Effect of the myosin light chain kinase inhibitor ML-7 on the proteome of hearts subjected to ischemia–reperfusion injury

2012 ◽  
Vol 75 (17) ◽  
pp. 5386-5395 ◽  
Author(s):  
Han-bin Lin ◽  
Virgilio J.J. Cadete ◽  
Jolanta Sawicka ◽  
Mieczyslaw Wozniak ◽  
Grzegorz Sawicki
Keyword(s):  
Reperfusion Injury ◽  
Light Chain ◽  
Myosin Light Chain Kinase ◽  
Myosin Light Chain ◽  
Kinase Inhibitor ◽  
Ischemia Reperfusion Injury ◽  
Ischemia Reperfusion

scholarly journals Degradation of Myosin Light Chain in Isolated Rat Hearts Subjected to Ischemia-Reperfusion Injury

Circulation ◽  
2005 ◽  
Vol 112 (4) ◽  
pp. 544-552 ◽  
Author(s):  
Grzegorz Sawicki ◽  
Hernando Leon ◽  
Jolanta Sawicka ◽  
Meltem Sariahmetoglu ◽  
Costas J. Schulze ◽  
...  
Keyword(s):  
Reperfusion Injury ◽  
Light Chain ◽  
Myosin Light Chain ◽  
Ischemia Reperfusion Injury ◽  
Ischemia Reperfusion ◽  
Rat Hearts ◽  
Isolated Rat

scholarly journals Myosin light chain kinase mediates intestinal barrier dysfunction via occludin endocytosis during anoxia/reoxygenation injury

2016 ◽  
Vol 311 (6) ◽  
pp. C996-C1004 ◽  
Author(s):  
Younggeon Jin ◽  
Anthony T. Blikslager
Keyword(s):  
Tight Junction ◽  
Light Chain ◽  
Barrier Function ◽  
Myosin Light Chain Kinase ◽  
Myosin Light Chain ◽  
Ischemia Reperfusion Injury ◽  
Ischemia Reperfusion ◽  
Barrier Dysfunction ◽  
Junction Protein ◽  
Mlc Phosphorylation

Intestinal anoxia/reoxygenation (A/R) injury induces loss of barrier function followed by epithelial repair. Myosin light chain kinase (MLCK) has been shown to alter barrier function via regulation of interepithelial tight junctions, but has not been studied in intestinal A/R injury. We hypothesized that A/R injury would disrupt tight junction barrier function via MLCK activation and myosin light chain (MLC) phosphorylation. Caco-2BBe1 monolayers were subjected to anoxia for 2 h followed by reoxygenation in 21% O2, after which barrier function was determined by measuring transepithelial electrical resistance (TER) and FITC-dextran flux. Tight junction proteins and MLCK signaling were assessed by Western blotting, real-time PCR, or immunofluorescence microscopy. The role of MLCK was further investigated with select inhibitors (ML-7 and peptide 18) by using in vitro and ex vivo models. Following A/R injury, there was a significant increase in paracellular permeability compared with control cells, as determined by TER and dextran fluxes ( P < 0.05). The tight junction protein occludin was internalized during A/R injury and relocalized to the region of the tight junction after 4 h of recovery. MLC phosphorylation was significantly increased by A/R injury ( P < 0.05), and treatment with the MLCK inhibitor peptide 18 attenuated the increased epithelial monolayer permeability and occludin endocytosis caused by A/R injury. Application of MLCK inhibitors to ischemia-injured porcine ileal mucosa induced significant increases in TER and reduced mucosal-to-serosal fluxes of3H-labeled mannitol. These data suggest that MLCK-induced occludin endocytosis mediates intestinal epithelial barrier dysfunction during A/R injury. Our results also indicate that MLCK-dependent occludin regulation may be a target for the therapeutic treatment of ischemia/reperfusion injury.

L-selectin stimulation of canine neutrophil initiates calcium signal secondary to tyrosine kinase activation

1997 ◽  
Vol 272 (3) ◽  
pp. H1302-H1308 ◽  
Author(s):  
E. Crockett-Torabi ◽  
J. C. Fantone
Keyword(s):  
Tyrosine Kinase ◽  
Reperfusion Injury ◽  
Protein Tyrosine Kinase ◽  
Kinase Inhibitor ◽  
Ischemia Reperfusion Injury ◽  
Leukocyte Adhesion ◽  
Ischemia Reperfusion ◽  
Second Messengers ◽  
Cross Linking ◽  
Calcium Signal

Neutrophils play an important role in myocardial ischemia-reperfusion injury. Neutrophil adhesion to the vascular endothelium is one of the important early mechanisms that lead to reperfusion injury. The leukocyte adhesion molecule, L-selectin, plays a major role in the initial interaction between neutrophils and endothelial cells. Intervention aimed at blocking selectins or their associated ligands can exert cardioprotective effects. The purpose of this study was to examine the role of L-selectin in the initiation of transmembrane signaling and regulation of canine neutrophil responses. Cross-linking of canine neutrophil L-selectin using anti-L-selectin antibody induced a rapid and transient increase in intracellular Ca2+ levels and superoxide anion generation that were dependent on the extent of L-selectin cross-linking. The responses were significantly inhibited by the protein tyrosine kinase inhibitor, genistein. The results demonstrate that ligation of canine neutrophil L-selectin is coupled to intracellular signal transduction pathways and the generation of second messengers, which may independently play important regulatory roles in modulating neutrophil-endothelial cell interactions.

scholarly journals Protective Effect of Inhaled Rho-Kinase Inhibitor on Lung Ischemia-Reperfusion Injury

2017 ◽  
Vol 103 (2) ◽  
pp. 476-483 ◽  
Author(s):  
Keiji Ohata ◽  
Toyofumi F. Chen-Yoshikawa ◽  
Toshi Menju ◽  
Ei Miyamoto ◽  
Satona Tanaka ◽  
...  
Keyword(s):  
Protective Effect ◽  
Reperfusion Injury ◽  
Kinase Inhibitor ◽  
Ischemia Reperfusion Injury ◽  
Ischemia Reperfusion ◽  
Rho Kinase ◽  
Rho Kinase Inhibitor

Role of calmodulin and myosin light-chain kinase in lung ischemia-reperfusion injury

1996 ◽  
Vol 271 (1) ◽  
pp. L121-L125 ◽  
Author(s):  
P. L. Khimenko ◽  
T. M. Moore ◽  
P. S. Wilson ◽  
A. E. Taylor
Keyword(s):  
Endothelial Cell ◽  
Lung Injury ◽  
Light Chain ◽  
Myosin Light Chain Kinase ◽  
Myosin Light Chain ◽  
Ischemia Reperfusion ◽  
Calcium Ionophore ◽  
Microvascular Permeability ◽  
Microvascular Damage ◽  
Pkc Inhibitor

It is generally accepted that microvascular permeability is controlled by intercellular endothelial cell gap size. This process is controlled in endothelial cell monolayers and peripheral blood vessels by calmodulin (CaM)-dependent myosin light-chain kinase (MLCK), which phosphorylates MLC20 with subsequent actin-myosin interaction. In the present study both CaM and MLCK blockers were studied during ischemia-reperfusion (I/R)-induced injury in isolated buffer-perfused rat lungs. The effects of a calcium ionophore (CaI) were tested in isolated intact rat lungs to compare the effects of increasing intracellular Ca2+ to I/R-induced damage. Because protein kinase C (PKC) could also be a mediator of I/R injury, a PKC inhibitor was studied in lungs subjected to either I/R or CaI. In lungs subjected to I/R alone, a fivefold increase in microvascular permeability occurred after 30 min of reperfusion (P < 0.001), and a tenfold increase was present after an additional 60 min of reperfusion (P < 0.01). Pretreatment of the I/R lungs with a CaM inhibitor (trifluoperazine, 100 microM) or with a MLCK inhibitor (ML-7,500 nM) blocked the microvascular damage at both 30 and 90 min of reperfusion. When the CaM inhibitor was introduced into the venous reservoir after 46 min of reperfusion, after the microvascular damage was present, no further increase in microvascular permeability occurred. Pretreatment of the lungs with a PKC inhibitor (staurosporine, 100 nM) did not alter the magnitude of the increased microvascular permeability produced by I/R or the time course of the damage. The calcium ionophore A23187 (7.5 microM) caused increases in Kfc values similar to those produced by I/R. Pretreatment of A23187-treated lungs with a CaM inhibitor produced no protective effect on the microvascular injury at 30 min after administration. Pretreatment of the CaI-challenged lungs with staurosporine significantly increased the microvascular barrier injury at 30 min compared with that occurring with I/R. When a beta-adrenergic receptor agonist (isoproterenol, 10 microM) was introduced to the lung after CaI-induced damage had occurred, no further increase in microvascular permeability was observed, and a trend toward reversal of injury occurred. We conclude from these studies that CaM/MLCK/MLC20 system is involved in our model of I/R-induced rat lung injury but is not involved in lung injury associated with Ca2+ entering the cell.

scholarly journals Zinc exocytosis is sensitive to myosin light chain kinase inhibition in mouse and human eggs

2020 ◽  
Vol 26 (4) ◽  
pp. 228-239 ◽  
Author(s):  
Hoi Chang Lee ◽  
Maxwell E Edmonds ◽  
Francesca E Duncan ◽  
Thomas V O’Halloran ◽  
Teresa K Woodruff
Keyword(s):  
Embryo Development ◽  
Light Chain ◽  
Myosin Light Chain Kinase ◽  
Myosin Light Chain ◽  
Kinase Inhibitor ◽  
Early Embryo ◽  
Egg Activation ◽  
Cortical Granules ◽  
Preimplantation Embryo Development ◽  
Egg Maturation

Abstract Zinc dynamics are essential for oocyte meiotic maturation, egg activation, and preimplantation embryo development. During fertilisation and egg activation, the egg releases billions of zinc atoms (Zn2+) in an exocytotic event termed the ‘zinc spark’. We hypothesised that this zinc transport and exocytosis is dependent upon the intracellular trafficking of cortical granules (CG) which requires myosin-actin-dependent motors. Treatment of mature mouse and human eggs with ML-7, a myosin light chain kinase inhibitor (MLCK), resulted in an 80% reduction in zinc spark intensity compared to untreated controls when activated with ionomycin. Moreover, CG migration towards the plasma membrane was significantly decreased in ML-7-treated eggs compared with controls when activated parthenogenetically with ionomycin. In sperm-induced fertilisation via intracytoplasmic sperm injection (ICSI), ML-7-treated mouse eggs exhibited decreased labile zinc intensity and cortical CG staining. Collectively, these data demonstrate that ML-7 treatment impairs zinc release from both murine and human eggs after activation, demonstrating that zinc exocytosis requires myosin light chain kinase activity. Further, these results provide additional support that zinc is likely stored and released from CGs. These data underscore the importance of intracellular zinc trafficking as a crucial component of egg maturation necessary for egg activation and early embryo development.

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