confidence: 99%

Insulin exits skeletal muscle capillaries by fluid-phase transport

Williams¹,

Valenzuela²,

Kahl³

et al. 2018

Journal of Clinical Investigation

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“…For the staining of viral nucleoprotein and PECAM-1, antigen retrieval was performed by boiling for 15 min in citric acid buffer, cooling for 45 min, and then blocking and permeabilization. Lung sections were stained with rabbit anti-PECAM-1 (1:500) overnight and with mouse anti-influenza NP (1:75) for 1 h. The colocalization of viral NP and PECAM-1 was quantified using Manders' coefficient (27) as determined using Volocity software.…”

Section: Methodsmentioning

confidence: 99%

Influenza Virus Infection Induces Platelet-Endothelial Adhesion Which Contributes to Lung Injury

Sugiyama

Gamage

Zyla

et al. 2016

J Virol

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Lung injury after influenza infection is characterized by increased permeability of the lung microvasculature, culminating in acute respiratory failure. Platelets interact with activated endothelial cells and have been implicated in the pathogenesis of some forms of acute lung injury. Autopsy studies have revealed pulmonary microthrombi after influenza infection, and epidemiological studies suggest that influenza vaccination is protective against pulmonary thromboembolism; however, the effect of influenza infection on platelet-endothelial interactions is unclear. We demonstrate that endothelial infection with both laboratory and clinical strains of influenza virus increased the adhesion of human platelets to primary human lung microvascular endothelial cells. Platelets adhered to infected cells as well as to neighboring cells, suggesting a paracrine effect. Influenza infection caused the upregulation of von Willebrand factor and ICAM-1, but blocking these receptors did not prevent platelet-endothelial adhesion. Instead, platelet adhesion was inhibited by both RGDS peptide and a blocking antibody to platelet integrin ␣ 5 ␤ 1 , implicating endothelial fibronectin. Concordantly, lung histology from infected mice revealed viral dose-dependent colocalization of viral nucleoprotein and the endothelial marker PECAM-1, while platelet adhesion and fibronectin deposition also were observed in the lungs of influenza-infected mice. Inhibition of platelets using acetylsalicylic acid significantly improved survival, a finding confirmed using a second antiplatelet agent. Thus, influenza infection induces platelet-lung endothelial adhesion via fibronectin, contributing to mortality from acute lung injury. The inhibition of platelets may constitute a practical adjunctive strategy to the treatment of severe infections with influenza. Here, we show that this infection causes platelets to adhere to the lung endothelium. Importantly, blocking platelets using two distinct antiplatelet drugs improved survival in a mouse model of severe influenza infection. Thus, platelet inhibition may constitute a novel therapeutic strategy to improve the host response to severe infections with influenza.

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“…Transcytosis of albumin was detected using total internal reflection fluorescence microscopy, as recently described (28). Briefly, rhodamine-albumin (Sigma-Aldrich) was allowed to bind to the apical membrane at 4°C for 10 min, after which excess albumin was washed off.…”

Section: Albumin Transcytosis Assaymentioning

confidence: 99%

Prostaglandin I2 Receptor Agonism Preserves β-Cell Function and Attenuates Albuminuria Through Nephrin-Dependent Mechanisms

et al. 2016

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Discovery of common pathways that mediate both pancreatic b-cell function and end-organ function offers the opportunity to develop therapies that modulate glucose homeostasis and separately slow the development of diabetes complications. Here, we investigated the in vitro and in vivo effects of pharmacological agonism of the prostaglandin I 2 (IP) receptor in pancreatic b-cells and in glomerular podocytes. The IP receptor agonist MRE-269 increased intracellular 39,59-cyclic adenosine monophosphate (cAMP), augmented glucose-stimulated insulin secretion (GSIS), and increased viability in MIN6 b-cells. Its prodrug form, selexipag, augmented GSIS and preserved islet b-cell mass in diabetic mice. Determining that this preservation of b-cell function is mediated through cAMP/protein kinase A (PKA)/ nephrin-dependent pathways, we found that PKA inhibition, nephrin knockdown, or targeted mutation of phosphorylated nephrin tyrosine residues 1176 and 1193 abrogated the actions of MRE-269 in MIN6 cells. Because nephrin is important to glomerular permselectivity, we next set out to determine whether IP receptor agonism similarly affects nephrin phosphorylation in podocytes. Expression of the IP receptor in podocytes was confirmed in cultured cells by immunoblotting and quantitative real-time PCR and in mouse kidneys by immunogold electron microscopy, and its agonism 1) increased cAMP, 2) activated PKA, 3) phosphorylated nephrin, and 4) attenuated albumin transcytosis. Finally, treatment of diabetic endothelial nitric oxide synthase knockout mice with selexipag augmented renal nephrin phosphorylation and attenuated albuminuria development independently of glucose change. Collectively, these observations describe a pharmacological strategy that posttranslationally modifies nephrin and the effects of this strategy in the pancreas and in the kidney.

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Clathrin-dependent entry and vesicle-mediated exocytosis define insulin transcytosis across microvascular endothelial cells

Cited by 75 publications

References 40 publications

Insulin exits skeletal muscle capillaries by fluid-phase transport

Insulin exits skeletal muscle capillaries by fluid-phase transport

Influenza Virus Infection Induces Platelet-Endothelial Adhesion Which Contributes to Lung Injury

Prostaglandin I2 Receptor Agonism Preserves β-Cell Function and Attenuates Albuminuria Through Nephrin-Dependent Mechanisms

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