Store-operated calcium entry and increased endothelial cell permeability

2000 ◽  
Vol 279 (5) ◽  
pp. L815-L824 ◽  
Author(s):  
Natalie Norwood ◽  
Timothy M. Moore ◽  
David A. Dean ◽  
Rakesh Bhattacharjee ◽  
Ming Li ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Calcium Current ◽  
Calcium Release ◽  
Calcium Entry ◽  
Extracellular Calcium ◽  
Cell Permeability ◽  
Calcium Stores ◽  
Store Depletion ◽  
Endothelial Cell Permeability ◽  
Store Operated Calcium Entry

We hypothesized that myosin light chain kinase (MLCK) links calcium release to activation of store-operated calcium entry, which is important for control of the endothelial cell barrier. Acute inhibition of MLCK caused calcium release from inositol trisphosphate-sensitive calcium stores and prevented subsequent activation of store-operated calcium entry by thapsigargin, suggesting that MLCK serves as an important mechanism linking store depletion to activation of membrane calcium channels. Moreover, in voltage-clamped single rat pulmonary artery endothelial cells, thapsigargin activated an inward calcium current that was abolished by MLCK inhibition. F-actin disruption activated a calcium current, and F-actin stabilization eliminated the thapsigargin-induced current. Thapsigargin increased endothelial cell permeability in the presence, but not in the absence, of extracellular calcium, indicating the importance of calcium entry in decreasing barrier function. Although MLCK inhibition prevented thapsigargin from stimulating calcium entry, it did not prevent thapsigargin from increasing permeability. Rather, inhibition of MLCK activity increased permeability that was especially prominent in low extracellular calcium. In conclusion, MLCK links store depletion to activation of a store-operated calcium entry channel. However, inhibition of calcium entry by MLCK is not sufficient to prevent thapsigargin from increasing endothelial cell permeability.

scholarly journals Relocalization of STIM1 in mouse oocytes at fertilization: early involvement of store-operated calcium entry

Reproduction ◽  
2009 ◽  
Vol 138 (2) ◽  
pp. 211-221 ◽  
Author(s):  
Carolina Gómez-Fernández ◽  
Eulalia Pozo-Guisado ◽  
Miguel Gañán-Parra ◽  
Mario J Perianes ◽  
Ignacio S Álvarez ◽  
...  
Keyword(s):  
Intracellular Signaling ◽  
Calcium Entry ◽  
Calcium Stores ◽  
Calcium Waves ◽  
Resting Cells ◽  
Metaphase Arrest ◽  
Mouse Oocytes ◽  
Store Depletion ◽  
Store Operated Calcium Entry ◽  
Mature Mouse

Calcium waves represent one of the most important intracellular signaling events in oocytes at fertilization required for the exit from metaphase arrest and the resumption of the cell cycle. The molecular mechanism ruling this signaling has been described in terms of the contribution of intracellular calcium stores to calcium spikes. In this work, we considered the possible contribution of store-operated calcium entry (SOCE) to this signaling, by studying the localization of the protein STIM1 in oocytes. STIM1 has been suggested to play a key role in the recruitment and activation of plasma membrane calcium channels, and we show here that mature mouse oocytes express this protein distributed in discrete clusters throughout their periphery in resting cells, colocalizing with the endoplasmic reticulum marker calreticulin. However, immunolocalization of the endogenous STIM1 showed considerable redistribution over larger areas or patches covering the entire periphery of the oocyte during Ca2+ store depletion induced with thapsigargin or ionomycin. Furthermore, pharmacological activation of endogenous phospholipase C induced a similar pattern of redistribution of STIM1 in the oocyte. Finally, fertilization of mouse oocytes revealed a significant and rapid relocalization of STIM1, similar to that found after pharmacological Ca2+ store depletion. This particular relocalization supports a role for STIM1 and SOCE in the calcium signaling during early stages of fertilization.

scholarly journals An essential and NSF independent role for α-SNAP in store-operated calcium entry

eLife ◽  
2013 ◽  
Vol 2 ◽  
Author(s):  
Yong Miao ◽  
Cathrine Miner ◽  
Lei Zhang ◽  
Phyllis I Hanson ◽  
Adish Dani ◽  
...  
Keyword(s):  
Essential Role ◽  
Calcium Release ◽  
Calcium Entry ◽  
Snare Complex ◽  
Functional Coupling ◽  
Crac Channels ◽  
Store Depletion ◽  
Store Operated Calcium Entry ◽  
Crac Channel

Store-operated calcium entry (SOCE) by calcium release activated calcium (CRAC) channels constitutes a primary route of calcium entry in most cells. Orai1 forms the pore subunit of CRAC channels and Stim1 is the endoplasmic reticulum (ER) resident Ca2+ sensor. Upon store-depletion, Stim1 translocates to domains of ER adjacent to the plasma membrane where it interacts with and clusters Orai1 hexamers to form the CRAC channel complex. Molecular steps enabling activation of SOCE via CRAC channel clusters remain incompletely defined. Here we identify an essential role of α-SNAP in mediating functional coupling of Stim1 and Orai1 molecules to activate SOCE. This role for α-SNAP is direct and independent of its known activity in NSF dependent SNARE complex disassembly. Importantly, Stim1-Orai1 clustering still occurs in the absence of α-SNAP but its inability to support SOCE reveals that a previously unsuspected molecular re-arrangement within CRAC channel clusters is necessary for SOCE.

scholarly journals Receptor-dependent activation of store-operated calcium entry increases endothelial cell permeability

2000 ◽  
Vol 279 (4) ◽  
pp. L691-L698 ◽  
Author(s):  
Timothy M. Moore ◽  
Natalie R. Norwood ◽  
Judy R. Creighton ◽  
Pavál Babal ◽  
George H. Brough ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Myosin Light Chain ◽  
Cell Permeability ◽  
Thrombin Receptor ◽  
Gap Formation ◽  
Common Pathway ◽  
Endothelial Cell Permeability ◽  
Store Operated Calcium Entry ◽  
Transport Of Macromolecules ◽  
Sustained Increase

The present study evaluated the necessity of store-operated Ca2+ entry in mediating thrombin-induced 20-kDa myosin light chain (MLC20) phosphorylation and increased permeability in bovine pulmonary artery endothelial cells (BPAECs). Thrombin (7 U/ml) and thapsigargin (1 μM) activated Ca2+ entry through a common pathway in confluent BPAECs. Similar increases in MLC20 phosphorylation were observed 5 min after thrombin and thapsigargin challenge, although thrombin produced a sustained increase in MLC20phosphorylation that was not observed in response to thapsigargin. Neither agonist increased MLC20 phosphorylation when Ca2+ influx was inhibited. Thrombin and thapsigargin induced inter-endothelial cell gap formation and increased FITC-dextran (molecular radii 23 Å) transfer across confluent BPAEC monolayers. Activation of store-operated Ca2+ entry was required for thapsigargin and thrombin receptor-activating peptide to increase permeability, demonstrating that activation of store-operated Ca2+ entry is coupled with MLC20phosphorylation and is associated with intercellular gap formation and increased barrier transport of macromolecules. Unlike thrombin receptor-activating peptide, thrombin increased permeability without activation of store-operated Ca2+ entry, suggesting that it partly disrupts the endothelial barrier through a proteolytic mechanism independent of Ca2+ signaling.

scholarly journals Junctate is a key element in calcium entry induced by activation of InsP3 receptors and/or calcium store depletion

2004 ◽  
Vol 166 (4) ◽  
pp. 537-548 ◽  
Author(s):  
Susan Treves ◽  
Clara Franzini-Armstrong ◽  
Luca Moccagatta ◽  
Christophe Arnoult ◽  
Cristiano Grasso ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Intracellular Calcium ◽  
Calcium Binding ◽  
Calcium Release ◽  
Transient Receptor Potential ◽  
Receptor Activation ◽  
Calcium Entry ◽  
Calcium Stores ◽  
Store Depletion ◽  
Insp3 Receptors

In many cell types agonist-receptor activation leads to a rapid and transient release of Ca2+ from intracellular stores via activation of inositol 1,4,5 trisphosphate (InsP3) receptors (InsP3Rs). Stimulated cells activate store- or receptor-operated calcium channels localized in the plasma membrane, allowing entry of extracellular calcium into the cytoplasm, and thus replenishment of intracellular calcium stores. Calcium entry must be finely regulated in order to prevent an excessive intracellular calcium increase. Junctate, an integral calcium binding protein of endo(sarco)plasmic reticulum membrane, (a) induces and/or stabilizes peripheral couplings between the ER and the plasma membrane, and (b) forms a supramolecular complex with the InsP3R and the canonical transient receptor potential protein (TRPC) 3 calcium entry channel. The full-length protein modulates both agonist-induced and store depletion–induced calcium entry, whereas its NH2 terminus affects receptor-activated calcium entry. RNA interference to deplete cells of endogenous junctate, knocked down both agonist-activated calcium release from intracellular stores and calcium entry via TRPC3. These results demonstrate that junctate is a new protein involved in calcium homeostasis in eukaryotic cells.

Signal transduction and regulation of lung endothelial cell permeability. Interaction between calcium and cAMP

1998 ◽  
Vol 275 (2) ◽  
pp. L203-L222 ◽  
Author(s):  
Timothy M. Moore ◽  
Paul M. Chetham ◽  
John J. Kelly ◽  
Troy Stevens
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Intracellular Signaling ◽  
Endothelial Permeability ◽  
Cell Permeability ◽  
Gap Formation ◽  
Pulmonary Endothelium ◽  
Intracellular Signals ◽  
Endothelial Cell Permeability ◽  
Cell Cell

Pulmonary endothelium forms a semiselective barrier that regulates fluid balance and leukocyte trafficking. During the course of lung inflammation, neurohumoral mediators and oxidants act on endothelial cells to induce intercellular gaps permissive for transudation of proteinaceous fluid from blood into the interstitium. Intracellular signals activated by neurohumoral mediators and oxidants that evoke intercellular gap formation are incompletely understood. Cytosolic Ca2+ concentration ([Ca2+]i) and cAMP are two signals that importantly dictate cell-cell apposition. Although increased [Ca2+]ipromotes disruption of the macrovascular endothelial cell barrier, increased cAMP enhances endothelial barrier function. Furthermore, during the course of inflammation, elevated endothelial cell [Ca2+]idecreases cAMP to facilitate intercellular gap formation. Given the significance of both [Ca2+]iand cAMP in mediating cell-cell apposition, this review addresses potential sites of cross talk between these two intracellular signaling pathways. Emerging data also indicate that endothelial cells derived from different vascular sites within the pulmonary circulation exhibit distinct sensitivities to permeability-inducing stimuli; that is, elevated [Ca2+]ipromotes macrovascular but not microvascular barrier disruption. Thus this review also considers the roles of [Ca2+]iand cAMP in mediating site-specific alterations in endothelial permeability.

Novel infection of pericytes by Andes virus enhances endothelial cell permeability

2021 ◽  
Vol 306 ◽  
pp. 198584
Author(s):  
Ramon D. Perez ◽  
Elena E. Gorbonova ◽  
Erich R. Mackow
Keyword(s):  
Endothelial Cell ◽  
Cell Permeability ◽  
Andes Virus ◽  
Endothelial Cell Permeability

scholarly journals Serine/threonine phosphatase 5 (PP5C/PPP5C) regulates the ISOC channel through a PP5C-FKBP51 axis

2017 ◽  
Vol 8 (1) ◽  
pp. 204589321775315 ◽  
Author(s):  
Caleb L. Hamilton ◽  
Kevin A. Abney ◽  
Audrey A. Vasauskas ◽  
Mikhail Alexeyev ◽  
Li Ni ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Heat Shock Protein 90 ◽  
Calcium Entry ◽  
Hek293 Cells ◽  
Wide Field ◽  
Cell Fractionation ◽  
Gap Formation ◽  
Tetratricopeptide Repeats ◽  
Store Operated Calcium Entry

Pulmonary endothelial cells express a store-operated calcium entry current ( Isoc), which contributes to inter-endothelial cell gap formation. Isoc is regulated by a heterocomplex of proteins that includes the immunophilin FKBP51. FKBP51 inhibits Isoc by mechanisms that are not fully understood. In pulmonary artery endothelial cells (PAECs) we have shown that FKBP51 increases microtubule polymerization, an event that is critical for Isoc inhibition by FKBP51. In neurons, FKBP51 promotes microtubule stability through facilitation of tau dephosphorylation. However, FKBP51 does not possess phosphatase activity. Protein phosphatase 5 (PP5C/PPP5C) can dephosphorylate tau, and similar to FKBP51, PP5C possesses tetratricopeptide repeats (TPR) that mediate interaction with heat shock protein-90 (HSP90) chaperone/scaffolding complexes. We therefore tested whether PP5C contributes to FKBP51-mediated inhibition of Isoc. Both siRNA-mediated suppression of PP5C expression in PAECs and genetic disruption of PP5C in HEK293 cells attenuate FKBP51-mediated inhibition of Isoc. Reintroduction of catalytically competent, but not catalytically inactive PP5C, restored FKBP51-mediated inhibition of Isoc. PAEC cell fractionation studies identified both PP5C and the ISOC heterocomplex in the same membrane fractions. Further, PP5C co-precipitates with TRPC4, an essential subunit of ISOC channel. Finally, to determine if PP5C is required for FKBP51-mediated inhibition of calcium entry-induced inter-endothelial cell gap formation, we measured gap area by wide-field microscopy and performed biotin gap quantification assay and electric cell-substrate impedance sensing (ECIS®). Collectively, the data presented indicate that suppression of PP5C expression negates the protective effect of FKBP51. These observations identify PP5C as a novel member of the ISOC heterocomplex that is required for FKBP51-mediated inhibition of Isoc.

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