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Balmes, A.

Publications and source records attributed to Balmes, A..

3 recordsLinked to original sources

Distinct nanoscale dynamics of growth receptor complexes link hormone perception to rapid cell wall remodeling

Cell elongation is a fundamental process allowing plants to change size and shape, a process governed by several growth promoting hormones. While hormones such as brassinosteroids (BRs) and phytosulfokines (PSKs) have been shown to play important roles in elongation growth, the early steps of signal perception remains elusive, especially with regard to PSK. Here we report a rapid mechanism by which PSK alter cell wall mechanical properties in elongating hypocotyls. Notably, this mode of action differs from that of BR. Making use of atomic force microscopy and fluorescence lifetime imaging microscopy we demonstrate how hallmarks of growing plant cells such as mechanical wall properties, porosity and apoplastic pH are differentially affected by PSK compared to BR. Using super-resolution microscopy, we show that the receptor complex components for BR and PSK display individual spatiotemporal movement and organization patterns. The BR receptor BRI1 transitions to a faster diffusive state upon ligand perception, while the PSK receptor PSKR1 associates in tighter clusters. The shared co-receptor BAK1 displays a selective decrease in cluster density after PSK treatment. We found that the putative cell wall state sensor RLP44 is required for the observed changes, however the spatiotemporal dynamics of RLP44 are not altered during signaling. We propose a model of how cell walls are specifically tuned by BRI1- and PSKR1-centered signaling hubs as potential prerequisites for and during cell elongation initiation.

plant biology↗

Pharmacological Activation of NO-Sensitive Guanylyl Cyclase Ameliorates Obesity-Induced Arterial Stiffness

ObjectiveArterial stiffness, or loss of elastic compliance in large arteries, is an independent precursor of cardiovascular disease (CVD)1 and dementia2. Akin to anti-hypertensive and lipid-lowering drugs, arterial de-stiffening therapies could be beneficial at decreasing CVD risk. We previously discovered that enhanced cytoskeletal actin polymerization in vascular smooth muscle cells (VSMCs) contributes to increased arterial stiffness3. In aortas and VSMCs, we previously found that decreased NO-sensitive guanylyl cyclase (NO-GC), the NO receptor which synthesizes cGMP, caused downregulation of cGMP-dependent protein kinase I (cGKI) and of its target vasodilator-stimulated phosphoprotein (pVASPS239), leading to increased cytoskeletal actin polymerization3. In the current study, we tested whether activating NO-GC with an NO-GC activator (cinaciguat) modulates pVASPS239 and cytoskeletal actin polymerization in VSMCs, thereby preventing obesity-induced arterial stiffness. Approach & ResultsCinaciguat administration (5 mg/kg) to high fat, high sucrose diet (HFHS)-fed mice, our established model of arterial stiffness4, (1) decreased pulse wave velocity, the in vivo index of arterial stiffness, without affecting blood pressure, (2) increased aortic pVASPS239 levels, and (3) decreased the ratio of filamentous (F) to globular (G) actin, compared to vehicle administration. In cultured VSMCs, cinaciguat (10 mol/L) increased pVASPS239 levels and decreased the F/G actin ratio at baseline and after stimulation with the cytokine tumor necrosis factor (TNF), used to mimic the inflammatory milieu of HFHS aortas. These effects were abrogated in aortas and VSMCs from mice with smooth muscle-specific cGKI deletion (cGKISMKO), while being mimicked by a cell-permeable cGMP analog (8-Br-cGMP, 1 mol/L), which also decreased VSMC stiffness in vitro. ConclusionsCollectively, our data strongly support the notion that pharmacological NO-GC activation would be beneficial in decreasing obesity-associated arterial stiffness by decreasing VSMC cytoskeletal actin hyper-polymerization. If translated to humans, NO-GC activators could become a viable approach to clinically treat arterial stiffness, which remains an unmet medical need.

pharmacology and toxicology↗

Role of the NO-GC/cGMP signaling pathway in platelet biomechanics

Cyclic guanosine monophosphate (cGMP) is a second messenger produced by the NO-sensitive guanylyl cyclase (NO-GC) enzyme. In platelets, the NO-GC/cGMP pathway inhibits aggregation. One aspect of the inhibitory mechanism involves changes in the cytoskeleton; however, the molecular mechanisms underlying platelet inhibition and its correlation with cytoskeletal cellular stiffness are poorly understood. We measured the cellular stiffness of individual platelets after treatment with the NO-GC stimulator riociguat or the NO-GC activator cinaciguat, using scanning ion conductance microscopy (SICM). We quantified changes in platelet shape using deep learning-based platelet morphometry. Cytoskeletal actin polymerization and platelet activation were measured by co-immunostaining F-actin and P-selectin, respectively. To test for clinical applicability of NO-GC stimulators in the context of increased thrombogenicity risk, we investigated the effect of riociguat on platelets from human immunodeficiency virus (HIV)-positive patients taking abacavir sulphate (ABC)-containing regimens, compared with HIV-negative volunteers. Stimulation of human and murine platelets with the NO-GC stimulator riociguat or with the NO-GC activator cinaciguat downregulated P-selectin expression, decreased F-actin polymerization, and decreased cellular stiffness by {approx}50%, compared to vehicle control. In addition, platelets became more circular, indicating decreased activation. Riociguat did not cause any change in platelet aggregation or circularity in HIV-positive patients taking ABC-containing regimens. These results corroborate a functional role of the NO-GC enzyme in platelet biomechanics (cellular stiffness) in correlation with the inhibition of platelet activation and morphological changes. The observed changes in stiffness and platelet shape therefore demonstrate the possibility of pharmacologically targeting the NO-GC/cGMP pathway.

biochemistry↗