From Mechanical Force to RhoA Activation

Throughout their lives all cells constantly experience and respond to various mechanical forces. These frequently originate externally but can also arise internally as a result of the contractile actin cytoskeleton. Mechanical forces trigger multiple signaling pathways. Several converge and result in the activation of the GTPase RhoA. In this review we focus on the pathways by which mechanical force leads to RhoA regulation, especially when force is transmitted via cell adhesion molecules that mediate either cell-matrix or cell-cell interactions. We discuss both the upstream signaling events that lead to activation of RhoA, as well as the downstream consequences of this pathway. These include not only cytoskeletal reorganization and, in a positive feedback loop, increased myosin-generated contraction, but also profound effects on gene expression and differentiation

AMPK alpha 1-induced RhoA phosphorylation mediates vasoprotective effect of estradiol

OBJECTIVE: Estradiol (E2) mediates numerous beneficial effects assigned to estrogens, but whereas mechanisms have been described at the endothelial level, direct effects on vascular smooth muscle cells (VSMC) are poorly documented. As evidence accumulates regarding the role of RhoA in vascular pathophysiology and the benefit of RhoA-Rho associated protein kinase (Rock) pathway inhibition, we analyzed if E2 could inhibit it in VSMC. METHODS AND RESULTS: We show that in VSMC, E2 inhibits the RhoA-Rock pathway in a time- and concentration-dependent manner. The inhibition of RhoA-Rock pathway results from E2-induced phosphorylation of the Ser188 of RhoA. Using pharmacological, transfection, and in vitro phosphorylation experiments, we demonstrate that AMP-activated protein kinase subunit alpha 1 (AMPKalpha1) is activated by estrogen receptor stimulation and catalyzes RhoA phosphorylation induced by E2. Ex vivo, ovariectomy leads to an increase in the amplitude of phenylephrine- or serotonine-induced contractions of aortic rings in wild-type mice but not in AMPKalpha1-knock-out mice or E2-supplemented animals. These functional effects were correlated with a reduced level of RhoA phosphorylation in the aorta of ovariectomized female, male, and AMPKalpha1 knock-out mice. CONCLUSION: Our work thus defines AMPKalpha1 as (1) a new kinase for RhoA and (2) a new mediator of the vasoprotective effects of estrogen

Two images of Nantes as a ‘Green Model’ of Urban Planning and Governance: The ‘Collaborative City’ Versus the ‘Slow City’

This article examines how the city of Nantes, European Green Capital in 2013, came to promote plans for a new international airport at Notre-Dame-des-Landes. Deploying poststructuralist discourse theory, it analyses how the highly politicised struggle against the airport reveals the limits of the Nantes model of urban sustainability and collaboration, giving rise to a counter model, which we provisionally characterise as the ‘slow city’. While the struggle against the airport can be understood as a rural social movement, we show how its ideals and logics have been progressively displaced to Nantes itself, disclosing new images and possibilities of urban governance

Contextual effects of immigrant presence on populist radical right support: testing the ‘halo effect’ on Front National voting in France

This paper examines the relationship between immigration and populist radical right (PRR) support, based on an analysis of the contextual effects of immigrant presence on Front National vote in France in 2017. Using a unique set of survey data geolocalising respondents at the subcommunal level, it finds evidence for the existence of a curvilinear “halo effect,” with substantial increases in the probability of PRR vote in areas surrounding communities with significantly higher-than-average immigrant populations, and independent of other socio-economic context, as well as individual socio-demographic characteristics. Most importantly, a path analysis confirms the presence of individual attitudinal mediators of this halo effect on PRR vote, thus testing the foundation of the halo, namely that the contextual effects of immigrant presence act on attitudes which drive PRR support. These findings provide a significant step forward in understanding the mechanisms linking subjective experience of immigration with voting for the populist radical right

The RhoA Guanine Nucleotide Exchange Factor, LARG, Mediates ICAM-1-Dependent Mechanotransduction in Endothelial Cells To Stimulate Transendothelial Migration

RhoA-mediated cytoskeletal rearrangements in endothelial cells (ECs) play an active role in leukocyte transendothelial cell migration (TEM), a normal physiological process in which leukocytes cross the endothelium to enter the underlying tissue. Although much has been learned about RhoA signaling pathways downstream from ICAM-1 in ECs, little is known about the consequences of the tractional forces that leukocytes generate on ECs as they migrate over the surface before TEM. We have found that after applying mechanical forces to ICAM-1 clusters, there is an increase in cellular stiffening and enhanced RhoA signaling compared with ICAM-1 clustering alone. We have identified that leukemia-associated Rho guanine nucleotide exchange factor (LARG), also known as Rho GEF 12 (ARHGEF12) acts downstream of clustered ICAM-1 to increase RhoA activity, and that this pathway is further enhanced by mechanical force on ICAM-1. Depletion of LARG decreases leukocyte crawling and inhibits TEM. To our knowledge, this is the first report of endothelial LARG regulating leukocyte behavior and EC stiffening in response to tractional forces generated by leukocytes

The GAPS programme at TNG XXII. The GIARPS view of the extended helium atmosphere of HD189733 b accounting for stellar activity

Exoplanets orbiting very close to their host star are strongly irradiated. This can lead the upper atmospheric layers to expand and evaporate into space. The metastable helium (HeI) triplet at 1083.3nm has recently been shown to be a powerful diagnostic to probe extended and escaping exoplanetary atmosphere. We perform high-resolution transmission spectroscopy of the transiting hot Jupiter HD189733b with the GIARPS (GIANO-B + HARPS-N) observing mode of the Telescopio Nazionale Galileo, taking advantage of the simultaneous optical+near infrared spectral coverage to detect HeI in the planet's extended atmosphere and to gauge the impact of stellar magnetic activity on the planetary absorption signal. Observations were performed during five transit events of HD189733b. By comparison of the in- and out-of-transit GIANO-B observations we compute high-resolution transmission spectra, on which we perform equivalent width measurements and light-curves analyses to gauge the excess in-transit absorption in the HeI triplet. We detect an absorption signal during all five transits. The mean in-transit absorption depth amounts to 0.75+/-0.03%. We detect night-to-night variations in the HeI absorption signal likely due to the transit events occurring in presence of stellar surface inhomogeneities. We evaluate the impact of stellar-activity pseudo-signals on the true planetary absorption using a comparative analysis of the HeI and the H$\alpha$ lines. We interpret the time-series of the HeI absorption lines in the three nights not affected by stellar contamination -exhibiting a mean in-transit absorption depth of 0.77+/-0.04%- using a 3-d atmospheric code. Our simulations suggest that the helium layers only fill part of the Roche lobe. Observations can be explained with a thermosphere heated to $\sim$12000 K, expanding up to $\sim$1.2 planetary radii, and losing $\sim$1 g/s of metastable helium.Comment: 17 pages, 17 figures, accepted for publication in A&

The Rho exchange factor Arhgef1 mediates the effects of angiotensin II on vascular tone and blood pressure

Hypertension is one of the most frequent pathologies in the industrialized world. Although recognized to be dependent on a combination of genetic and environmental factors, its molecular basis remains elusive. Increased activity of the monomeric G protein RhoA in arteries is a common feature of hypertension. However, how RhoA is activated and whether it has a causative role in hypertension remains unclear. Here we provide evidence that Arhgef1 is the RhoA guanine exchange factor specifically responsible for angiotensin II-induced activation of RhoA signaling in arterial smooth muscle cells. We found that angiotensin II activates Arhgef1 through a previously undescribed mechanism in which Jak2 phosphorylates Tyr738 of Arhgef1. Arhgef1 inactivation in smooth muscle induced resistance to angiotensin II-dependent hypertension in mice, but did not affect normal blood pressure regulation. Our results show that control of RhoA signaling through Arhgef1 is central to the development of angiotensin II-dependent hypertension and identify Arhgef1 as a potential target for the treatment of hypertension

Altered Gene Expression in Pulmonary Tissue of Tryptophan Hydroxylase-1 Knockout Mice: Implications for Pulmonary Arterial Hypertension

The use of fenfluramines can increase the risk of developing pulmonary arterial hypertension (PAH) in humans, but the mechanisms responsible are unresolved. A recent study reported that female mice lacking the gene for tryptophan hydroxylase-1 (Tph1(−/−) mice) were protected from PAH caused by chronic dexfenfluramine, suggesting a pivotal role for peripheral serotonin (5-HT) in the disease process. Here we tested two alternative hypotheses which might explain the lack of dexfenfluramine-induced PAH in Tph1(−/−) mice. We postulated that: 1) Tph1(−/−) mice express lower levels of pulmonary 5-HT transporter (SERT) when compared to wild-type controls, and 2) Tph1(−/−) mice display adaptive changes in the expression of non-serotonergic pulmonary genes which are implicated in PAH. SERT was measured using radioligand binding methods, whereas gene expression was measured using microarrays followed by quantitative real time PCR (qRT-PCR). Contrary to our first hypothesis, the number of pulmonary SERT sites was modestly up-regulated in female Tph1(−/−) mice. The expression of 51 distinct genes was significantly altered in the lungs of female Tph1(−/−) mice. Consistent with our second hypothesis, qRT-PCR confirmed that at least three genes implicated in the pathogenesis of PAH were markedly up-regulated: Has2, Hapln3 and Retlna. The finding that female Tph1(−/−) mice are protected from dexfenfluramine-induced PAH could be related to compensatory changes in pulmonary gene expression, in addition to reductions in peripheral 5-HT. These observations emphasize the intrinsic limitation of interpreting data from studies conducted in transgenic mice that are not fully characterized

The GAPS programme at TNG. XLV. HI Balmer lines transmission spectroscopy and NLTE atmospheric modelling of the ultra-hot Jupiter KELT-20b/MASCARA-2b

We aim at extracting the transmission spectrum of the HI Balmer lines of the ultra-hot Jupiter (UHJ) KELT-20b/MASCARA-2b from observations and to further compare the results with what obtained through forward modelling accounting for non-local thermodynamic equilibrium (NLTE) effects. We extract the line profiles from six transits obtained with the HARPS-N high-resolution spectrograph attached to the Telescopio Nazionale Galileo telescope. We compute the temperature-pressure (TP) profile employing the helios code in the lower atmosphere and the Cloudy NLTE code in the middle and upper atmosphere. We further use Cloudy to compute the theoretical planetary transmission spectrum in LTE and NLTE for comparison with observations. We detected the Halpha (0.79+/-0.03%; 1.25 Rp), Hbeta (0.52+/-0.03%; 1.17 Rp), and Hgamma (0.39+/-0.06%; 1.13 Rp) lines, while we detected the Hdelta line at almost 4 sigma (0.27+/-0.07%; 1.09 Rp). The models predict an isothermal temperature of about2200 K at pressures >10^-2 bar and of about 7700 K at pressures <10^-8 bar, with a roughly linear temperature rise in between. In the middle and upper atmosphere, the NLTE TP profile is up to about 3000 K hotter than in LTE. The synthetic transmission spectrum derived from the NLTE TP profile is in good agreement with the observed HI Balmer line profiles, validating our obtained atmospheric structure. Instead, the synthetic transmission spectrum derived from the LTE TP profile leads to significantly weaker absorption compared to the observations. Metals appear to be the primary agents leading to the temperature inversion in UHJs and the impact of NLTE effects on them increases the magnitude of the inversion. We find that the impact of NLTE effects on the TP profile of KELT-20b/MASCARA-2b is larger than for the hotter UHJ KELT-9b, and thus NLTE effects might be relevant also for planets cooler than KELT-20b/MASCARA-2b.Comment: Accepted for publication in Astronomy and Astrophysic