A shear-dependent NO-cGMP-cGKI cascade in platelets acts as an auto-regulatory brake of thrombosis

Abstract

Mechanisms that limit thrombosis are poorly defined. One of the few known endogenous platelet inhibitors is nitric oxide (NO). NO activates NO sensitive guanylyl cyclase (NO-GC) in platelets, resulting in an increase of cyclic guanosine monophosphate (cGMP). Here we show, using cGMP sensor mice to study spatiotemporal dynamics of platelet cGMP, that NO-induced cGMP production in pre-activated platelets is strongly shear-dependent. We delineate a new mode of platelet-inhibitory mechanotransduction via shear-activated NO-GC followed by cGMP synthesis, activation of cGMP-dependent protein kinase I (cGKI), and suppression of Ca2+ signaling. Correlative profiling of cGMP dynamics and thrombus formation in vivo indicates that high cGMP concentrations in shear-exposed platelets at the thrombus periphery limit thrombosis, primarily through facilitation of thrombus dissolution. We propose that an increase in shear stress during thrombus growth activates the NO-cGMP-cGKI pathway, which acts as an auto-regulatory brake to prevent vessel occlusion, while preserving wound closure under low shear.

Inhibitory effects of isoscopoletin on thrombus formation via regulation of cyclic nucleotides in collagen-induced platelets

“…NO is known to increase the production of cGMP by activating NO sensitive guanylyl cyclase (NO-GC) in platelets. This effect could cause cGMP production with the increase of activated NO-GC, promoting thrombolysis [24]. In line with the results of this experiment, it was confirmed that isoscopoletin significantly increased cGMP production, possibly resulting from an increase in NO by isoscopoletin.…”

Imaging of the immune system – towards a subcellular and molecular understanding

“…It was later shown that the movement of CD3 to CD4 biosensor; a cGMP biosensor is shown as an example. cGi500 (Russwurm et al, 2007) is a cGMP indicator consisting of the engineered cGMP-binding domain of the cGMP-dependent protein kinase type I (gray) sandwiched between CFP and YFP (Wen et al, 2018). The energy transfer occurs from excited CFP to YFP.…”

“…An emerging trend is to do this using intravital imaging; this has been facilitated by recent advances in the development of improved FRET biosensors with high FRET efficiency, good specificity and excellent response kinetics (Bajar et al, 2016;Greenwald et al, 2018;Lam et al, 2012). Mice have been engineered to express FRET-based biosensors, and intravital FRET imaging has been applied to study signaling molecules, such as cGMP (Thunemann et al, 2013;Wen et al, 2018), cAMP (Sprenger et al, 2015), Ca 2+ (Yang et al, 2018), protein kinase A (PKA) and extracellular signal-regulated kinase 1/2 (ERK1/2) activity (Mizuno et al, 2014).…”

Advances, Perspectives and Potential Engineering Strategies of Light-Gated Phosphodiesterases for Optogenetic Applications

“…Sensors for cGMP were developed behind cAMP sensors because of the lower cGMP abundance in most cell types. However, several cGMP sensors are now well developed for monitoring intracellular cGMP [ 44 , 45 , 46 ].…”