G protein-coupled receptor signalling in astrocytes in health and disease: A focus on metabotropic glutamate receptors

Work published over the past 10–15 years has caused the neuroscience community to engage in a process of constant re-evaluation of the roles of glial cells in the mammalian central nervous system. Recent emerging evidence suggests that, in addition to carrying out various homeostatic functions within the CNS, astrocytes can also engage in a two-way dialogue with neurons. Astrocytes possess many of the receptors, and some of the ion channels, present in neurons endowing them with an ability to sense and respond to an array of neuronal signals. In addition, an expanding number of small molecules and proteins have been shown to be released by astrocytes in both health and disease. In this commentary we will highlight advances in our understanding of G protein-coupled receptor signalling in astrocytes, with a particular emphasis on metabotropic glutamate (mGlu) receptors. Discussion will focus on the major mGlu receptors expressed in astrocytes, mGlu3 and mGlu5, how these receptors can influence different aspects of astrocyte physiology, and how signalling by these G protein-coupled receptors might change under pathophysiological circumstances

The use of chemogenetic approaches to study the physiological roles of muscarinic acetylcholine receptors in the central nervous system

Chemical genetic has played an important role in linking specific G protein-coupled receptor (GPCR) signalling to cellular processes involved in central nervous system (CNS) functions. Key to this approach has been the modification of receptor properties such that receptors no longer respond to endogenous ligands but rather can be activated selectively by synthetic ligands. Such modified receptors have been called Receptors Activated Solely by Synthetic Ligands (RASSLs) or Designer Receptors Exclusively Activated by Designer Drugs (DREADDs). Unlike knock-out animal models which allow detection of phenotypic changes caused by loss of receptor functions, RASSL and DREADD receptors offer the possibility of rescuing "knock-out" phenotypic deficits by administration of the synthetic ligands. Here we describe the use of these modified receptors in defining the physiological role of GPCRs and validation of receptors as drug targets

Improving Physiological Relevance of Cell Culture: The Possibilities, Considerations and Future Directions of the Ex Vivo Co-Culture Model

In vitro models provide an important platform for the investigation of cellular growth and atrophy to inform, or extend mechanistic insights from, logistically challenging in vivo trials. While these models allow for the identification of candidate mechanistic pathways, many models involve supraphysiological dosages, non-physiological conditions, or experimental changes relating to individual proteins or receptors, all of which limit translation to human trials. To overcome these drawbacks, the use of ex vivo human plasma and serum has been used in cellular models to investigate changes in myotube hypertrophy, cellular protein synthesis, anabolic and catabolic markers in response to differing age, disease states, and nutrient status. However, there are currently no concurrent guidelines outlining the optimal methodology for this model. This review discusses the key methodological considerations surrounding the use of ex vivoplasma and serum, with a focus in application to skeletal muscle cell lines (i.e., C2C12, L6 and LHCN-M2) and human primary skeletal muscle cells (HSMC) as a means to investigate molecular signaling in models of atrophy and hypertrophy, alongside future directions

Sensitivity of palaeotidal models of the northwest European shelf seas to glacial isostatic adjustment since the Last Glacial Maximum

AbstractThe spatial and temporal distribution of relative sea-level change over the northwest European shelf seas has varied considerably since the Last Glacial Maximum, due to eustatic sea-level rise and a complex isostatic response to deglaciation of both near- and far-field ice sheets. Because of the complex pattern of relative sea level changes, the region is an ideal focus for modelling the impact of significant sea-level change on shelf sea tidal dynamics. Changes in tidal dynamics influence tidal range, the location of tidal mixing fronts, dissipation of tidal energy, shelf sea biogeochemistry and sediment transport pathways. Significant advancements in glacial isostatic adjustment (GIA) modelling of the region have been made in recent years, and earlier palaeotidal models of the northwest European shelf seas were developed using output from less well-constrained GIA models as input to generate palaeobathymetric grids. We use the most up-to-date and well-constrained GIA model for the region as palaeotopographic input for a new high resolution, three-dimensional tidal model (ROMS) of the northwest European shelf seas. With focus on model output for 1 ka time slices from the Last Glacial Maximum (taken as being 21 ka BP) to present day, we demonstrate that spatial and temporal changes in simulated tidal dynamics are very sensitive to relative sea-level distribution. The new high resolution palaeotidal model is considered a significant improvement on previous depth-averaged palaeotidal models, in particular where the outputs are to be used in sediment transport studies, where consideration of the near-bed stress is critical, and for constraining sea level index points

The role of megatides and relative sea level in controlling the deglaciation of the British-Irish and Fennoscandinavian ice sheets

This is the final version of the article. Available from Wiley via the DOI in this record.Key external forcing factors have been proposed to explain the collapse of ice sheets, including atmospheric and ocean temperatures, subglacial topography, relative sea level and tidal amplitudes. For past ice sheets it has not hitherto been possible to separate relative sea level and tidal amplitudes from the other controls to analyse their influence on deglaciation style and rate. Here we isolate the relative sea level and tidal amplitude controls on key ice stream sectors of the last British–Irish and Fennoscandian ice sheets using published glacial isostatic adjustment models, combined with a new and previously published palaeotidal models for the NE Atlantic since the Last Glacial Maximum (22 ka BP). Relative sea level and tidal amplitude data are combined into a sea surface elevation index for each ice stream sector demonstrating that these controls were potentially important drivers of deglaciation in the western British Irish Ice Sheet ice stream sectors. In contrast, the Norwegian Channel Ice Stream was characterized by falling relative sea level and small tidal amplitudes during most of the deglaciation. As these simulations provide a basis for observational field testing we propose a means of identifying the significance of sea level and tidal amplitudes in ice sheet collapse.Funding was provided by the Natural Environment Research Council (NERC) through grant NE/I527853/1 (PhD studentship to S.L.W.). The research was supported by the Climate Change Consortium of Wales and the NERC BRITICE-CHRONO Consortium grant (NE/J007579/1). Jess Vaughan and Martyn Roberts drafted Figs 2–5

An interactive visualization and data portal tool (PALTIDE) for relative sea level and palaeotidal simulations of the northwest European shelf seas since the Last Glacial Maximum

ABSTRACTRelative sea level (RSL) predictions based on glacial isostatic adjustment (GIA) simulations and palaeotidal predictions generated by hydrodynamic models using GIA‐generated palaeotopographies are available in the published literature, and datasets are available via data repositories. However, these data are often difficult to extract for specific locations or timeslices, requiring users to request datasets from corresponding authors. To overcome the intractability of these data and to enable users to interrogate datasets themselves without requiring offline requests, we have developed PALTIDE, an online visualization tool with intuitive user interface accessible at https://shiny.bangor.ac.uk/paleotidal/. The model domain for this interactive visualization tool is the northwest European continental shelf, covering the period from the Last Glacial Maximum (LGM) to the present day, and is based on previous GIA simulations by Bradley and colleagues and hydrodynamic simulations using Regional Ocean Modelling System (ROMS) published by Ward and colleagues. The tool is developed in R and utilizes a number of packages including shiny and bslib for the frontend, and arrow, raster and the tidyverse for backend data processing. The tool enables visualizations and data downloads for RSL, tidal amplitude and tide‐dependent parameters for any location within the model domain over 1000‐year timesteps from the LGM to the present

Preclinical efficacy of hK2 targeted [177Lu]hu11B6 for prostate cancer theranostics

Androgen ablating drugs increase life expectancy in men with metastatic prostate cancer, but resistance inevitably develops. In a majority of these recurrent tumors, the androgen axis is reactivated in the form of increased androgen receptor (AR) expression. Targeting proteins that are expressed as a down-stream effect of AR activity is a promising rationale for management of this disease. The humanized IgG1 antibody hu11B6 internalizes into prostate and prostate cancer (PCa) cells by binding to the catalytic cleft of human kallikrein 2 (hK2), a prostate specific enzyme governed by the AR-pathway. In a previous study, hu11B6 conjugated with Actinium-225 (225Ac), a high linear energy transfer (LET) radionuclide, was shown to generate an AR-upregulation driven feed-forward mechanism that is believed to enhance therapeutic efficacy. We assessed the efficacy of hu11B6 labeled with a low LET beta-emitter, Lutetium-177 (177Lu) and investigated whether similar tumor killing and AR-enhancement is produced. Moreover, single-photon emission computed tomography (SPECT) imaging of 177Lu is quantitatively accurate and can be used to perform treatment planning. [177Lu]hu11B6 therefore has significant potential as a theranostic agent. Materials and Methods: Subcutaneous PCa xenografts (LNCaP s.c.) were grown in male mice. Biokinetics at 4-336 h post injection and uptake as a function of the amount of hu11B6 injected at 72 h were studied. Over a 30 to 120-day treatment period the therapeutic efficacy of different activities of [177Lu]hu11B6 were assessed by volumetric tumor measurements, blood cell counts, molecular analysis of the tumor as well as SPECT/CT imaging. Organ specific mean absorbed doses were calculated, using a MIRD-scheme, based on biokinetic data and rodent specific S-factors from a modified MOBY phantom. Tumor tissues of treated xenografts were immunohistochemically (IHC) stained for Ki-67 (proliferation) and AR, SA-β-gal activity (senescence) and analyzed by digital autoradiography (DAR). Results: Organ-to-blood and tumor-to-blood ratios were independent of hu11B6 specific activity except for the highest amount of antibody (150 µg). Tumor accumulation of [177Lu]hu11B6 peaked at 168 h with a specific uptake of 29 ± 9.1 percent injected activity per gram (%IA/g) and low accumulation in normal organs except in the submandibular gland (15 ± 4.5 %IA/g), attributed to a cross-reaction with mice kallikreins in this organ, was seen. However, SPECT imaging with therapeutic amounts of [177Lu]hu11B6 revealed no peak in tumor accumulation at 7 d, probably due to cellular retention of 177Lu and decreasing tumor volumes. For [177Lu]hu11B6 treated mice, tumor decrements of up to 4/5 of the initial tumor volume and reversible myelotoxicity with a nadir at 12 d were observed after a single injection. Tumor volume reduction correlated with injected activity and the absorbed dose. IHC revealed retained expression of AR throughout treatment and that Ki-67 staining reached a nadir at 9-14 d which coincided with high SA- β-gal activity (14 d). Quantification of nuclei staining showed that Ki-67 expression correlated negatively with activity uptake. AR expression levels in cells surviving therapy compared to previous timepoints and to controls at 30 d were significantly increased (p = 0.017). Conclusions: This study shows that hu11B6 labeled with the low LET beta-emitting radionuclide 177Lu can deliver therapeutic absorbed doses to prostate cancer xenografts with transient hematological side-effects. The tumor response correlated with the absorbed dose both on a macro and a small scale dosimetric level. Analysis of AR staining showed that AR protein levels increased late in the study suggesting a therapeutic mechanism, a feed forward mechanism coupled to AR driven response to DNA damage or clonal lineage selection, similar to that reported in high LET alpha-particle therapy using 225Ac labeled hu11B6, however emerging at a later timepoint

Borders and Catastrophe: lessons from COVID-19 for the European Green Deal

This article considers how the European Union and Member States’ responses to the COVID-19 crisis in the first half of 2020 could inform climate action in Europe, and particularly the resumption of actions on the EGD. It first outlines the EU’s public health and economic responses to COVID-19 and Europe’s role in the global response to the pandemic. We find that, based on the challenges and successes of all these responses, a strong argument can be made for ‘more Europe’ – greater integration, and stronger EU-level institutions – to lead and govern the COVID-19 response. This has direct lessons for the governance and scope of future climate action

Editorial for Advances in G Protein-Coupled Receptor Signal Transduction Special Issue

No abstract available