Signaling pathways in cytoskeletal responses to plasma membrane depolarization in corneal endothelial cells

2019 ◽  
Vol 235 (3) ◽  
pp. 2947-2962
Author(s):  
Frances Evans ◽  
Julio A. Hernández ◽  
Silvia Chifflet
Keyword(s):  
Endothelial Cells ◽  
Plasma Membrane ◽  
Signaling Pathways ◽  
Membrane Depolarization ◽  
Corneal Endothelial Cells ◽  
Cytoskeletal Responses

Oxidative burst and NO generation as initial response to ischemia in flow-adapted endothelial cells

2001 ◽  
Vol 280 (5) ◽  
pp. H2126-H2135 ◽  
Author(s):  
Yefim Manevich ◽  
Abu Al-Mehdi ◽  
Vladimir Muzykantov ◽  
Aron B. Fisher
Keyword(s):  
Endothelial Cells ◽  
Plasma Membrane ◽  
Oxidative Burst ◽  
Initial Response ◽  
Membrane Depolarization ◽  
No Synthase ◽  
Potential Production ◽  
Perfusion Chamber ◽  
Intracellular Ca ◽  
Static Conditions

Shear stress modulates endothelial physiology, yet the effect(s) of flow cessation is poorly understood. The initial metabolic responses of flow-adapted bovine pulmonary artery endothelial cells to the abrupt cessation of flow (simulated ischemia) was evaluated using a perfusion chamber designed for continuous spectroscopy. Plasma membrane potential, production of reactive O2 species (ROS), and intracellular Ca2+ and nitric oxide (NO) levels were measured with fluorescent probes. Within 15 s after flow cessation, flow-adapted cells, but not cells cultured under static conditions, showed plasma membrane depolarization and an oxidative burst with generation of ROS that was inhibited by diphenyleneiodonium. EGTA-inhibitable elevation of intracellular Ca2+ and NO were observed at ∼30 and 60 s after flow cessation, respectively. NO generation was decreased in the presence of inhibitors of NO synthase and calmodulin. Thus flow-adapted endothelial cells sense the altered hemodynamics associated with flow cessation and respond by plasma membrane depolarization, activation of NADPH oxidase, Ca2+ influx, and activation of Ca2+/calmodulin-dependent NO synthase. This signaling response is unrelated to cellular anoxia.

Membrane depolarization and NADPH oxidase activation in aortic endothelium during ischemia reflect altered mechanotransduction

2005 ◽  
Vol 288 (1) ◽  
pp. H336-H343 ◽  
Author(s):  
Ikuo Matsuzaki ◽  
Shampa Chatterjee ◽  
Kris DeBolt ◽  
Yefim Manevich ◽  
Qunwei Zhang ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Plasma Membrane ◽  
Membrane Potential ◽  
Nadph Oxidase ◽  
Membrane Depolarization ◽  
Ros Generation ◽  
Similar Response ◽  
Aortic Endothelial Cells ◽  
Aortic Endothelium ◽  
Aortic Endothelial

We previously showed that “ischemia” (abrupt cessation of flow) leads to rapid membrane depolarization and increased generation of reactive oxygen species (ROS) in lung microvascular endothelial cells. This response is not associated with anoxia but, rather, reflects loss of normal shear stress. This study evaluated whether a similar response occurs in aortic endothelium. Plasma membrane potential and production of ROS were determined by fluorescence microscopy and cytochrome c reduction in flow-adapted rat or mouse aorta or monolayer cultures of rat aortic endothelial cells. Within 30 s after flow cessation, endothelial cells that had been flow adapted showed plasma membrane depolarization that was inhibited by pretreatment with cromakalim, an ATP-sensitive K+ (KATP) channel agonist. Flow cessation also led to ROS generation, which was inhibited by cromakalim and the flavoprotein inhibitor diphenyleneiodonium. Aortic endothelium from mice with “knockout” of the KATP channel (KIR6.2) showed a markedly attenuated change in membrane potential and ROS generation with flow cessation. In aortic endothelium from mice with knockout of NADPH oxidase (gp91phox), membrane depolarization was similar to that in wild-type mice but ROS generation was absent. Thus rat and mouse aortic endothelial cells respond to abrupt flow cessation by KATP channel-mediated membrane depolarization followed by NADPH oxidase-mediated ROS generation, possibly representing a cell-signaling response to altered mechanotransduction.

Signaling pathways mediating VEGF 165 ‐induced calcium transients and membrane depolarization in human endothelial cells

2006 ◽  
Vol 20 (7) ◽  
pp. 991-993 ◽  
Author(s):  
Nancy S. Dawson ◽  
David C. Zawieja ◽  
Mack H. Wu ◽  
Harris J. Granger ◽  
Nancy S. Dawson ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Signaling Pathways ◽  
Membrane Depolarization ◽  
Calcium Transients ◽  
Human Endothelial Cells

Culturing Discarded Peripheral Human Corneal Endothelial Cells From the Tissues Deemed for Preloaded DMEK Transplants

Cornea ◽  
2019 ◽  
Vol 38 (9) ◽  
pp. 1175-1181 ◽  
Author(s):  
Mohit Parekh ◽  
Vito Romano ◽  
Alessandro Ruzza ◽  
Stephen B. Kaye ◽  
Diego Ponzin ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Corneal Endothelial Cells ◽  
Human Corneal Endothelial Cells
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