Thrombin-Induced Endothelin-1 Synthesis and Secretion in Retinal Pigment Epithelial Cells Is Rho Kinase Dependent

Narayan, Santosh; Prasanna, Ganesh; Tchedre, Kissaou; Krishnamoorthy, Raghu R; Yorio, Thomas

doi:10.1089/jop.2010.0072

Cited by 8 publications

(6 citation statements)

References 54 publications

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“…We identified PAR1 as the main receptor responsible for thrombin-induced [Ca 2+ ]i increase, since PAR1-AP elicited [Ca 2+ ]i increase equivalent to that induced by thrombin ( Figure 2 ), whereas stimulation by PAR3-AP or PAR4-AP did not. These results are in agreement with findings in epithelial cells [ 44 , 45 ], although in contrast with a previous report showing the increase in [Ca 2+ ]i induced by PAR4 stimulation in ARPE-19 cells [ 33 ]. This discrepancy could be ascribed to the use by Narayan et al (2010) of a synthetic PAR4-AP 10 times more potent than the endogenous agonist peptide used in the present work [ 46 ].…”

Section: Discussionsupporting

confidence: 92%

“…The effect of thrombin stimulation on RPE cell [Ca +2 ]i was determined using fluorescence microscopy, as described in the Methods. Although thrombin-induced increase in [Ca +2 ]i in RPE cells has been reported [ 32 , 33 ], the characteristics of this response as a function of the intensity and duration of the stimulus, determinant for functional outcome, have not been analyzed.…”

Section: Resultsmentioning

confidence: 99%

“…Thrombin-induced [Ca 2+ ]i mobilization in RPE cells has been reported [ 32 , 33 ]. Since calpain is a Ca 2+ activated protease, we showed that thrombin induces the specific, dose-dependent increase in [Ca 2+ ]i with an Ec 50 of 0.55 nM ( Figure 1(c) ).…”

Section: Discussionmentioning

confidence: 99%

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Thrombin-Induced Calpain Activation Promotes Protease-Activated Receptor 1 Internalization

Alvarez-Arce

Lee-Rivera

López

et al. 2017

International Journal of Cell Biology

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The serine protease thrombin activates Protease-Activated Receptors (PARs), a family of G-protein-coupled receptors (GPCRs) activated by the proteolytic cleavage of their extracellular N-terminal domain. Four members of this family have been identified: PAR1-4. The activation of Protease-Activated Receptor 1(PAR1), the prototype of this receptor family, leads to an increase in intracellular Ca +2 concentration ([Ca +2 ]i) mediated by G q11 coupling and phospholipase C (PLC) activation. We have previously shown that the stimulation of PAR1 by thrombin promotes intracellular signaling leading to RPE cell transformation, proliferation, and migration which characterize fibroproliferative eye diseases leading to blindness. Within this context, the elucidation of the mechanisms involved in PAR1 inactivation is of utmost importance. Due to the irreversible nature of PAR1 activation, its inactivation must be efficiently regulated in order to terminate signaling. Using ARPE-19 human RPE cell line, we characterized thrombininduced [Ca +2 ]i increase and demonstrated the calcium-dependent activation of -calpain mediated by PAR1. Calpains are a family of calcium-activated cysteine proteases involved in multiple cellular processes including the internalization of membrane proteins through clathrin-coated vesicles. We demonstrated that PAR1-induced calpain activation results in the degradation of -spectrin by calpain, essential for receptor endocytosis, and the consequent decrease in PAR1 membrane expression. Collectively, the present results identify a novel -calpain-dependent mechanism for PAR1 inactivation following exposure to thrombin.

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Section: Discussionsupporting

confidence: 92%

Section: Resultsmentioning

confidence: 99%

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Thrombin-Induced Calpain Activation Promotes Protease-Activated Receptor 1 Internalization

Alvarez-Arce

Lee-Rivera

López

et al. 2017

International Journal of Cell Biology

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“…These finding indicate that OFQ activates RhoA directly via its own NOP receptor rather than it being an indirect effect through another receptor or pathway. The activation of RhoA by OFQ is transient, which is consistent with other studies showing transient activation of RhoA following treatments with molecules such as tumor necrosis factor (Hunter et al, 2003), thrombin (Narayan et al, 2010), vascular endothelial growth factor (Sun et al, 2006;Yokomori et al, 2009), and carbachol (Liu et al, 2006). Acute activation of RhoA may induce downstream signaling pathways that then have effects on dendrite outgrowth.…”

Section: Discussionsupporting

confidence: 89%

Neuropeptide orphanin FQ inhibits dendritic morphogenesis through activation of RhoA

Alder

Kallman

Palmieri

et al. 2013

Developmental Neurobiology

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Brain-derived neurotrophic factor (BDNF) plays a facilitatory role in neuronal development and promotion of differentiation. Mechanisms that oppose BDNF's stimulatory effects create balance and regulate dendritic growth. However, these mechanisms have not been studied. We have focused our studies on the BDNF-induced neuropeptide OrphaninFQ/ Nociceptin (OFQ); while BDNF is known to enhance synaptic activity, OFQ has opposite effects on activity, learning, and memory. We have now examined whether OFQ provides a balance to the stimulatory effects of BDNF on neuronal differentiation in the hippocampus. Golgi staining in OFQ knockout (KO) mice revealed an increase in primary dendrite length as well as spine density, suggesting that endogenous OFQ inhibits dendritic morphology. We have also used cultured hippocampal neurons to demonstrate that exogenous OFQ has an inhibitory effect on dendritic growth and that the neuropeptide alters the response to BDNF when pre-administered. To determine if BDNF and OFQ act in a feedback loop, we inhibited the actions of the BDNF and OFQ receptors, TrkB and NOP using ANA-12 and NOP KO mice respectively but our data suggest that the two factors do not act in a negative feedback loop. We found that the inhibition of dendritic morphology induced by OFQ is via enhanced RhoA activity. Finally, we have evidence that RhoA activation is required for the inhibitory effects of OFQ on dendritic morphology. Our results reveal basic mechanisms by which neurons not only regulate the formation of proper dendritic growth during development but also control plasticity in the mature nervous system.

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“…Importantly, the RPE is a major source of endothelin-1 at the outer blood-retinal barrier, a potent vasoactive pep-tide. There is consistent evidence that PAR-1 regulates endothelin synthesis not by calcium mobilisation but rather by rho/Rho kinase activation (30).…”

Section: Retinamentioning

confidence: 91%

Vorapaxar and diplopia: Possible off-target PAR-receptor mismodulation

Serebruany

Fortmann²,

Rao

et al. 2016

Thromb Haemost

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SummaryVorapaxar, a novel antiplatelet thrombin PAR-1 inhibitor, has been evaluated in the successful TRA2P trial and the failed TRACER trial. The drug is currently approved for post myocardial infarction and peripheral artery disease indications with concomitant use of clopidogrel and/or aspirin. The FDA ruled that the vorapaxar safety profile is acceptable. However, both trials revealed excess diplopia (double vision) usually reversible after vorapaxar. The diplopia risk appears to be small (about 1 extra case per 1,000 treated subjects), but real. Overall, there were 10 placebo and 34 vorapaxar diplopia cases (p=0.018) consistent for TRACER (2 vs 13 cases; p=0.010) and for TRA2P (8 vs 21 cases; p=0.018). Hence, we review the FDA-confirmed evidence and discuss potential causes and implications of such a surprising adverse association, which may be related to off-target PAR receptor mismodulation in the eye.

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Thrombin-Induced Endothelin-1 Synthesis and Secretion in Retinal Pigment Epithelial Cells Is Rho Kinase Dependent

Abstract: Transient intracellular calcium mobilization and protein kinase C activation by thrombin play a minor role, if any, in ET-1 synthesis in RPE. Instead, rho/Rho kinase activation after PAR-1 stimulation strongly increased ppET-1 mRNA and ET-1 secretion in hRPE cells.

Cited by 8 publications

References 54 publications

Thrombin-Induced Calpain Activation Promotes Protease-Activated Receptor 1 Internalization

Thrombin-Induced Calpain Activation Promotes Protease-Activated Receptor 1 Internalization

Neuropeptide orphanin FQ inhibits dendritic morphogenesis through activation of RhoA

Vorapaxar and diplopia: Possible off-target PAR-receptor mismodulation

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